ПРИМЕНЕНИЕ КУЛЬТУРАЛЬНОЙ ЖИДКОСТИ, РАЗЖИЖАЮЩЕЙ НЕФТЬ И НЕФТЕПРОДУКТЫ РАЗЛИЧНОЙ ВЯЗКОСТИ. СПОСОБ ПОЛУЧЕНИЯ КУЛЬТУРАЛЬНОЙ ЖИДКОСТИ, РАЗЖИЖИВАЮЩЕЙ НЕФТЬ И НЕФТЕПРОДУКТЫ РАЗЛИЧНОЙ ВЯЗКОСТИ

... 1. Применение культуральной жидкости в качестве разжижающего вещества нефти в пластах, трубопроводах, емкостях, для создания водно-мазутного топлива. Также предлагаемая культуральная жидкость может использоваться для получения из нее жидких углеводородов. При очистке емкостей от углеводородсодержащего осадка, культуральная жидкость как очищающее средство перемешивается с осадком и обеспечивает возможность откачать образовавшуюся смесь-пульпу, отличается тем, что эмульсионные свойства культуральной жидкости обеспечиваются наличием углеводородов в культуральной жидкости в количестве не менее 10 г/л и рассредоточены в жидкости происхождением на молекулярном уровне. 2. Применение по п. 1 отличается тем, что как разжижающее средство, культуральная жидкость образует эмульсию с нефтью и нефтепродуктами до любой степени вязкости, от вязкости воды до вязкости самого нефтепродукта. 3. Способ получения культуральной жидкости отличается тем, что процесс осуществляется в анаэробных условиях в присутствии ...

СПОСОБ ПОЛУЧЕНИЯ БИОТОПЛИВА

Способ получения биотоплива из органического сырья, включающий обработку измельченного исходного сырья микроорганизмами в присутствии соединений, являющимися донорами водорода, отличающийся тем, что в качестве основного сырья используют растение эйхорнию и ее смеси с торфом, лигнином и со сбраженными навозной жижей, содержимым желудка животных и активным илом аэротенка, которые включают метаногенные микроорганизмы с культуральной средой на основе фосфатно-хлоридно-карбонатного буфера, а также катализаторы интенсификации процесса выделения биогаза в виде отходов переработки древесины, содержащих гуматы, целлюлозу и/или отходы сахарного производства, причем процесс ведут в начале при температуре от 15 до 70°С в анаэробных или аэробных условиях, отбирают синтезированный метан, а образующийся диоксид углерода оставляют в реакторе, добавляют вещества-доноры водорода, гуминовые соединения, микроорганизмы, содержимое реактора перемешивают и процесс далее ведут при температуре от 3 до 60°C и рН ...

Метантенк

Способ получения @ -каротина из суспензии водорослей рода DUNaLIeLLa в растворе хлорида натрия с концентрацией не менее 3 М

Изобретение относится к биотехнологии и касается получения витаминов. Целью изобретения является улучшение качества β-каротина и упрощение процесса. Это достигается тем, что водоросли рода DUNALIELLA в растворе хлорида натрия концентрацией не менее 3М контактируют с гидрофобным корпускулярным или волокнистым адсорбентом, обрабатывают водоросли растворителем для экстракции β-каротина, свободного от нежелательных каротиноидов и липидов, а затем отгоняют растворитель из экстракта.

MONOCLONAL ANTIBODY DIRECTED TO HUMAN GANGLIOSIDE GD 2?

MONOCLONAL ANTIBODIES

Process for the selective production of hydrocarbon based fuels from algae utilizing water at subcritical conditions

PROCESSES FOR BIOCONVERSION OF CARBON BEARING MATERIALS

Processing biomass.

Method and bioreactor for producing synfuel from carbonaceous material

PROCESSES FOR BIOCONVERSION OF CARBON BEARING MATERIALS

Algae growth for biofuels

Method and bioreactor for producing synfuel from carbonaceous material

PROCESSES FOR BIOCONVERSION OF CARBON BEARING MATERIALS

Process and apparatus for extracting biodiesel from algae.

Processing biomass.

Algae growth for biofuels.

Process for the selective production of hydrocarbon based fuels from algae utilizing water at subcritical conditions

Transgenic amorpha-4, 11-diene synthesis.

Process for the selective production of hydrocarbon based fuels from algae utilizing water at subcritical conditions

Process and apparatus for extracting biodiesel from algae.

Processing biomass.

Algae growth for biofuels

Algae growth for biofuels

Method and bioreactor for producing synfuel from carbonaceous material

Process and apparatus for extracting biodiesel from algae.

PROCEDURE FOR THE PRODUCTION OF BIO ETHANOL AND FOR KOPRODUKTION OF ENERGY FROM A STÄRKEHALTIGEN VEGETABLE RAW MATERIAL

SESQUITERPENE SYNTHASEN AND BECOMING METHODS

PROCEDURE AND DEVICE FOR THE FERMENTIEREN OF ORGANIC MATERIAL

VERFAHREN ZUR BIOGENEN SYNTHESE VON METHAN AUS BIOMASSE

Es wird ein Verfahren zur biogenen Synthese von Methan aus Biomasse beschrieben, die durch eine Hydrolyse und eine Versäuerung aufgeschlossen und einer Methanogenese unterworfen.wird. Um eine Produktionssteigerung zu erreichen, wird vorgeschlagen, dass während der Versäuerung freigesetzte, überschüssige Wasserstoffionen in einem elektrischen Gleichfeld an einer Kathode gesammelt und als molekularer Wasserstoff abgezogen werden.

PROCEDURE FOR THE PRODUCTION OF METHANE FROM CELEBRATIONS PLANT MATERIALS.

PROCEDURE FOR THE PRODUCTION OF HETEROGENEOUS NACHBARDIHALOGENIERTEN PRODUCTS, THEIR APPLICATION AND NEW PRODUCTS.

PROCEDURE AND DEVICE FOR THE TREATMENT OF MUCK.

AQUEOUS POLYHYDROXYALKANOATDISPERSIONEN

Microbial synthesis of isoprenoid precursors, isoprenoids and derivatives including prenylated aromatics compounds

This disclosure generally relates to the use of enzyme combinations or recombinant microbes comprising same to make isoprenoid precursors, isoprenoids and derivatives thereof including prenylated aromatic compounds. Novel metabolic pathways exploiting Claisen, aldol, and acyioin condensations are used instead of the natural mevalonate (MVA) pathway or 1-deoxy-d-xylulose 5-phosphate (DXP) pathways for generating isoprenoid precursors such as isopentenyl pyrophosphate (IPP), dimethylallyl pyrophosphate (DMAPP), and geranyl pyrophosphate (GPP). These pathways have the potential for better carbon and or energy efficiency than native pathways. Both decarboxylative and non-carboxylative condensations are utilized, enabling product synthesis from a number of different starting compounds. These condensation reactions serve as a platform for the synthesis of isoprenoid precursors when utilized in combination with a variety of metabolic pathways and enzymes for carbon rearrangement and the addition ...

ABC transporters for the high efficiency production of rebaudiosides

Provided herein are genetically modified host cells, compositions, and methods for improved production of steviol glycosides. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleic acid expression cassette that expresses an ABC-transporter capable of transporting steviol glycosides to the extracellular space or to the luminal space of an intracellular organelle. In some embodiments, the host cell further comprises one or more heterologous nucleotide sequence encoding further enzymes of a pathway capable of producing one or more steviol glycosides in the host cell. The host cells, compositions, and methods described herein provide an efficient route for the heterologous production of steviol glycosides, including but not limited to, rebaudioside D and rebaudioside M.

A carotenoid preparation

Methods of producing influenza vaccine compositions

Methods and compositions for the optimization and production of influenza viruses, influenza B strains, in eggs and host cells suitable as influenza vaccines are provided.

Extraction of extracellular terpenoids from microalgae colonies

The invention provides methods of extracting and quantifying extracellular terpenoid hydrocarbons, e.g., botryococcenes, methylated squalenes, and carotenoids, from terpenoid-producing and secreting green microalgae.

PLANT ENZYMES FOR BIOCONVERSION

Fatty acid modification and tag assembly genes

The present invention relates to cells and methods for producing hydroxylated fatty acids. Also provided are novel polypeptides, and polynucleotides thereof, which can be used to produce the hydroxylated fatty acids, particularly in transgenic plants.

Cellulosic and lignocellulosic structural materials and methods and systems for manufacturing such materials by irradiation

CELLULOSIC AND LIGNOCELLULOSIC STRUCTURAL MATERIALS AND METHODS AND SYSTEMS FOR MANUFACTURING SUCH MATERIALS BY IRRADIATION Abstract Biomass (e.g. plant biomass, animal biomass, and municipal waste biomass) is 10 processed to produce useful products, such as fuels. For example, systems can use feedstock materials, such as cellulosic and/or lignocellulosic materials and/or starchy or sugary materials, to produce ethanol and/or butanol, e.g. by fermentation.

Hydrocarbon synthase gene, and use thereor

Provided is a hydrocarbon synthase gene coding a protein that has a new function with a superior ability to synthesize hydrocarbons such as alkane. The hydrocarbon synthase gene codes a protein which includes an amino acid sequence having the sequence motif represented by SEQ ID NO: 1, and which synthesizes, from an aldehyde compound, a hydrocarbon having one less carbon atom.

Production of isoprene, isoprenoid precursors, and isoprenoids using acetoacetyl-CoA synthase

This invention relates to a recombinant microorganism capable of producing isoprene and isoprene production with the use of such recombinant microorganism with good efficiency. In this invention, the acetoacetyl-CoA synthase gene encoding an enzyme capable of synthesizing acetoacetyl-CoA from malonyl-CoA and acetyl-CoA and one or more genes involved in isoprene biosynthesis that enables synthesis of isoprene from acetoacetyl-CoA are introduced into a host microorganism.

Production of acetyl-coenzyme a derived isoprenoids

Provided herein are compositions and methods for the heterologous production of acetyl-CoA-derived isoprenoids in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an acetaldehyde dehydrogenase, acetylating (ADA, E.C. 1.2.1.10) and an MEV pathway comprising an NADH-using HMG-CoA reductase. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an ADA and an MEV pathway comprising an acetoacetyl-CoA synthase. In some embodiments, the genetically modified host cell further comprises one or more heterologous nucleotide sequences encoding a phosphoketolase and a phosphotransacetylase. In some embodiments, the genetically modified host cell further comprises a functional disruption of the native PDH-bypass. The compositions and methods described herein provide an energy-efficient yet redox balanced route for the heterologous production of acetyl-CoA-derived ...

Fungi and products thereof

The present invention provides substantially purified or isolated fungi of Nodulisporium spp. (Dandenong Ranges isolate 1) or Ascocoryne spp. (Yarra Ranges isolates 7, 10, 1 1, 12, 13 and 15 and Otway Ranges isolates 1, 3, 4 and 5), plants infected with said fungi, organic compounds produced by said fungi, and related nucleic acids, polypeptides and methods.

Method of modulating metabolite biosynthesis in recombinant cells

Microbiological method for enriching petrol and petroleum products by isoparaffin and aromatic hydrocarbons with simultaneous removal of injurious additives andmeans for carrying out said method

FLAVOR AND FRAGRANCE ENZYMATIC ACTIVITIES OF ASPERGILLUS NIGER B1

METHOD OF MODULATING METABOLITE BIOSYNTHESIS IN RECOMBINANT CELLS

... ▓▓▓A method for modulating in a cell the expression of one or more genes involved ▓in the biosynthesis of a metabolite or a precursor for this metabolite by ▓inserting into a cell a nucleotide sequence coding for a transcription factor ▓comprising an AP2 DNA-binding domain, and/or by modifying the expression of a ▓nucleotide sequence coding for such a transcription factor already present in ▓the cell is provided. The method is useful for enhancing the biosynthesis of ▓secondary metabolites in plants such as alkaloids including terpenoid indole ▓alkaloids. Also provided is a method for enhancing stress tolerance in plants ▓by the use of such transcription factors.▓ ...

PHOSPHOLIPASES, NUCLEIC ACIDS ENCODING THEM AND METHODS FOR MAKING AND USING THEM

... ²²²The invention provides novel polypeptides having phospholipase activity, ²including, e.g., phospholipase A, B, C and D activity, patatin activity, ²phosphatidic acid phosphatases (PAP)) and/or lipid acyl hydrolase (LAH) ²activity, nucleic acids encoding them and antibodies that bind to them. ²Industrial methods, e.g., oil degumming, and products comprising use of these ²phospholipases are also provided.² ...

PROCESS FOR PRODUCING FATTY ACID AND ACTIVE INGREDIENT EXTRACTS, AND FATTY ACIDS AND ACTIVE INGREDIENTS THEMSELVES

The invention relates to a process for producing especially oleic acid and fatty acid extracts, and oleic acids and fatty acids themselves, according to the preamble of claims 1 and 11. In order to obtain active ingredients, especially oleic acids or fatty acids, from a noninvasive plant in large amounts here, bearing in mind, for reasons relating to agricultural laws, that this is achieved without use of genetic manipulation processes, it is proposed in accordance with the invention that the oleic acid or fatty acid extracts be obtained from the biological material, i.e. the plant material, of the novel plant variety CANDY (CPVO 2007/1958).

COMPOSITIONS AND METHODS FOR USING GENETICALLY MODIFIED ENZYMES

The disclosure relates to the biosynthesis of cannabinoids and related prenylated phenolic compounds using recombinant enzymes. In particular, the disclosure provides recombinant prenyltransferase enzymes engineered to produce a greater amount of a desired product, or to have a greater ability to catalyze a reaction using a desired substrate, as compared to the wild type prenyltransferase. The disclosure also provides methods of preparing such recombinant enzymes; as well as methods of use thereof in improving the biosynthesis of cannabinoids and related prenylated phenolic compounds.

GENES INVOLVED IN ISOPRENOID COMPOUND PRODUCTION

Genes have been isolated from Methylomonas 16a sp. encoding the isoprenoid biosynthetic pathway. The genes and gene products are the first isolated from a Methylomonas strain that is capable of utilizing single carbon (C1) substrates as energy sources. The genes and gene products of the present invention may be used in a variety of ways for the production of isoprenoid compounds in a variety of organisms.

CORYNEBACTERIUM GLUTAMICUM GENES ENCODING PROTEINS INVOLVED IN CARBON METABOLISM AND ENERGY PRODUCTION

Isolated nucleic acid molecules, designated SMP nucleic acid molecules, which encode novel SMP proteins from Corynebacterium glutamicum are described. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing SMP nucleic acid molecules, and host cells into which the expression vectors have been introduced. The invention still further provides isolated SMP proteins, mutated SMP proteins, fusion proteins, antigenic peptides and methods for the improvement of production of a desired compound from C. glutamicum based on genetic engineering of SMP genes in this organism.

METHOD FOR PRODUCING MONOTERPENOID COMPOUNDS

The invention relates to enzymes and methods for producing a monoterpenoid compound. In one aspect, the invention is a method for producing a monoterpenoid compound, comprising the steps of (1) providing a monoterpenoid precursor; (2) providing a NEPS enzyme; and (3) contacting the monoterpenoid precursor with the enzyme under catalytic conditions to produce an monoterpenoid compound.

MULTI-ZONE PROCESS AND APPARATUS FOR TREATING WASTEWATER

Wastewater containing organic matter may be treated using a multi-zone apparatus. In a first zone, organic matter in the wastewater may, among other things, be converted to at least volatile fatty acids (VFAs) and, thereafter, a portion of the treated wastewater may flow to a second zone that may, among other things, convert the VFAs to methane.

BIOLOGICAL DEGRADATION OF LOW-RANK COALS

Methods and processes for producing aerobic digestion products, such as organic acids, from a low-rank coal substrate are provided. Also provided are multistage bioreactor systems for carrying out the described methods and processes. In another aspect, product compositions comprising organic acids produced by the described methods and processes are provided, as well as methods for their use, including for the improvement of soil quality and/or plant growth.

IMPROVED PRODUCTION OF TERPENOIDS USING ENZYMES ANCHORED TO LIPID DROPLET SURFACE PROTEINS

Methods and expression systems are described herein that are useful for production of terpenes and terpenoids.

PRODUCTION OF ISOPRENOIDS

GENERATION OF MATERIALS WITH ENHANCED HYDROGEN CONTENT FROM ANAEROBIC MICROBIAL CONSORTIA

A microbial consortia for biogenically increasing the hydrogen content of a carbonaceous source material, where the consortia includes a first microbial consortium to metabolize the carbonaceous source material into one or more first intermediate hydrocarbons, a second microbial consortium, which includes one or more species of Pseudomonas microorganisms, to convert the first intermediate hydrocarbons into one or more second intermediate hydrocarbons and oxidized carbon, and a third microbial consortium to convert the second intermediate hydrocarbons into one or more smaller hydrocarbons and water, where the smaller hydrocarbons have a greater mol.% hydrogen than the carbonaceous source material.

POLYNUCLEOTIDES ENCODING ISOPRENOID MODIFYING ENZYMES AND METHODS OF USE THEREOF

... ²²²The present invention provides isolated nucleic acids comprising nucleotide ²sequences encoding isoprenoid modifying enzymes, as well as recombinant ²vectors comprising the nucleic acids. The present invention further provides ²genetically modified host cells comprising a subject nucleic acid or ²recombinant vector. The present invention further provides a transgenic plant ²comprising a subject nucleic acid. The present invention further provides ²methods of producing an isoprenoid compound, the method generally involving ²culturing a subject genetically modified host cell under conditions that ²permit synthesis of an isoprenoid compound modifying enzyme encoded by a ²subject nucleic acid.² ...

METHODS OF PROCESSING BIOMASS COMPRISING ELECTRON-BEAM RADIATION

The invention provides a method comprising converting a treated biomass feedstock to a product selected from the group consisting of alcohols, organic acids, sugars, hydrocarbons, and mixtures thereof, utilizing an enzyme and/or a microorganism, wherein the treated biomass feedstock has been prepared by irradiating a biomass feedstock with an electron beam at a dose rate of at least 1.0 Mrad/s for a time sufficient to deliver from about 10 Mrad to about 150 Mrad of radiation to the biomass feedstock so as to render the irradiated biomass feedstock more susceptible to chemical, enzymatic and/or biological attack than the biomass feedstock prior to electron beam treatment, the biomass feedstock having been physically treated prior to electron beam irradiation by grinding, milling or shearing. The invention may be used to process biomass into a product selected from the group consisting of alcohols, organic acids, sugars, hydrocarbons and mixtures thereof.

GAS DISTRIBUTOR FOR A STACKED PARTICLE BIOREACTOR

The invention relates to gas distributor (1) for a stacked particle biore actor which produces synfuels from biodegradable carbonaceous material compr ising a pipe having a series of apertures (3) through the wall of the pipe, with the series of apertures configured operatively to face downwards in the stacked particle bioreactor, and to be located in an indentation (2) in the outer wall of the pipe. The invention also extends to a stacked particle bi oreactor, method of forming a stacked particle bioreactor, and a method of p roducing gaseous fuel from biodegradable carbonaceous material using a stack ed particle which utilizes such a gas distributor.

MICROORGANISMS FOR BIOSYNTHESIS OF LIMONENE ON GASEOUS SUBSTRATES

Engineered microorganisms are provided that convert gaseous substrates, such as producer gas, into limonene. In some embodiments, limonene is pumped out of the cell via an efflux pump. In some embodiments, limonene, produced as described herein, is converted through catalytic dimerization into jet fuel. Producer gas used in the processes described herein for production of limonene may be derived from sources that include gasification of waste feedstock and/or biomass residue, waste gas from industrial processes, or natural gas, biogas, or landfill gas.

METHODS OF PROCESSING BIOMASS COMPRISING ELECTRON-BEAM RADIATION

This invention relates to a method of changing a molecular structure of a biomass feedstock. The method comprises processing a pretreated biomass feedstock to produce a product in which the processing comprises contacting with a microorganism and/or enzyme. The products are alcohols, organic acids, proteins, hydrocarbons, or sugars, and the pretreated biomass feedstock is obtained by pretreating a biomass feedstock using two or more different pretreatment methods that each change the molecular structure of the biomass feedstock. The different pretreatment method is radiation, sonication, pyrolysis, and/or oxidation.

RECOMBINANT CELL, AND METHOD FOR PRODUCING ISOPRENE

The purpose of the present invention is to provide a series of technologies capable of producing isoprene from a synthesis gas, etc. Provided is a recombinant cell capable of producing isoprene from at least one C1 compound, wherein: a nucleic acid that codes an isoprene synthase has been introduced to a host cell having the ability to synthesize isopentenyl diphosphate by using a non-mevalonate pathway; the nucleic acid is expressed inside the host cell; and said C1 compound is selected from a group comprising carbon monoxide, carbon dioxide, formic acid, and methanol. The host cell is exemplified by a Clostridium cell or a Moorella cell. Also provided is a production method for isoprene using this recombinant cell.

METHODS FOR STABILIZING PRODUCTION OF ACETYL-COENZYME A DERIVED COMPOUNDS

The present disclosure relates to the use of a switch for the production of heterologous non-catabolic compounds in microbial host cells. In one aspect, provided herein are genetically modified microorganisms that produce non-catabolic compounds more stably when serially cultured under aerobic conditions followed by microaerobic conditions, and methods of producing non-catabolic compounds by culturing the genetically modified microbes under such culture conditions. In another aspect, provided herein are genetically modified microorganisms that produce non-catabolic compounds more stably when serially cultured in the presence of maltose followed by the reduction or absence of maltose, and methods of producing non-catabolic compounds by culturing the genetically modified microbes under such culture conditions.

INTEGRATED BIO-DIGESTION FACILITY

Methods and systems for pro-cessing organic waste material are provided. These methods and systems include integrating an anaerobic bio-digester and nutrient recovery module with a bio-production facility, which can locally provide feedstock for the bio-pro-duction facility, and can locally provide organ-ic material for the anaerobic bio-digester. Methods and systems for integrating an anaer-obic bio- digester with a gas cleaner are also provided, which can recover nutrients while cleaning the biogas produced by the anaerobic bio-digester.

LIGNOCELLULOSIC BIOMASS CONVERSION

The present invention relates to a process for the production of second generation biofuels and/or sugar based chemicals - for example ethanol, butanol etc - and/or materials - for example plastics, single cell proteins etc. - together with sulfonated lignin from lignocellulosic biomass, in particular from lignocellulosic biomass comprising, among others, annual plants, agricultural waste, or wood. In particular, the present invention relates to a process for the production of sugar based chemicals, biofuels or materials together with sulfonated lignin from lignocellulosic biomass comprising the pretreatment of a lignocellulosic biomass in a sulfite cooking step.

NOVEL STRAINS OF MICROALGAE OF THE BOTRYOCOCCUS GENUS, AND METHOD FOR CULTIVATING SAID MICROALGAE IN A MIXOTROPHIC MODE

L'invention se rapporte à de nouvelles souches de microalgues appartenant au genre Botryococcus, capables de croître en mode mixotrophe, ainsi qu'à un procédé de culture faisant appel à un apport de lumière sous forme de flashs pour la production de lipides et d'hydrocarbures, notamment sous forme de botryococcènes, utiles pour la production de biocarburants.

PROCESS FOR GENERATING A HYDROCARBON FEEDSTOCK FROM LIGNIN

The present invention discloses processes for generating a hydrocarbon feedstock for biofuels synthesis from lignin via hydroprocessing. Embodiments of the present invention can occur in a refinery setting or in a paper mill setting. Embodiments of the present invention can utilize the separated lignin or the entire black liquor solution.

METHOD FOR THE PREPARATION AND EXTRACTION OF SQUALENE FROM MICROALGAE

L'invention se rapporte à un de production de squalène par des microalgues appartenant à la famille des Thraustochytriales sp., de préférence à des teneurs entre 2 et 12 g pour 100 g de biomasse sèche, caractérisé en ce qu'il comprend les étapes consistant à cultiver des microalgues appartenant à la famille des Thraustochytriales sp. à une température comprise entre 25 et 35°C, de préférence comprise entre 28 et 32°C, plus préférentiellement de l'ordre de 30°C et ajouter dans le milieu de culture 1 à 1000 µg de vitamine B12 par litre de milieu de culture.

METHOD FOR EXTRACTING SQUALENE FROM MICROALGAE

L'invention se rapporte à un procédé d'extraction sans solvant organique de squalène produit par fermentation de microalgues appartenant à la famille des Thraustochytriales sp., caractérisé en ce qu'il comprend les étapes suivantes : 1) préparer une biomasse de microalgues appartenant à la famille des Thraustochytriales de manière à réduire la concentration en solubles interstitiels et atteindre ainsi une pureté comprise entre 30 et 99 % exprimée en poids sec de biomasse sur poids sec total du milieu de fermentation, 2) traiter la biomasse ainsi obtenue à l'aide d'une enzyme de type- protéases choisies dans le groupe des protéases netres ou basiques de manière à rompre la paroi cellulaire desdites microalgues tout en prévenant la formation de l'émulsion produite par ledit traitement enzymatique, 3) centrifuger le mélange réactionnel ainsi obtenu afin de séparer l'huile de la phase aqueuse, et 4) récupérer l'huile brute enrichie en squalène ainsi produite.

DITERPENE PRODUCTION

The present invention relates to a recombinant microorganism comprising one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity; a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity, whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol. The recombinant microorganism may also be capable of expressing one or more UDP-glucosyltransferases such that the microorganism is capable of producing one or more steviol glycosides.

SYSTEM AND PROCESS FOR BIOMASS CONVERSION TO RENEWABLE FUELS WITH BYPRODUCTS RECYCLED TO GASIFIER

This invention relates generally to a method and system for improving the conversion of carbon-containing feed stocks to renewable fuels, and more particularly to a thermal chemical conversion of biomass to renewable fuels and other useful chemical compounds, including gasoline and diesel, via a unique combination of unique processes. More particularly, this combination of processes includes (a) a selective pyrolysis of biomass, which produces volatile hydrocarbons and a biochar; (b) the volatile hydrocarbons are upgraded in a novel catalytic process to renewable fuels, (c) the biochar is gasified at low pressure with recycled residual gases from the catalytic process to produce synthesis gas, (d) the synthesis gas is converted to dimethyl ether in a novel catalytic process, and (e) the dimethyl ether is recycled to the selective pyrolysis process.

ENZYME PRODUCTION COMPOSITIONS AND METHODS

The present invention provides compositions and methods for the production of enzymes. Interest has arisen in fermentation of carbohydrate-rich biomass to provide alternatives to petrochemical sources for fuels and organic chemical precursors. "First generation" bioethanol production from carbohydrate sources (e.g., sugar cane, com, wheat, etc.) have proven to be marginally economically viable on a production scale.

PROCEDURE FOR THE PRETREATMENT OF ORGANIC WASTES FROM SLAUGHTERINGS.

METHOD FOR SELECTIVE PRODUCTION OF HYDROCARBON FUELS FROM ALGAE USING WATER AT UNDER SUBCRITICAL CONDITIONS

TREATMENT OF BIOMASS

СПОСОБ ФЕРМЕНТАЦИИ НИЗКОМОЛЕКУЛЯРНОГО САХАРА В ПРОДУКТ

Биомассу (например, растительную биомассу, животную биомассу и биомассу муниципальных отходов) обрабатывают для использования в производстве полезной продукции, включая топливо. Например, системы могут использовать материал биомассы, в том числе целлюлозные и/или лигноцеллюлозные материалы, для повышения производства продукции, например производства этанола и/или бутанола, путем ферментации.

БИОДИЗЕЛЬНОЕ ТОПЛИВО

Способ получения биодизельного топлива, включающий: (I) получение бионефти, содержащей по меньшей мере один глицерид жирной кислоты; (II) контактирование бионефти по меньшей мере с одним микроорганизмом, способным гидролизовать по меньшей мере одну эфирную связь указанного по меньшей мере одного глицерида для получения гидролизованной бионефти; (III) прямое или косвенное использование указанной гидролизованной бионефти в биодизельном топливе.

PRODUCTION OF DITERPENA

PRODUCTION OF DITERPENA

MICROORGANISMS AND ARTIFICIAL ECOSYSTEM FOR PRODUCTION OF PROTEIN, FOOD PRODUCTS AND MINERAL BY-PRODUCTS SUBSTRATES C1

СПОСОБ И БИОРЕАКТОР ДЛЯ ПОЛУЧЕНИЯ СИНТЕТИЧЕСКОГО ТОПЛИВА ИЗ УГЛЕРОДСОДЕРЖАЩЕГО МАТЕРИАЛА

Предлагается способ продуцирования топлива из биодеградируемого углеродсодержащего материала с использованием многослойного биореактора из частиц. Многослойный биореактор из частиц сформирован из частиц, включающих биодеградируемый углеродсодержащий материал. Биодеградируемый углеродсодержащий материал в многослойном биореакторе из частиц аэробно и/или анаэробно биоконвертируется в одно или несколько синтетических топлив, которые собирают из реактора. Синтетические топлива, продуцированные с помощью способа, могут включать синтетическую нефть, спирт и/или газообразное топливо, содержащее метан. Предпочтительно способ включает фазу аэробной биообработки с последующей фазой анаэробной биоконверсии. Также описан многослойный биореактор из частиц для проведения анаэробной и предпочтительно аэробной деградации.

БИОСИНТЕЗ 1-АЛКЕНОВ В СКОНСТРУИРОВАННЫХ МИКРООРГАНИЗМАХ

Различные 1-алкены, в том числе 1-нонадецен и 1-октадецен, синтезированы посредством сконструированных микроорганизмов и способов по данному изобретению. В некоторых вариантах осуществления данные микроорганизмы содержат рекомбинантные 1-алкенсинтазы. Сконструированные микроорганизмы могут представлять собой фотосинтезирующие микроорганизмы, такие как цианобактерии.

СПОСОБЫ И КОМПОЗИЦИИ ДЛЯ РЕКОМБИНАНТНОГО БИОСИНТЕЗА H-АЛКАНОВ

В настоящем описании раскрыты способы и композиции для модификации фотоавтотрофных организмов в качестве организмов-хозяев, которые в результате эффективно превращают углекислый газ на свету в н-алканы, и, в частности, применение таких организмов для коммерческого производства н-алканов и подобных молекул.

СПОСОБ И АППАРАТ ДЛЯ ПРОИЗВОДСТВА БИОЭТАНОЛА И ДРУГИХ ПРОДУКТОВ ФЕРМЕНТАЦИИ

Способ преобразования органического материала в углеводородное топливо, такое как этанол, способ включает процесс ферментации, при котором ферментируют органический материал, таким образом обеспечивая ферментативный бульон, процесс разделения, при котором разделяют ферментированный материал на углеводородное топливо и остаточный продукт, процесс преобразования, при котором, по меньшей мере, частично преобразовывают остаточный продукт в энергию, и процесс распределения энергии, при котором распределяют по меньшей мере часть энергии, полученной в процессе преобразования, на процесс ферментации.

ВЫРАЩИВАНИЕ МОРСКИХ ВОДОРОСЛЕЙ ДЛЯ БИОТОПЛИВА

Способ получения водорослевого масла, включающий в себя этапы регулирования роста для обеспечения интенсивного роста для получения начальных средств для выращивания водорослей. Этап выращивания водорослей с использованием преимущественно солнечного света. Этап обработки водорослей, полученных из выращенных водорослей, предпочтительно при помощи процесса влажного извлечения; и причем по меньше мере один из этапов включает в себя применение мешка, выполненного с возможностью присоединения к нему потоков газа или жидкости, состоящих по меньшей мере из одного из воды, CO2, кислорода или воздуха.

СПОСОБ ПОЛУЧЕНИЯ БИОГАЗА ПРИ ИЗГОТОВЛЕНИИ ЦЕННОГО ПРОДУКТА, В ЧАСТНОСТИ, КРАХМАЛА ИЗ МУКИ ИЗ ЗЕРНА ЗЛАКОВЫХ КУЛЬТУР

Изобретение относится к способу получения биогаза при изготовлении ценного продукта, в частности, крахмала и/или белка из муки из зерна злаковых культур, при котором муку смешивают со свежей или технологической водой в тесто, тесто разделяют как минимум на две фракции, в частности, под действием центробежных сил на тяжелую фракцию первичного крахмала, фракцию белка и вторичного крахмала (фазу, которая выпускается через сопла декантатора), и фракцию пентозана. Как минимум из одной фракции, полученной при разделении, получают биогаз, используемый для получения энергии, причем фракцию, используемую для получения биогаза, подвергают как минимум одной операции сжижения (505) и разделения фаз (506), причем биогаз получают из жидкой фазы разделенных фаз (506).

УСТАНОВКА ДЛЯ ПОЛУЧЕНИЯ БИОГАЗА

Установка для получения биогаза содержит корпус-сборник, исполненный без днища, донный шнек, понтоны, автономный источник энергии, редуктор, привод донного шнека и устройство для регулирования глубины взбалтывания ила. Взбалтывание речного ила осуществляют донным шнеком. Улавливание биогаза из толщи воды осуществляют сборником без днища.

УСТАНОВКА ДЛЯ ОПРЕДЕЛЕНИЯ МЕТАНОГЕННЫХ ПОТЕНЦИАЛОВ БИОМАСС В ЛАБОРАТОРНЫХ УСЛОВИЯХ

Установка для определения метаногенных потенциалов биомасс в лабораторных условиях выполнена в виде основы, термоизолированного с подогревом инкубатора, ферментеров, ванны с водой, герметизированной емкости, герметических полиэтиленовых пакетов, горизонтальной трубки, вертикального цилиндра, штока, электромагнитного механизма поднимания штока, блока управления и записи. На основе установлен термоизолированный с подогревом инкубатор, а в инкубаторе размещены ферментеры, изготовленные из газонепроницаемого полиэтилена и заправленные биомассами и затравками. Из ферментеров уделен воздух и установлено газоотводные трубки с зажимами на конце, а ферментеры пронумерованы. На основе установлена ванна с водой и герметизированная емкость, а в емкости установлены герметические полиэтиленовые пакеты с входными патрубками, выведенными наружу емкости.

PRODUCTION OF SQUALENE USING YEAST

СПОСІБ ОТРИМАННЯ БІОГАЗУ ІЗ СИНЬОЗЕЛЕНИХ ВОДОРОСТЕЙ

Спосіб отримання біогазу із синьозелених водоростей включає збір та використання концентрованої біомаси ціанобактерій, зібраних під час "цвітіння" з акваторії водосховищ дніпровського каскаду, для отримання клар-газу. Отримують клар-газ за біотехнологією метанового "бродіння".

Способ деструктивной переработки горючего сланца

Использование: в химической промышленности для получения продукта, используемого в качестве топлива. Задача: повышение выхода летучих продуктов (газов) и смолы с единицы массы исходного сырья. Сущность изобретения : материал сланца дробят до класса крупности 100% -2 мм, заливают водой и обрабатывают автотрофными и/или гетеротрофными бактериями, затем обезвоживают и сушат при температуре 105°C. Полученный кек подвергают двухстадийному пиролизу: на первой стадии до температуры 300°C, отгоняют смолу и часть газов; на второй стадии, при дальнейшем нагреве до 650°C, отгоняют оставшуюся часть газов. 1 н. п. ф-лы, 2 з. п. ф-лы, 6 прим., 4 табл.

ФЕРМЕНТАТОР БІОГАЗУ

Винахід належить до поновлюваних джерел енергії і може бути використаний у сільському господарстві. Ферментер біогазу має ізольований робочий об'єм метанового бродіння, насоси завантаження, придонного перемішування біомаси і вивантаження старого мулу, засоби плавучості і термостатування. Герметизований газозбірник виконано світлопрозорим для видимого спектра і обладнано обертовим іммобілізатором метанових мікроорганізмів з щілинними світло- і газопідводами, розміщеними на межі газ-суспензія, а насос завантаження обладнано бункером і шнековими подавачами мулу чи біомаси.

СПОСОБ ПОЛУЧЕНИЯ ИММОБИЛИЗОВАННЫХ МИКРООРГАНИЗМОВ

Изобретение относится к способу получения иммобилизованных микроорганизмов, включающему облучение волокнистого материала пучком электронов в атмосфере воздуха при полной дозе, составляющей по меньшей мере 5 Мрад, для получения функционализированного волокнистого материала, имеющего карбоксильные группы, и обеспечение контактирования указанного функционализированного волокнистого материала с микроорганизмами, например дрожжами и/или бактериями.

CELLULOSE AND LIGNOCELLULOSE MATERIALS AND METHODS AND SYSTEMS FOR PRODUCTION OF SUCH MATERIALS IRRADIATION

METHOD FOR BIOGAS PRODUCTION OF BLUE-GREEN ALGAE

Nucleic acid molecule

The invention relates to an isolated nucleic acid molecule encoding a polypeptide capable of producing a triterpenoid hydrocarbon. The invention also relates to the encoded polypeptide, a vector comprising the nucleic acid molecule, a recombinant non-human organism comprising the nucleic acid molecule, and to methods of producing a triterpenoid hydrocarbon or an intermediate of biofuel using the nucleic acid molecule, polypeptide or recombinant organism.

Filamentous fungi and methods for producing trichodiene from lignocellulosic feedstocks

The present invention relates to the production of a C-15 fuel from lignocellulosic or other feedstock. Specifically at least double mutant of filamentous fungi having the isoprenoid pathway results in production of trichodiene in commercial quantities. One embodiment of the invention relates to producing the fuel at the site of the lignocellulosic feedstock to reduce costs of shipping the feedstock.

Novel Sesquiterpene Synthase Gene and Protein

The invention relates to sesquiterpene synthases and methods for their production and use. Particularly, the invention provides nucleic acids comprising the nucleotide sequence of citrus valencene synthase (CVS) which codes for at least one CVS. The invention further provides nucleic acids comprising the nucleotide sequence coding for amino acid residues forming the tier 1 and tier 2 domains of CVS. The invention also provides for methods of making and using the nucleic acids and amino acids of the current invention.

Chimeric isoprenoid synthases and uses thereof

Provided is a chimeric isoprenoid synthase polypeptide including a first domain from a first isoprenoid synthase joined to a second domain from a second, heterologous, isoprenoid synthase, whereby the chimeric isoprenoid synthase is capable of catalyzing the production of isoprenoid reaction products that are not produced in the absence of the second domain of the second, heterologous, isoprenoid synthase. Also provided is a chimeric isoprenoid synthase polypeptide including an asymmetrically positioned heterologous domain, whereby the chimeric isoprenoid synthase is capable of catalyzing the production of isoprenoid reaction products that are not produced when the domain is positioned at its naturally-occurring site in the isoprenoid synthase polypeptide. 1. A method for producing a chimeric isoprenoid synthase polypeptide , comprising:(a) providing a cell comprising DNA encoding the chimeric isoprenoid synthase polypeptide, wherein the encoded synthase comprises a first sesquiterpene isoprenoid synthase polypeptide domain joined to a second, different, sesquiterpene isoprenoid synthase polypeptide domain such that the chimeric isoprenoid synthase polypeptide produces one or more isoprenoid products that are not produced in the absence of the second domain of the second synthase;(b) culturing said cell under conditions for expressing the DNA to produce the chimeric isoprenoid synthase; and(c) recovering the chimeric isoprenoid synthase.2. The method of claim 1 , wherein the encoded chimeric isoprenoid synthase comprises a first isoprenoid sesquiterpene synthase polypeptide domain joined to a second different isoprenoid sesquiterpene synthase polypeptide domain claim 1 , interrupted by or including a ratio determinant domain claim 1 , whereby the chimeric isoprenoid sesquiterpene synthase polypeptide encoded by the DNA catalyzes the production of at least one isoprenoid sesquiterpene synthase reaction product that is not produced in the absence of the second ...

COMPOSITIONS AND METHODS FOR PRODUCING ISOPRENE

The invention features methods for producing isoprene from cultured cells. The invention also provides compositions that include these cultured cells. 16-. (canceled)7. A method of producing isoprene , the method comprising (i) an isoprene synthase polypeptide,', '(ii) an isopentenyl-diphosphate delta-isomerase (IDI) polypeptide, and', '(iii) at least one of a 1-Deoxyxylulose-5-phosphate synthase (DXS) polypeptide and/or one or more mevalonate (MVA) pathway polypeptides and', 'wherein the cells', {'sub': 'wcm', '(1) produce greater than about 400 nmole/g/hr of isoprene,'}, '(2) convert more than about 0.002 molar percent of the carbon that the cells consume from a cell culture medium into isoprene,', {'sub': 'broth', '(3) have an average volumetric productivity of isoprene greater than about 0.1 mg/L/hr of isoprene, or'}, {'sub': 'dcm', '(4) produce at least 0.0037 grams of isoprene per gram of dry cell mass (g); and'}], '(a) culturing recombinant cells under suitable culture conditions for the production of isoprene, wherein the cells comprise one or more nucleic acids encoding'}(b) producing isoprene.8. The method of claim 7 , further comprising recovering the isoprene produced by the cells.910-. (canceled)11. The method of claim 7 , wherein the nucleic acid encoding an isoprene synthase polypeptide of (a)(i) is a heterologous nucleic acid encoding an isoprene synthase polypeptide.12. The method of claim 7 , wherein the nucleic acid encoding an isoprene synthase polypeptide of (a)(i) is a copy of an endogenous nucleic acid encoding an isoprene synthase polypeptide.13. The method of claim 7 , wherein the isoprene synthase polypeptide is a plant isoprene synthase polypeptide.14. The method of claim 13 , wherein the plant isoprene synthase polypeptide is a poplar isoprene synthase polypeptide.15. The method of claim 13 , wherein the plant isoprene synthase polypeptide is a kudzu isoprene synthase polypeptide.16. The method of claim 7 , wherein the nucleic acid ...

UTILIZATION OF PHOSPHOKETOLASE IN THE PRODUCTION OF MEVALONATE, ISOPRENOID PRECURSORS, AND ISOPRENE

The invention provides for methods for the production of mevalonate, isoprene, isoprenoid precursor molecules, and/or isoprenoids in cells via the heterologous expression of phosphoketolase enzymes. 1. Recombinant cells capable of producing of isoprene , wherein the cells comprise one or more heterologous nucleic acids encoding a polypeptide having phosphoketolase activity and (i) one or more nucleic acids encoding one or more polypeptides of the complete MVA pathway and (ii) a heterologous nucleic acid encoding an isoprene synthase polypeptide , wherein culturing of said recombinant cells in a suitable media provides for the production of isoprene.2. The cells of claim 1 , wherein the one or more heterologous nucleic acids encoding a polypeptide having phosphoketolase activity is capable of synthesizing glyceraldehyde 3-phosphate and acetyl phosphate from xylulose 5-phosphate.3. The cells of claim 1 , wherein the one or more heterologous nucleic acids encoding a polypeptide having phosphoketolase activity is capable of synthesizing erythrose 4-phosphate and acetyl phosphate from fructose 6-phosphate.4Bifidobacterium longum, Enterococcus galliniarum, Clostridium acetobutilicum, Nostoc punctiforme, Rhodopseudomonas palustris, Pantoea, Mucilaginibacter paludis, Thermobifida fusca, Bifidobacterium breve, Rahnella aquatili, Bifidobacterium animalis, Gardnerella vaginalis, Streptomyces avermitilis, Lactobacillus plantarumLactobacillus reuteri.. The cells of claim 1 , wherein the heterologous nucleic acid encoding a polypeptide having phosphoketolase activity is selected from the group consisting of: claim 1 , and5Bifidobacterium longum, EnterococcusClostridium acetobutilicum.. The cells of claim 1 , wherein the heterologous nucleic acid encoding a polypeptide having phosphoketolase activity is selected from the group consisting of: galliniarum claim 1 , and6. The cells of claim 1 , the heterologous nucleic acid encoding an isoprene synthase polypeptide is a plant ...

PRODUCTION OF MONOTERPENES

The present invention relates to methods for producing monoterpenes, particularly tricyclene, by culturing microbial organisms that express a terpene synthase and optionally a prenyl transferase. 1. A method for producing tricyclene comprising:culturing a microbial organism expressing a heterologous terpene synthase under conditions in which the terpene synthase converts geranyl diphosphate to tricyclene, wherein the tricyclene is produced at a level of at least 0.5% of total monoterpene production by the organism.2. The method according to claim 1 , wherein the terpene synthase is a bornyl diphosphate synthase or a variant thereof.3. The method according to claim 1 , wherein the terpene synthase has at least 80% sequence identity to SEQ ID NO: 2 or 4.4. (canceled)5. The method according to claim 2 , wherein the terpene synthase comprises at least 90% sequence identity to SEQ ID NO: 2 claim 2 , and comprises an amino acid substitution at one or both of positions V399 and I404 of SEQ ID NO: 2.6. The method according to claim 5 , wherein the substitution at position V399 is V399I.7. The method according to claim 5 , wherein the terpene synthase further comprises an amino acid substitution at one or more of positions S4 claim 5 , E159 claim 5 , G338 claim 5 , S267 claim 5 , I291 claim 5 , I297 claim 5 , K285 claim 5 , T460 claim 5 , and F525 of SEQ ID NO: 2.8. The method according to claim 1 , wherein the terpene synthase is a camphene synthase or a variant thereof.9. The method according to claim 8 , wherein the terpene synthase has at least 80% sequence identity to SEQ ID NO: 6 or 8.10. (canceled)11. The method according to claim 8 , wherein the terpene synthase comprises at least 90% sequence identity with SEQ ID NO: 6 claim 8 , and comprises an amino acid substitution at one or more of positions N18 claim 8 , A283 claim 8 , I320 claim 8 , and T431 of SEQ ID NO: 6.12. The method according to claim 11 , wherein the terpene synthase further comprises an amino acid ...

(r)-hydroxynitrile lyase from brassicaceae

The invention concerns a polypeptide which can be isolated from the Brassicaceae family and which has at least the activity of a hydroxynitrile lyase (HNL). The hydroxynitrile lyase of the invention is the first HNL from the Brassicaceae family. The plants ( Arabidopsis ) from which this enzyme or its gene is isolated is also described as non-cyanogenic. All HNL-containing plants described so far are cyanogenic plants and so it has until now been assumed that only cyanogenic plants contain hydroxynitrile lyases. Surprisingly, it transpires that a polypeptide (AtHNL) of the invention is (R)-selective. The amino acid sequence gives a theoretical molecular weight of 29.2 kDa for the AtHNL subunit. The calculated molecular mass of the protein of approximately 30 kDa can be confirmed by SDS gel electrophoresis.

Heat-Stable Persephonella Carbonic Anhydrases and Their Use

The present invention relates to use of Persephonella carbonic anhydrase in CO 2 extraction, e.g., from flue gas, natural gas, biogas or ambient air. The Persephonella carbonic anhydrases are especially well suited for these purpose due to their extreme thermostability.

NOVEL ALDOLASE AND PRODUCTION PROCESS OF SUBSTITUTED ALPHA-KETO ACIDS

4-(Indol-3-ylmethyl)-4-hydroxy-2-oxoglutarate, which is useful as an intermediate in the synthesis of monatin, may be synthesized from indole pyruvic acid and pyruvic acid (and/or oxaloacetic acid) by using a novel aldolase derived from the genus or 1. An isolated polynucleotide of following (a) or (b):(e) a polynucleotide having the nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15; or(f) a polynucleotide that hybridizes under stringent conditions with a polynucleotide having the nucleotide sequence complementary to the full-length nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15, and encodes a protein having aldolase activity, wherein said stringent conditions comprise a wash step in 0.1×SSC and 0.1% SDS at 65° C.2. An isolated polynucleotide of following (c) or (d):(g) a polynucleotide that encodes a protein comprising the amino acid sequence of SEQ ID NO: 16; or(h) a polynucleotide that encodes a protein having an amino acid sequence that contains a substitution, deletion, insertion, or addition of one to ten amino acid residues in the amino acid sequence of SEQ ID NO: 16, and has aldolase activity.3. A recombinant DNA produced by ligating into a vector DNA a polynucleotide selected from the group consisting of:(e) a polynucleotide having the nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15;(f) a polynucleotide that hybridizes under stringent conditions with a polynucleotide having the nucleotide sequence complementary to the full-length nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15, and encodes a protein having aldolase activity, wherein said stringent conditions comprise a wash step in 0.1×SSC and 0.1% SDS at 65° C.;(g) a polynucleotide that encodes a protein comprising the amino acid sequence of SEQ ID NO: 16; and(h) a ...

ENHANCED PRODUCTION OF ISOPRENE USING HOST CELLS HAVING DECREASED ISPA ACTIVITY

This invention relates to recombinant microorganisms capable of producing isoprene and isoprene production with the use of such recombinant microorganism with good efficiency. In this invention, functional activity of the ispA gene is altered to reduce the production of isoprenoid molecules in recombinant cells engineered to produce isoprene or in cells otherwise susceptible to isoprenoid accumulation during fermentation. This decreased ispA gene functional activity enables enhanced synthesis of isoprene in a host microorganism. 1. Recombinant cells capable of producing of isoprene , wherein said cells comprise an ispA gene having decreased functional activity and one or more nucleic acids encoding:(a) an isoprene synthase polypeptide, wherein the isoprene synthase polypeptide is encoded by a heterologous nucleic acid; and(b) one or more mevalonate (MVA) pathway polypeptides,wherein culturing of said recombinant cells in a suitable media provides for the production of said polypeptides and synthesis of isoprene.2. The recombinant cells of claim 1 , wherein the functional activity of the ispA gene is decreased by:a. deleting the ispA gene;b. decreasing ispA gene expression;c. decreasing ispA protein activity;d. decreasing ispA protein expression; ore. temporally modulating ispA expression.3. The recombinant cells of claim 2 , wherein ispA gene expression is decreased by placing the ispA gene under the control of a weak promoter.4. The recombinant cells of claim 2 , wherein ispA gene expression is decreased by placing the ispA gene under the control of an auto-regulatory promoter.5. The recombinant cells of claim 2 , wherein ispA protein activity is decreased by translational fusion of the ispA protein with a proteolytic tag.6. The recombinant cells of claim 2 , wherein ispA protein activity is decreased by use of antisense RNA.7. The recombinant cells of claim 2 , wherein ispA protein activity is decreased by introducing one or more mutations into a ribosomal binding ...

PRODUCTION OF ISOPRENE UNDER REDUCED OXYGEN INLET LEVELS

This invention relates to methods for producing isoprene by culturing recombinant cells (e.g., cells engineered to produce isoprene) under reduced oxygen inlet levels. 1. A method for producing isoprene comprising (a) culturing a recombinant host cell comprising a heterologous nucleic acid encoding isoprene synthase under reduced oxygen inlet levels wherein the cell is in fermentation or production phase; and (b) producing isoprene.2. The method of further comprising recovering the isoprene.3. The method of wherein the reduced oxygen inlet level is between about 5% to about 15% oxygen.4. The method of wherein the reduced oxygen inlet level is between about 7% to about 10% oxygen.5. The method of wherein the reduced oxygen inlet level is about 7.7% oxygen.6. The method of wherein the reduced oxygen inlet level is about 9.3% oxygen.7. The method of wherein the isoprene synthase is a plant isoprene synthase.8. The method of claim 7 , wherein the plant isoprene synthase is a poplar isoprene synthase claim 7 , a kudzu isoprene synthase claim 7 , a willow isoprene synthase claim 7 , or a eucalyptus isoprene synthase.9PuerariaPopulusPopulus alba×Populus tremula.. The method of wherein the plant isoprene synthase is an isoprene synthase from or or a hybrid claim 7 ,10Pueraria montanaPueraria lobata, Populus tremuloides, Populus alba, Populus nigraPopulus trichocarpa.. The method of wherein the plant isoprene synthase polypeptide is selected from the group consisting of or claim 7 , and11. The method of wherein the isoprene synthase is an isoprene synthase variant.12. The method of wherein the cell further comprises a heterologous nucleic acid encoding for one or more MVA pathway polypeptide and/or one or more DXP pathway polypeptide.13. The method of claim 12 , wherein the cell further comprises a heterologous nucleic acid encoding for one or more IDI polypeptide.14. The method of claim 12 , wherein any one or more copies of a heterologous nucleic acid is overexpressed.15. ...

POLYPEPTIDE HAVING DITERPENE SYNTHASE ACTIVITY

The present application, among others, relates to novel polypeptides having diterpene synthase activity, nucleic acid molecules encoding same, as well as to a gene cluster from which is thought to be involved in the biosynthetic pathway for producing pleuromutilin. 115.-. (canceled)16. An isolated polypeptide ,the polypeptide comprising an amino acid sequence which comprisesa sequence having at least 50% sequence identity to SEQ ID NO: 1,a sequence having at least 40% sequence identity to SEQ ID NO: 2, and i) a sequence having at least 15% sequence identity to SEQ ID NO: 7;', 'ii) a sequence having at least 25% sequence identity to SEQ ID NO: 4;', 'iii) a sequence having at least 45% sequence identity to SEQ ID NO: 5; and', 'iv) a sequence having at least 45% sequence identity to SEQ ID NO: 6,, 'at least one sequence selected from the group consisting of'}{'i': 'Clitopilus passeckerianus', 'wherein SEQ ID NOs: 1-2 and 4-7 are of origin and wherein said polypeptide is a diterpene synthase.'}17Clitopilus passeckerianus. The isolated polypeptide according to claim 16 , wherein said amino acid sequence further comprises a sequence having at least 50% sequence identity to SEQ ID NO: 3 claim 16 , wherein SEQ ID NO: 3 is of origin.18. The isolated polypeptide according to claim 16 , wherein the molecular weight of the polypeptide is between 90 kDa and 140 kDa.19. The isolated polypeptide according to claim 16 , wherein the polypeptide comprises an amino acid sequence which amino acid sequence comprises a sequence having at least 70% sequence identity to SEQ ID NO: 9.20. The isolated polypeptide according to claim 16 , wherein the polypeptide is capable of catalyzing the conversion of geranyl pyrophosphate into a pleuromutilin precursor.21. The isolated polypeptide according to claim 20 , wherein said pleuromutilin precursor is a compound according to formula (I).22. The isolated polypeptide according to claim 16 , wherein said polypeptide is derivable from a fungal host. ...

MODIFIED ETHYLENEDIAMINE-N, N'-DISUCCINATE: ETHYLENEDIAMINE LYASE

The present invention provides a modified ethylenediamine-N,N′-disuccinate:ethylenediamine lyase. The present invention also provides a protein that comprises the amino acid sequence represented by SEQ ID NO: 1; or a protein that comprises an amino acid sequence derived from the amino acid sequence represented by SEQ ID NO: 1 by deletion, substitution, or addition of one or more amino acid residues, and has an ethylenediamine-N,N′-disuccinate:ethylenediamine lyase activity. 153-. (canceled)54. An isolated gene encoding a protein comprising the amino acid sequence of SEQ ID NO:1 and having an ethylenediamine-N ,N′-disuccinate:ethylenediamine lyase activity.55. An isolated gene encoding a protein comprising the amino acid sequence of SEQ ID NO:1 having 1 to 10 substitutions , and wherein the protein has ethylenediamine-N ,N′-disuccinate:ethylenediamine lyase activity that is heat resistant.56. The isolated gene of claim 55 , wherein one substitution is the substitution of the lysine residue at position 120 with a glutamic acid residue.56. The isolated gene of claim 55 , wherein one substitution is the substitution of the isoleucine residue at position 166 with a serine residue.58. The isolated gene of claim 55 , wherein one substitution is the substitution of the isoleucine residue at position 166 with a threonine residue.59. The isolated gene of claim 55 , wherein one substitution is the substitution of the alanine residue at position 365 with a valine residue.60. An isolated gene encoding a protein comprising the amino acid sequence of SEQ ID NO:1 having 2 to 10 substitutions claim 55 , wherein the substitutions include the substitution of the lysine residue at position 120 with a glutamic acid residue claim 55 , and wherein the protein has ethylenediamine-N claim 55 ,N′-disuccinate:ethylenediamine lyase activity that is heat resistant.61. The isolated gene of claim 60 , wherein the isoleucine residue at position 166 is substituted with a serine residue.62. The ...

SYSTEMS AND METHODS FOR BIOTRANSFORMATION OF CARBON DIOXIDE INTO HIGHER CARBON COMPOUNDS

Systems, compounds and methods for the conversion of C1 carbon compounds to higher carbon compounds useful for the generation of commodity compounds. 1. A method for converting formaldehyde to dihydroxyacetone , comprising contacting formaldehyde with a thiamine diphosphate cofactor and a polypeptide comprising a motif capable of associating with the thiamine diphosphate cofactor ,wherein the motif comprises a histidine residue in a first position, an asparagine or glutamine residue in a second position, and a glutamic acid or aspartic acid residue in a third position;wherein upon association with the thiamine diphosphate cofactor, the cofactor pyrimidine N1 is hydrogen bonded to the acid side chain of the glutamic acid residue or the acid side chain of the aspartic acid residue of the motif; andwherein the histidine N1 and the amide side chain of the asparagine residue or the amide side chain of the glutamine residue are capable of making a water-mediated hydrogen bond with the formaldehyde substrate and/or dihydroxyacetone product.2. The method of claim 1 , wherein the motif is incorporated in a benzaldehyde lyase polypeptide scaffold.3. The method of claim 2 , wherein the benzaldehyde lyase polypeptide scaffold comprises an amino acid sequence with at least 40% identity to the sequence set forth in SEQ ID NO:2 or SEQ ID NO:3.4. The method of claim 1 , wherein the polypeptide is engineered.5. The method of claim 4 , wherein the motif comprises at least one amino acid substitution corresponding to the substitutions selected from the group consisting of A394G claim 4 , A480W claim 4 , G419N claim 4 , A28S claim 4 , and A28I claim 4 , with reference to the amino acid sequence set forth in SEQ ID NO:2.6. The method of claim 5 , wherein the motif comprises the amino acid substitutions corresponding to A394G and A480W claim 5 , with reference to the amino acid sequence set forth in SEQ ID NO:2.7. The method of claim 4 , wherein the motif comprises at least three amino ...

PRODUCTION OF ISOPRENOIDS UNDER NEUTRAL pH CONDITIONS

Embodiments of the present disclosure relate to a process for producing isoprenoid precursor molecules and/or isoprenoids from a starch substrate by saccharification and/or fermentation. The saccharification is effectively catalyzed by a glucoamylase at a pH in the range of 5.0 to 8.0. At a pH of 6.0 or above, the glucoamylase possesses at least 50% activity relative to its maximum activity. The saccharification and fermentation may be performed as a simultaneous saccharification and fermentation (SSF) process. 1Humicola griseaTrichoderma reeseiRhizopus. A method for producing an isoprenoid precursor or isoprenoid comprising culturing a host cell , which comprises a heterologous nucleic acid encoding an polyprenyl pyrophosphate synthase polypeptide , and saccharifying and fermenting a starch substrate under simultaneous saccharification and fermentation (SSF) conditions in the presence of a glucoamylase , wherein the saccharification and fermentation are performed at pH 5.0 to 8.0 , wherein the glucoamylase possesses at least 50% activity at pH 6.0 or above relative to its maximum activity , wherein the glucoamylase is selected from the group consisting of a parent glucoamylase (HgGA) comprising SEQ ID NO: 3 , a parent glucoamylase (TrGA) comprising SEQ ID NO: 6 , a parent sp. glucoamylase (RhGA) comprising SEQ ID NO: 9 , and a variant thereof , and wherein the variant has at least 99% sequence identity to the parent glucoamylase.2. The method of claim 1 , wherein the isoprenoid is selected from group consisting of monoterpenes claim 1 , diterpenes claim 1 , triterpenes claim 1 , tetraterpenes claim 1 , sequiterpene claim 1 , and polyterpene.3. The method of claim 1 , wherein the isoprenoid is a sesquiterpene.4. The method of claim 1 , wherein the isoprenoid is selected from the group consisting of abietadiene claim 1 , amorphadiene claim 1 , carene claim 1 , α-farnesene claim 1 , β-farnesene claim 1 , farnesol claim 1 , geraniol claim 1 , geranylgeraniol claim 1 , ...

HEAT-STABLE CARBONIC ANHYDRASES AND THEIR USE

The present invention relates to use of heat-stable carbonic anhydrase in COextraction, e.g., from flue gas, natural gas or biogas. Furthermore, the invention relates to isolated polypeptides having carbonic anhydrase activity at elevated temperatures and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides. 1Bacillus halodurans. A method of using a heat-stable carbonic anhydrase for extraction of carbon dioxide from a carbon dioxide-containing medium , comprising contacting a carbon dioxide-containing medium with a heat stable carbonic anhydrase that maintains activity at temperatures above 45° C. for at least 15 minutes , wherein the heat stable carbonic anhydrase is an alpha-carbonic anhydrase which is at least 85% identical to the alpha-carbonic anhydrase from (SEQ ID NO: 16) , wherein carbon dioxide is extracted from the carbon dioxide-containing medium.2Bacillus halodurans. The method in accordance with claim 1 , wherein the carbonic anhydrase is the alpha-carbonic anhydrase from (SEQ ID NO: 16).3. The method of claim 1 , where the carbonic anhydrase maintains activity at temperatures above 45° C. for at least two hours.4. The method of claim 1 , where the carbonic anhydrase is used in a bioreactor.5. The method of claim 4 , wherein the bioreactor comprises a contained liquid membrane.6. The method of claim 5 , wherein the membrane liquid of the contained liquid membrane contains a bicarbonate buffer with a pH of at least 9.0.7. The method of claim 1 , wherein the carbon dioxide-containing medium is a carbonic dioxide-containing gas.8. The method of claim 1 , wherein the carbon dioxide-containing medium is a multiphase mixture.9. The method of claim 7 , where the carbonic dioxide-containing gas is emitted from combustion or fermentation.10. The method of claim 7 , where the carbonic dioxide-containing gas is a flue gas.11. The method of claim 7 , where ...

Protoilludene synthase

The present invention relates to an isolated, recombinant or synthetic polynucleotide encoding a polypeptide with protoilludene synthase activity and comprising a sequence selected from the group consisting of a) SEQ ID Nos. 1 or 14 of the attached sequence listing; b) a nucleic acid sequence complementary to SEQ ID Nos. 1 or 14; c) nucleic acid sequences which hybridize under stringent conditions to the nucleic acid sequences defined in a) and b) or their complementary strands, as well as to the polypeptide encoded by the isolated polynucleotide, as well as a method for the production of melleolides employing the polynucleotide or polypeptide of the invention.

NOVEL STRAINS OF MICROALGAE OF THE GENUS BOTRYOCOCCUS AND METHOD FOR THE CULTURE OF SAID MICROALGAE IN MIXOTROPHIC MODE

Novel strains of microalgae which belong to the genus and which can grow in a mixotrophic mode, and a cultivation method which includes providing light in the form of flashes for the production of lipids and hydrocarbons, in particular in the form of botryococcenes, which are useful in the production of biofuel. 1Botryococcus. Method for the culture of microalgae of the genus for the production of lipids or hydrocarbons , characterized in that it comprises the following steps:{'i': 'Botryococcus', 'sup': −2', '−1, 'the culture of one or more strains of microalgae of the genus in darkness in the presence of a supply of light that is discontinuous or variable over time, the intensity of which, in micromoles of photons, varies by an amplitude of more than 10 μmol. m. s, at a rate of at least once per hour;'}the maintenance of said culture over several generations in the presence of a carbon-containing substrate in the culture medium;the harvesting of the cells charged with hydrocarbons or lipids.2. Method according to claim 1 , characterized in that the supply of light is discontinuous.3. Method according to claim 1 , characterized in that the supply of light varies by more than 40 claim 1 , preferably claim 1 , more than 50 μmol. m. s.4. Method according to claim 1 , characterized in that it also comprises the recovery of the lipids or hydrocarbons contained in or excreted by the microalgae.5. Method according to claim 4 , characterized in that the hydrocarbons contained in or excreted by the microalgae comprise botryococcenes.6. Method according to claim 1 , characterized in that the culture medium is a minimum medium comprising a carbon-containing substrate.7. Method according to claim 1 , characterized in that the carbon-containing substrate comprises acetate claim 1 , glucose claim 1 , cellulose claim 1 , starch claim 1 , lactose claim 1 , saccharose or glycerol.8. Method according to claim 1 , characterized in that the supply of light is carried out in the form ...

Diterpene Synthases And Method For Producing Diterpenoids

Provided herein are diterpene synthases (diTPS) and methods for producing diterpenoids. Also provided herein are nucleic acid sequences encoding diTPS, diTPS amino acid sequences, diTPS proteins, vectors, cells, transgenic organisms, uses, compositions, methods, processes, and kits thereof. 1. An isolated nucleic acid molecule , comprising a sequence of nucleotides encoding a cis-abienol synthase (CAS) polypeptide selected from among:a) the polypeptide whose sequence is set forth in one of SEQ ID NOS:7, 50 and 55;b) an active fragment of the polypeptide of a); andc) a polypeptide having a sequence of amino acids that has at least 85% sequence identity with a polypeptide of a) or b),wherein the encoded polypeptide or active fragment catalyzes the formation of cis-abienol from geranylgeranyl diphosphate (GGPP).2. The isolated nucleic acid molecule of claim 1 , wherein the sequence of nucleotides encodes a polypeptide that comprises the sequence of amino acids set forth in one of SEQ ID NOS:7 claim 1 , 50 and 55 or an active fragment thereof.3. The isolated nucleic acid molecule of claim 1 , wherein the sequence of nucleotides encodes a polypeptide that consists of the sequence of amino acids set forth in one of SEQ ID NOS:7 claim 1 , 50 and 55 or an active fragment thereof.4. The isolated nucleic acid molecule of claim 1 , wherein the active fragment is a pseudomature form.5. The isolated nucleic acid molecule of claim 4 , wherein the sequence of nucleotides encodes a cis-abienol synthase (CAS) polypeptide whose sequence is set forth in SEQ ID NO:50 or 55 claim 4 , or encodes a polypeptide whose sequence has at least 85% sequence identity with SEQ ID NO:50 or 55.6. The isolated nucleic acid molecule of claim 1 , comprising a sequence of nucleotides selected from among:a) the nucleic acid molecule whose sequence is set forth in one of SEQ ID NOS:8, 54 and 56; andb) a nucleic acid molecule whose sequence of nucleotides has at least 85% sequence identity to the sequence ...

Filamentous Fungi and Methods for Producing Isoprenoids

The present invention relates to the production of a isoprenoid products from a lignocellulosic feedstock. Specifically at least triple mutant of filamentous fungi having the isoprenoid pathway results in production of isoprenoid products in commercial quantities. One embodiment of the invention relates to producing the isoprenoid products at the site of the lignocellulosic feedstock to reduce costs of shipping the feedstock. 1. A mutant isoprenoid producing filamentous fungus having the trichothecenes pathway comprising:a) a disrupted Tri5 gene, or a mutant Tri5 gene having low trichodiene synthase production; andb) a modified Tri6 gene, a modified Tri10 gene, or a modified gene encoding a terpene synthase, the gene modified to increase production of the gene product;wherein the mutant filamentous fungus produces at least 10% more isoprenoid product than the parent filamentous fungal cell when cultured under the same conditions.2. The mutant filamentous fungus according to wherein the modified gene has been modified to have inducible activity in producing a gene product.3. The mutant filamentous fungus according to wherein the modified gene has been modified to have constitutive activity in producing a gene product.4Acremonium, Aspergillus, Aureobasidium, Cryptococcus, Filibasidium, Fusariuni, Gibberella, Humicola, Magnaporthe, Mucor, Myceliophthora, Myrothecium, Neocallimastix, Neurospora, Paecilomyces, Penicillium, Piromyces, Schizophyllum, Stachybotrys, Talaromyces, Thermoascus, Thielavia, TolypocladiurnTrichoderma. The mutant filamentous fungus according to wherein the filamentous fungus is selected from the group consisting of claim 1 , or strain.5Fusarium. The mutant filamentous fungus according to wherein the filamentous fungus is a species.6FusariumFusarium sporotrichioides.. The mutant filamentous fungus according to wherein the species is7FusariumFusarium venenatum.. The mutant filamentous fungus according to wherein the species is8. The mutant ...

PRODUCTION OF ACETYL-COENZYME A DERIVED ISOPRENOIDS

Provided herein are compositions and methods for the heterologous production of acetyl-CoA-derived isoprenoids in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an acetaldehyde dehydrogenase, acetylating (ADA, E.C. 1.2.1.10) and an MEV pathway comprising an NADH-using HMG-CoA reductase. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an ADA and an MEV pathway comprising an acetoacetyl-CoA synthase. In some embodiments, the genetically modified host cell further comprises one or more heterologous nucleotide sequences encoding a phosphoketolase and a phosphotransacetylase. In some embodiments, the genetically modified host cell further comprises a functional disruption of the native PDH-bypass. The compositions and methods described herein provide an energy-efficient yet redox balanced route for the heterologous production of acetyl-CoA-derived isoprenoids. 155-. (canceled)56. A genetically modified yeast cell capable of producing an isoprenoid , the cell comprising:(a) a nucleic acid encoding an enzyme that condenses acetyl-CoA with malonyl-CoA to form acetoacetyl-CoA, and optionally one or more heterologous nucleic acids encoding one or more enzymes of a mevalonate (MEV) pathway for making isopentenyl pyrophosphate;(b) a heterologous nucleic acid encoding an acylating acetylaldehyde dehydrogenase (ADA, EC 1.2.1.10); and(c) a functional disruption of one or more enzymes of the native pyruvate dehydrogenase (PDH)-bypass selected from the group consisting of acetyl-CoA synthetase 1 (ACS1), acetyl-CoA synthetase 2 (ACS2), and aldehyde dehydrogenase 6 (ALD6),wherein the genetically modified yeast cell produces an increased amount of an isoprenoid compound compared to an yeast cell not comprising a nucleic acid encoding an enzyme that condenses acetyl-CoA with malonyl-CoA to form acetoacetyl-CoA.57. The genetically modified ...

LEUCINE BETA ROLL DOMAINS AND USES THEREOF

In one aspect, the invention relates to a peptide that forms a calcium-dependent hydrogel using a rationally engineered beta roll peptide. In the absence of calcium, the peptide is intrinsically disordered. Upon addition of calcium, the peptide forms a corkscrew-like structure. In one embodiment, one face of the beta roll is mutated to comprise leucine residues. In some embodiments, a leucine zipper forming helical domain to the engineered beta roll forms hydrogels by physical cross-linking in calcium rich environments. 1. A beta roll comprising a scaffold from the RTX domain of adenylate cyclase , wherein leucine mutations are introduced on the beta roll domain.2. The beta roll of claim 1 , wherein the beta roll domain comprises the amino acid sequence XXXXXXXXX(SEQ ID NO: 10) claim 1 , wherein{'sub': '1', '(a) Xis an amino acid selected from the group consisting of glycine, valine and serine;'}{'sub': '2', '(b) Xis an amino acid selected from the group consisting of glycine, serine, aspartic acid and leucine;'}{'sub': '3', '(c) Xis an amino acid selected from the group consisting of alanine, glutamic acid, glutamine, tyrosine and glycine;'}{'sub': '4', '(d) Xis an amino acid selected from the group consisting of glycine and arginine;'}{'sub': '5', '(e) Xis an amino acid selected from the group consisting of aspartic acid, alanine, asparagine, serine, and histidine;'}{'sub': '6', '(f) Xis an amino acid selected from the group consisting of aspartic acid and asparagine;'}{'sub': '7', '(g) Xis an amino acid selected from the group consisting of valine, leucine, and threonine;'}{'sub': '8', '(h) Xis an amino acid selected from the group consisting of leucine, isoleucine, and tyrosine; and'}{'sub': '9', '(i) Xis an amino acid selected from the group consisting of isoleucine, leucine, serine, and arginine.'}3. The beta roll of claim 2 , wherein the beta roll domain further comprises X claim 2 , wherein Xis appended adjacent to X claim 2 , and wherein{'sub': '10', '(a) ...

Method for biocatalytic production of nitriles from oximes and oxime dehydratases usable therein

The present invention relates to novel methods for biocatalytic production of nitriles from oximes using oxime dehydratases and novel mutants with oxime dehydratase activity and use thereof in a process for biocatalytic production of nitriles, such as in particular for the production of citral nitrile, neral nitrile, geranial nitrile or citronellyl nitrile from citral oxime, neral oxime, geranial oxime or citronellal oxime; and oxime dehydratases usable therefor, nucleotide sequences therefor and expression constructs or microorganisms comprising these.

Purification of Cystathionine Beta-Synthase

This invention provides chromatographic methods for the purification of a cystathionine β-Synthase (CBS) protein, particularly truncated variants thereof and compositions and pharmaceutical compositions prepared therefrom. 1. A method for purifying cystathionine β-Synthase (CBS) protein , wherein the CBS protein is a naturally occurring , chemically cleaved or genetically engineered truncated CBS protein , comprising the steps of:(a) providing a CBS-containing solution comprising one or a plurality of impurities; and(b) performing chromatographic separation of the CBS-containing solution using an ion exchange chromatography column and a metal affinity chromatography (IMAC) resin, wherein the impurities are removed thereby.2. The method of claim 1 , further comprising the step of performing chromatographic separation using a Hydrophobic Interaction Chromatography (HIC) column.3. The method of claim 1 , further comprising the step of performing chromatographic separation using a ceramic hydroxyapaptite resin.4. The method of claim 1 , wherein the ion exchange chromatography column is a weak anion exchanger.5. The method of claim 4 , wherein the weak anion exchanger is a DEAE-Sepharose FF column.6. The method of claim 1 , wherein the metal affinity chromatography (IMAC) resin is charged with a divalent metal cation.7. The method of claim 6 , wherein the divalent metal cation is nickel claim 6 , copper claim 6 , cobalt or zinc.8. The method of claim 7 , wherein the divalent metal ion is zinc.9. The method of claim 1 , further comprising eluting CBS from the metal affinity chromatography (IMAC) resin with an elution buffer comprising imidazole.10. The method of claim 1 , wherein the truncated CBS protein has an amino acid sequence identified by SEQ ID NO: 3.11. The method of claim 1 , wherein the CBS-containing solution is a clarified CBS solution.12. The method of claim 1 , wherein the CBS is produced in a recombinant cell.13. The method of claim 12 , wherein the ...

Production of Isoprenoids

Methods for producing an isoprenoid are provided. A plurality of bacterial or fungal host cells is obtained. These cells comprise a heterologous nucleic acid encoding one or more enzymes of a mevalonate pathway for making isopentenyl pyrophosphate. Expression of the one or more enzymes is under control of at least one heterologous transcriptional regulator. The mevalonate pathway comprises (i) an enzyme that condenses acetoacetyl-CoA with acetyl-CoA to form HMG-CoA, (ii) an enzyme that converts HMG-CoA to mevalonate, (iii) an enzyme that phosphorylates mevalonate to mevalonate 5-phosphate, (iv) an enzyme that converts mevalonate 5-phosphate to mevalonate 5-pyrophosphate, and (v) an enzyme that converts mevalonate 5-pyrophosphate to isopentenyl pyrophosphate. The host cells are cultured in a medium under conditions that are suboptimal as compared to conditions. Temperature is maintained at a level below that which would provide for a maximum specific growth rate for the host cells. 1. A method for producing an isoprenoid comprising:(a) obtaining a plurality of bacterial or fungal host cells that comprise a heterologous nucleic acid encoding one or more enzymes of a mevalonate pathway for making isopentenyl pyrophosphate, wherein expression of the one or more enzymes is under control of at least one heterologous transcriptional regulator, wherein said mevalonate pathway comprises (i) an enzyme that condenses acetoacetyl-CoA with acetyl-CoA to form HMG-CoA; (ii) an enzyme that converts HMG-CoA to mevalonate; (iii) an enzyme that phosphorylates mevalonate to mevalonate 5-phosphate; (iv) an enzyme that converts mevalonate 5-phosphate to mevalonate 5-pyrophosphate; and (v) an enzyme that converts mevalonate 5-pyrophosphate to isopentenyl pyrophosphate; and(b) culturing the host cells in a medium wherein temperature is maintained at a level below that which would provide for a maximum specific growth rate for the plurality of bacterial or fungal host cells.2. (canceled)3. ...

Three-dimensional structure of isoprene synthase and its use thereof for generating variants

The present invention provides a three-dimensional structures of P. tremuloides isoprene synthase and P. alba isoprene synthase. The invention also provides methods of using the three-dimensional structure to design isoprene synthases with improved activity for increased isoprene production in microbial host cells. Biosynthetically produced isoprene of the present invention finds use in the manufacture of rubber and elastomers.

NOVEL ALDOLASE AND PRODUCTION PROCESS OF SUBSTITUTED ALPHA-KETO ACIDS

4-(Indol-3-ylmethyl)-4-hydroxy-2-oxoglutarate, which is useful as an intermediate in the synthesis of monatin, may be synthesized from indole pyruvic acid and pyruvic acid (and/or oxaloacetic acid) by using a novel aldolase derived from the genus , or 1. An isolated polynucleotide of following (a) or (b):(e) a polynucleotide having the nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15; or(f) a polynucleotide that hybridizes under stringent conditions with a polynucleotide having the nucleotide sequence complementary to the full-length nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15, and encodes a protein having aldolase activity, wherein said stringent conditions comprise a wash step in 0.1×SSC and 0.1% SDS at 65° C.2. An isolated polynucleotide of following (c) or (d):(g) a polynucleotide that encodes a protein comprising the amino acid sequence of SEQ ID NO: 16; or(h) a polynucleotide that encodes a protein having an amino acid sequence that contains a substitution, deletion, insertion, or addition of one to ten amino acid residues in the amino acid sequence of SEQ ID NO: 16, and has aldolase activity.3. A recombinant DNA produced by ligating into a vector DNA a polynucleotide selected from the group consisting of:(e) a polynucleotide having the nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15;(f) a polynucleotide that hybridizes under stringent conditions with a polynucleotide having the nucleotide sequence complementary to the full-length nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15, and encodes a protein having aldolase activity, wherein said stringent conditions comprise a wash step in 0.1×SSC and 0.1% SDS at 65° C.;(g) a polynucleotide that encodes a protein comprising the amino acid sequence of SEQ ID NO: 16; and(h) a ...

ISOPRENE SYNTHASE VARIANTS FOR IMPROVED MICROBIAL PRODUCTION OF ISOPRENE

The present invention provides methods and compositions comprising at least one isoprene synthase enzyme with improved catalytic activity and/or solubility. In particular, the present invention provides variant plant isoprene synthases for increased isoprene production in microbial host cells. Biosynthetically produced isoprene of the present invention finds use in the manufacture of rubber and elastomers. 156-. (canceled)57. An isolated host cell comprising a heterologous polynucleotide sequence encoding an isoprene synthase variant in operable combination with a promoter , wherein said isoprene synthase variant comprises one or more amino acid substitution(s) at one or more amino acid residues corresponding to a poplar isoprene synthase having the sequence of SEQ ID NO: 120 , wherein said substitution(s) are selected from the group consisting of V10M , F12S , T15A , E18G , V58I , V58F , L70Q , L70R , L70V , L70T , T71P , V79L , E89D , G94A , S119F , F120L , G127R , E175V , T212I , S257A , R262G , A266G , F280L , N297K , F305L , L319M , E323K , A328T , D342E , A359T , K366N , E368D , L374M , S396T , V4185 , K438N , H440R , T442A , I449V , A469S , K500R , K505Q , G507S , S509N , F511Y , and N532K; andwherein the variant is capable of more effectively converting dimethylallyl diphosphate (DMAPP) to isoprene, as compared to an isoprene synthase variant without a substitution.58. The host cell of wherein at least one amino acid substitution is a L70R substitution.59. The host cell of wherein at least one amino acid substitution is a G507S substitution.60. The host cell of wherein the variant comprises one of more amino acid substitutions selected from the group consisting of G127R/F511Y claim 57 , L70Q/G94A/R262G/F305L claim 57 , F12S/T15A/E18G/N297K claim 57 , S396T/T442I claim 57 , V10M/E323K claim 57 , F120L/A266G claim 57 , K438N/K500R claim 57 , V79L/S509N claim 57 , E175V/S257A/E368D/A469S claim 57 , T71P/L374M claim 57 , F280L/H440R claim 57 , E89D/H440R claim ...

ENZYMES THAT SYNTHESIZE ZINGIBERENE

The invention relates to nucleic acids encoding a zingiberene synthase that enables host cells and plants to make zingiberene that is useful in fragrances and for repelling or killing insects. The invention also relates to isolated zingiberene synthases and to methods for making zingiberenes. 1. An isolated nucleic acid encoding a zingiberene synthase with at least 95% sequence identity to amino acid SEQ ID NO: 2 , 4 , 6 , 8 , 11 , 12 , 14 , 16 , 18 , or a combination thereof.2. The isolated nucleic acid of claim 1 , wherein the nucleic acid comprises a sequence with at least 86% sequence identity to any of nucleotide sequences SEQ ID NO:1 claim 1 , 3 claim 1 , 5 claim 1 , 7 claim 1 , 13 claim 1 , 15 claim 1 , 17 claim 1 , 19 claim 1 , or a combination thereof.3. An expression cassette comprising the nucleic acid of operably linked to a promoter functional in a host cell.4. A host cell comprising the nucleic acid of .5. The host cell of claim 4 , further comprising a promoter operably linked to the nucleic acid claim 4 , wherein the promoter is functional in the host cell.6. The host cell of claim 5 , wherein the host cell is a plant cell.7. The host cell of claim 5 , wherein the host cell is a microorganism.8. A plant tissue comprising the nucleic acid of .9. A plant tissue comprising the host cell of .10. A plant comprising the nucleic acid of .11. A plant comprising the plant tissue of .12. A method of making a zingiberene comprising:{'claim-ref': {'@idref': 'CLM-00004', 'claim 4'}, 'a) culturing the host cell of under conditions sufficient for expression of the zingiberene synthase; and'}b) providing the host cell with a substrate for the zingiberene synthase to make the sesquiterpene.13. The method of claim 12 , wherein the substrate is 2Z claim 12 ,6Z-farnesyl diphosphate.14. The method of claim 12 , wherein the host cell is a bacterial or yeast cell.15E. coli. The method of claim 14 , wherein the bacterial cell is an cell.16. An isolated zingiberene synthase ...

1-deoxy-d-xylulose 5-phosphate synthase alleles responsible for enhanced terpene biosynthesis

A method of enhancement of the 1-deoxy-D-xylulose 5-phosphate synthase (DXS) activity of plants or bacteria to increase terpenes production in cells, an enhanced DXS sequence likely to be obtained by this method, a method of enhancement of production of terpenes in a host cell containing the enhanced DXS enzyme, and transgenic bacterium or plants that express this polypeptide are described.

INCREASED ISOPRENE PRODUCTION USING THE ARCHAEAL LOWER MEVALONATE PATHWAY

The invention features methods for producing isoprene from cultured cells using a feedback-resistant mevalonate kinase polypeptide, such as an archaeal mevalonate kinase polypeptide. The resulting isoprene compositions may have increased yields and/or purity of isoprene. 113-. (canceled)14. A method of producing isoprene , the method comprising(a) culturing cells comprising (i) one or more heterologous nucleic acids encoding feedback-resistant mevalonate kinase polypeptides, (ii) one or more heterologous nucleic acids encoding an isoprene synthase polypeptide or one or more additional copies of an endogenous nucleic acid encoding an isoprene synthase polypeptide, and (iii) one or more nucleic acids encoding a mevalonate (MVA) pathway polypeptide under suitable culture conditions for the production of isoprene, and(b) producing isoprene.15. (canceled)16. The method of claim 14 , wherein the cells further comprise (iv) one or more nucleic acids encoding a DXP pathway enzyme.17. (canceled)18. The method of claim 14 , wherein the feedback-resistant mevalonate kinase is archaeal mevalonate kinase.19M. mazei. The method of claim 18 , wherein the archaeal mevalonate kinase polypeptide is mevalonate kinase.20. The method of claim 14 , wherein the cells in culture produce greater than about 400 nmole/g/hr of isoprene.21. The method of claim 14 , further comprising recovering the isoprene.22. A method of increasing the rate or flux of production of 3 claim 14 ,3-dimethylallyl diphosphate (DMAPP) claim 14 , isopentenyl diphosphate (IPP) claim 14 , or a product derived from 3 claim 14 ,3-dimethylallyl diphosphate (DMAPP) or isopentenyl diphosphate (IPP) comprising:(a) culturing cells comprising (i) one or more heterologous nucleic acids encoding feedback-resistant mevalonate kinase polypeptides, and (ii) one or more nucleic acids encoding a mevalonate (MVA) pathway polypeptide wherein the cells are cultured under suitable culture conditions for increasing the rate or flux of ...

DIRECT STARCH TO FERMENTABLE SUGAR AS FEEDSTOCK FOR THE PRODUCTION OF ISOPRENE, ISOPRENOID PRECURSOR MOLECULES, AND/OR ISOPRENOIDS

Provided herein are compositions and methods related to the direct conversion of the starch in a ground or fractionated grain into a fermentable sugar feedstock capable of serving as a carbon source for the industrial production of one or more products by a fermenting organism, such as isoprene, isoprenoid precursor molecules, and/or isoprenoids. Such conversions may be performed at temperatures at or below the initial gelatinization temperature of the starch present in the grain and may utilize one or more isolatable endogenous enzymes present in certain unrefined grains. 1. A method for the production of isoprene by recombinant host cells in culture , wherein the host cells comprise one or more heterologous nucleic acids encoding an isoprene synthase polypeptide and one or more mevalonate (MVA) pathway polypeptides , the method comprising:a. inactivating endogenous enzyme activity in a whole or fractionated grain;b. treating the whole or fractionated grain with a starch solubilizing alpha amylase and a glucoamylase, wherein the treatment is at a temperature at or below the initial gelatinization temperature of the starch in the grain, wherein the concentration of the alpha amylase is between about 5 to 20 AAU/gds, and wherein the treatment produces a fermentable sugar feedstock;c. culturing the recombinant host cells in culture media comprising the fermentable sugar feedstock; andd. producing isoprene.2. The method of claim 1 , wherein the treatment is at a temperature of about 0 to about 30° C. below the initial gelatinization temperature of the starch in the grain.3. The method of claim 1 , wherein the concentration of alpha amylase is about 6 AAU/g ds to about 10 AAU/g ds.4. The method of claim 1 , wherein the isoprene synthase polypeptide is a plant isoprene synthase polypeptide.5. The method of claim 4 , wherein the plant isoprene synthase polypeptide is a kudzu isoprene synthase polypeptide.6PuerariaPopulusPopulus alba×Populus tremula.. The method of claim 1 ...

REDUCTION OF CARBON DIOXIDE EMISSION DURING ISOPRENE PRODUCTION BY FERMENTATION

The present invention provides methods for increasing the amount of isoprene produced by cultured cells with only a minimal increase in carbon dioxide emitted, thereby resulting in process having a greater yield of isoprene relative to carbon dioxide. In addition, the present invention provides compositions that include the cultured cells or isoprene produced there from. 1: Cells comprising a heterologous nucleic acid that encodes an isoprene synthase polypeptide in operable combination with a promoter wherein the cells are in culture medium comprising one or more uncommon carbon sources and wherein the ratio of carbon dioxide emitted per isoprene produced in off gas from the cultured cells is less than 500.2: The cells of claim 1 , wherein the uncommon carbon source is selected from the group consisting of claim 1 , glycerol claim 1 , glycerine claim 1 , dihydroxyacetone claim 1 , one-carbon source claim 1 , oil claim 1 , animal fat claim 1 , animal oil claim 1 , fatty acid claim 1 , lipid claim 1 , phospholipid claim 1 , glycerolipid claim 1 , monoglyceride claim 1 , diglyceride claim 1 , triglyceride claim 1 , microbial polypeptide claim 1 , plant protein polypeptide claim 1 , algae claim 1 , and algae components.3: The cells of claim 2 , wherein the uncommon carbon source is selected from the group consisting of glycerol claim 2 , glycerine claim 2 , cell mass claim 2 , protein claim 2 , alcohol claim 2 , and plant-derived oil.4: The cells of claim 1 , wherein the cells further comprise one or more heterologous nucleic acids that encode for an MVA pathway enzyme or a DXP pathway enzyme.5: The cells of claim 4 , wherein the MVA pathway enzyme is mevalonate kinase.6M. mazeiLactobacillusLactobacillus sakeiSaccharomyces cerevisiaeStreptococcusStreptococcus pneumoniaeStreptomycesStreptomyces: The cells of claim 5 , wherein the mevalonate kinase is selected from the group consisting of feedback-resistant mevalonate kinase claim 5 , archaeal mevalonate kinase claim 5 , ...

ISOPRENE SYNTHASE VARIANTS WITH IMPROVED SOLUBILITY FOR PRODUCTION OF ISOPRENE

The present invention provides methods and compositions of variant polypeptides having isoprene synthase activity with improved solubility. In particular, the present invention provides isoprene synthase variant for increased isoprene production in recombinant host cells.

COMPOSITIONS AND METHODS OF PGL FOR THE INCREASED PRODUCTION OF ISOPRENE

Provided herein are improved compositions and methods for the increased production of isoprene. Also provided herein are improved compositions and methods for the increased production of heterologous polypeptides capable of biological activity. 1Escherichia coliE. coliE. coli. A method of producing isoprene , the method comprising: (a) culturing a composition comprising a recombinant cell of an () strain , or progeny thereof , under suitable culture conditions for the production of isoprene , wherein the cell comprises (i) one or more copies of a heterologous nucleic acid(s) encoding a 6-phosphogluconolactonase (PGL) polypeptide , wherein the nucleic acid is integrated in the chromosome and (ii) one or more heterologous nucleic acid(s) encoding isoprene synthase ,{'i': 'E. coli', 'wherein prior to the integration, the does not contain nucleic acids(s) encoding a PGL polypeptide, and wherein the resulting cell produces isoprene at a greater titer than that of the same cells that do not comprise (i); and'}(b) producing isoprene.2. The method of comprising further recovering the isoprene.3. The method of claim 1 , wherein the recombinant cell has a specific productivity greater than about 15 mg/OD/hr of isoprene.4E. coli. The method of claim 1 , wherein the PGL polypeptide is an PGL polypeptide.5. The method of claim 4 , wherein nucleic acids encoding the PGL polypeptide is part of a 17 claim 4 ,257 base pair piece as shown in .6. The method of claim 1 , wherein the cell produces isoprene at a higher specific productivity than that of the same cells that do not contain (i).7. The method of claim 1 , wherein the specific productivity of isoprene by the cell is at least 15 mg/OD/hr.8E. coliE. coli. The method of claim 4 , wherein the nucleic acid encoding PGL polypeptide is from strain K12 MG1655 or a derivative of strain K12 MG1655.9E. coli. The method of claim 1 , wherein the cell is of strain B.10E. coli. The method of claim 9 , wherein the cell is of strain BL21.11E. ...

EXPRESSION CONSTRUCTS AND USES THEREOF IN THE PRODUCTION OF TERPENOIDS IN YEAST

A method of producing at least one terpene in a yeast cell is disclosed. The method comprises exogenously expressing within the mitochondria of the yeast cell or directing localization thereto a terpene synthase. 1. A method of producing at least one terpene in a yeast cell , the method comprising exogenously expressing within the mitochondria of the yeast cell or directing localization thereto a terpene synthase , thereby producing the at least one terpene in the yeast cell.2. The method of claim 1 , wherein said terpene synthase is translationally fused to a mitochondrial localization signal (MLS) peptide.3. The method of claim 1 , further comprising exogenously expressing within the yeast cell an enzyme in a terpenoid/sterol pathway which catalyzes formation of a farnesyl diphosphate (FDP).4. The method of claim 3 , wherein said exogenously expressing within the yeast cell said enzyme in said terpenoid/sterol pathway which catalyzes formation of said farnesyl diphosphate is effected in the mitochondria of the yeast cell or by directing localization of said enzyme to said mitochondria of the yeast cell.5. The method of claim 4 , wherein said enzyme in said terpenoid/sterol pathway is translationally fused to a mitochondrial localization signal (MLS) peptide.6. The method of claim 1 , further comprising exogenously expressing within the yeast cell a mutated form of yeast 3-hydroxy-3-methylglutaryl-coenzyme A reductase (tHMG).7. The method of claim 1 , further comprising exogenously expressing within the yeast cell a terpene synthase claim 1 , wherein said terpene synthase is not expressed in claim 1 , or directed to said mitochondria.8. The method of claim 1 , wherein said terpene synthase is selected from the group consisting of a valencene synthase claim 1 , a linalool synthase claim 1 , a phytoene synthase claim 1 , an amorphadiene synthase claim 1 , a limonene synthase and a taxadiene synthase.9. The method of claim 3 , wherein said enzyme in said terpenoid/ ...

MICROORGANISMS AND PROCESSES FOR THE PRODUCTION OF ISOPRENE

The present invention provides a novel biosynthetic pathway for the production of isoprene from 3-methyl-2-buten-1-ol or 2-methyl-3-buten-2-ol. Further embodiments provide non-naturally occurring microorganism that have been modified to produce isoprene from 3-methyl-2-buten-1-ol or 2-methyl-3-buten-2-ol and methods of producing isoprene using said microorganism. 1. A non-naturally occurring microbial organism comprising an isoprene biosynthetic pathway for the conversion of dimethylallyl diphosphate to isoprene , wherein the isoprene biosynthetic pathway comprises an exogenous nucleic acid encoding at least one enzyme of an isoprene biosynthetic pathway selected from: 2-methyl-3-buten-2-ol synthase; 2-methyl-3-buten-2-ol dehydratase; and any combination thereof; and wherein the isoprene biosynthetic pathway is expressed at a sufficient level to produce isoprene.2. The non-naturally occurring microbial organism of claim 1 , wherein the organism overexpresses one or more endogenous or exogenous genes encoding at least one enzyme selected from: an enzyme of the methylerythritol phosphate pathway or an enzyme of the mevalonate pathway.3. The non-naturally occurring microbial organism of claim 2 , wherein the dimethylallyl diphosphate available for conversion to isoprene is increased.4. The non-naturally occurring microbial organism of claim 1 , wherein the 2-methyl-3-buten-2-ol dehydratase is a linalool dehydratase-isomerase.5Castellaniella defragrans.. The non-naturally occurring microbial organism of claim 4 , wherein the 2-methyl-3-buten-2-ol dehydratase is a linalool dehydratase-isomerase derived from6. The non-naturally occurring microbial organism of claim 1 , further comprising one or more endogenous or exogenous genes encoding at least one enzyme of the methylerythritol phosphate pathway selected from: 1-deoxy-D-xylulose-5-phosphate synthase claim 1 , 1-deoxy-D-xylulose-5-phosphate reductoisomerase claim 1 , 4-diphosphocytidyl-2-C-methyl-D-erythritol synthase ...

MICROORGANISMS AND PROCESSES FOR THE PRODUCTION OF ISOPRENE

The present invention provides a novel biosynthetic pathway for the production of isoprene from 3-methyl-2-buten-1-ol or 2-methyl-3-buten-2-ol. Further embodiments provide non-naturally occurring microorganism that have been modified to produce isoprene from 3-methyl-2-buten-1-ol or 2-methyl-3-buten-2-ol and methods of producing isoprene using said microorganism. 1. A non-naturally occurring microbial organism comprising an isoprene biosynthetic pathway for conversion of dimethylallyl diphosphate to isoprene , wherein the isoprene biosynthetic pathway comprises an exogenous nucleic acid encoding at least one enzyme of an isoprene biosynthetic pathway selected from: 3-methyl-2-buten-1-ol synthase; 2-methyl-3-buten-2-ol isomerase; 2-methyl-3-buten-2-ol dehydratase; and any combination thereof; and wherein the isoprene biosynthetic pathway is expressed at a sufficient level to produce isoprene.2. The non-naturally occurring microbial organism of claim 1 , wherein the organism overexpresses one or more endogenous or exogenous genes encoding at least one enzyme selected from: an enzyme of the methylerythritol phosphate pathway or an enzyme of the mevalonate pathway.3. The non-naturally occurring microbial organism of claim 2 , wherein the dimethylallyl diphosphate available for conversion to isoprene is increased.4. The non-naturally occurring microbial organism of claim 1 , wherein the 2-methyl-3-buten-2-ol dehydratase is a bi-functional enzyme further comprising 2-methyl-3-buten-2-ol isomerase activity.5. The non-naturally occurring microbial organism of claim 4 , wherein the 2-methyl-3-buten-2-ol dehydratase is a linalool dehydratase-isomerase.6Castellaniella defragrans.. The non-naturally occurring microbial organism of claim 5 , wherein the 2-methyl-3-buten-2-ol dehydratase is a linalool dehydratase-isomerase derived from7. The non-naturally occurring microbial organism of claim 1 , wherein the 3-methyl-2-buten-1-ol synthase is a phosphatase.8Bacillus ...

CONVERSION OF PRENYL DERIVATIVES TO ISOPRENE

The present invention provides methods for producing derivatives from cultured cells. In addition, the present invention provides methods for conversion of prenyl derivatives, obtained from biological or petrochemical sources, to isoprene by employing chemical or biological catalysts. The present invention also provides compositions that include the cultured cells or isoprene or prenyl derivatives produced there from. 2. The method of claim 1 , wherein the prenyl derivative is prenyl alcohol.3. The method of claim 1 , wherein the prenyl derivative is recovered from a fermentation of the cultured cells using one or more processes selected from the group consisting of distillation claim 1 , gas-stripping claim 1 , two-phase recovery claim 1 , and pervaporation.4. The method of claim 2 , wherein the process for recovering prenyl alcohol from a fermentation of the cultured cells comprises gas-stripping or two-phase recovery.5. The method of claim 1 , wherein the prenyl derivative comprises one or more of prenol claim 1 , isoprenol claim 1 , 3-methyl-3-buten-1-yl acetate claim 1 , and 3-methyl-2-buten-1-yl acetate.6. The method of claim 1 , wherein the cells further comprise one or more of an isopentenyl-diphosphate-delta-isomerase (IDI) polypeptide claim 1 , one or more mevalonic acid (MVA) pathway polypeptides claim 1 , and/or one or more deoxyxylulose-5-phosphate (DXP) pathway polypeptides.7. The method of claim 1 , wherein the cells further comprise one or more lower MVA pathway polypeptides.8. The method of claim 1 , wherein the one or more phosphatases of enzyme class 3.6.1 claim 1 , 3.1.7 or 3.1.3 are one or more phosphatases selected from the group consisting of an allyl diphosphatase claim 1 , an ADP-ribose pyrophosphatase claim 1 , an ADP-sugar phosphorylase claim 1 , a nucleoside triphosphate pyrophosphatase claim 1 , a FAD pyrophosphatase claim 1 , a monoterpenyl pyrophosphastase claim 1 , an alkaline phosphatase claim 1 , and an acid phosphatase.9. The ...

LEGUME ISOPRENE SYNTHASE FOR PRODUCTION OF ISOPRENE

The present invention provides methods and compositions of polypeptides having isoprene synthase activity with improved performance characteristics. In particular, the present invention provides legume isoprene synthases for increased isoprene production in recombinant host cells. 1P. montana.. An isolated polypeptide from a legume having isoprene synthase activity , wherein said polypeptide comprises at least 40% sequence identity to SEQ ID NO: 1 , wherein said polypeptide has one or more amino acid residue(s) corresponding to one or more amino acid residue(s) corresponding to SEQ ID NO:1 , wherein said one or more amino acid residue(s) are selected from the group consisting of F287 , G397 , N438 , E451 , and Y514 , and wherein said isolated polypeptide is not from2ArachisMucanaCajanusGlycineLotusMedicago. The isolated polypeptide of claim 1 , wherein the isolated polypeptide is an isoprene synthase selected from the group consisting of sp. claim 1 , sp. claim 1 , sp. claim 1 , sp. claim 1 , sp. claim 1 , and sp.3A. hypogaea, M. pruriens, C. cajans, G. max, G. soja, L. japonicusM. truncatula.. The isolated polypeptide of claim 2 , wherein the isolated polypeptide is an isoprene synthase selected from the group consisting of claim 2 , and4A. hypogaea. The isolated polypeptide of claim 2 , wherein the isolated polypeptide is an isoprene synthase.5M. pruriens. The isolated polypeptide of claim 2 , wherein the isolated polypeptide is an isoprene synthase.6. The isolated polypeptide of claim 1 , wherein said isolated polypeptide has a reduced substrate inhibition as compared to a poplar isoprene synthase.7. The isolated polypeptide of claim 1 , wherein said isolated polypeptide has increased isoprene synthase activity as compared to a poplar isoprene synthase.8. The isolated polypeptide of claim 7 , wherein increased isoprene synthase activity is indicated by a host cell comprising the isoprene synthase displaying improved growth in the presence of mevalonic acid ...

IDENTIFICATION OF ISOPRENE SYNTHASE VARIANTS WITH IMPROVED PROPERTIES FOR THE PRODUCTION OF ISOPRENE

The invention provides for compositions and methods for producing isoprene using isoprene synthase variants with improved properties. 1. An isolated polypeptide having isoprene synthase activity , wherein the polypeptide comprises one or more amino acid substitution(s) at one or more residues corresponding to SEQ ID NO:1 selected from the group consisting of: X323F , X118E , X36W , X22K , X228Y , X448L , X488F , X467H , X443S , X331P , X453I , X71K , X71L , X448I , X71M , X392Y , X448V , X282H , X383Y , X323Y , X511Y , X448E , X376M , X488L , X120E , X461A , X414I , X282W , X071K , X493E , X392S , X448Q , X282Y , X537N , X447Y , X240C , X443Q , X538R , and X510C , and wherein the polypeptide has improved Kcat compared to a parent polypeptide which does not comprise said one or more amino acid substitution(s).2. The polypeptide of wherein the amino acid substitution is selected from the group consisting of: D323F claim 1 , A118E claim 1 , K36W claim 1 , S22K claim 1 , M228Y claim 1 , A448L claim 1 , E488F claim 1 , E467H claim 1 , A443S claim 1 , C331P claim 1 , A453I claim 1 , R71K claim 1 , R71L claim 1 , A448I claim 1 , R71M claim 1 , W392Y claim 1 , A448V claim 1 , S282H claim 1 , T383Y claim 1 , D323Y claim 1 , H511Y claim 1 , A448E claim 1 , L376M claim 1 , E488L claim 1 , S120E claim 1 , R461A claim 1 , K414I claim 1 , S282W claim 1 , R071K claim 1 , S493E claim 1 , W392S claim 1 , A448Q claim 1 , S282Y claim 1 , E537N claim 1 , I447Y claim 1 , T240C claim 1 , A443Q claim 1 , P538R claim 1 , and S510C.36-. (canceled)7. An isolated polypeptide having isoprene synthase activity claim 1 , wherein the polypeptide comprises one or more amino acid substitution(s) at one or more residues corresponding to SEQ ID NO:1 selected from the group consisting of: X22K claim 1 , X348F claim 1 , X392V claim 1 , X392F claim 1 , X488C claim 1 , X22R claim 1 , X71V claim 1 , X443R claim 1 , X234R claim 1 , X453V claim 1 , X437Y claim 1 , X392C claim 1 , X463F claim 1 , X538K claim ...

Human Cystathionine Beta-Synthase Variants and Methods of Production Thereof

Human cystathionine β-synthase variants are disclosed, as well as a method to produce recombinant human cystathionine β-synthase and variants thereof. More particularly, the role of both the N-terminal and C-terminal regions of human CBS has been studied, and a variety of truncation mutants and modified CBS homologues are described. In addition, a method to express and purify recombinant human cystathionine β-synthase (CBS) and variants thereof which have only one or two additional amino acid residues at the N-terminus are described. 1. An isolated human cystathionine β-synthase (CBS) protein having CBS biological activity , said protein having: an amino acid sequence having an amino terminal deletion or a carboxyl terminal deletion or both an amino terminal and carboxyl terminal deletion and spanning from a starting position of one of amino acid residues from about 1-39 or 2-39 of SEQ ID NO: 2 to an ending position of one of amino acid residue 551 , or amino acid residue 413 , or an amino acid residue from about 397-551 or 414-551 of SEQ ID NO: 2; or a homologue thereof comprising an amino acid sequence that is at least about 70%-95% identical to said amino acid sequence , wherein said isolated human cystathionine β-synthase variant catalyzes the formation of cystathionine.28-. (canceled)9. The isolated human cystathionine β-synthase variant of claim 1 , wherein said ending position is amino acid residue 551 of SEQ ID NO: 2.10. The isolated human cystathionine β-synthase variant of claim 1 , wherein said ending position is amino acid residue 413 of SEQ ID NO: 2.11. The isolated human cystathionine β-synthase variant of claim 1 , wherein said ending position is one of amino acid residues from about 397-551 of SEQ ID NO: 2.12. The isolated human cystathionine β-synthase variant of claim 1 , wherein said ending position is one of amino acid residues from about 414-551 of SEQ ID NO: 2.1321-. (canceled)22. The isolated human cystathionine β-synthase protein of claim 1 , ...

ENOLASE PEPTIDE CONJUGATE VACCINES AGAINST STAPHYLOCOCCUS AUREUS

The present invention relates to peptides of the enolase protein from as well as nucleic acid and nucleic acid sequence homologues encoding the peptides. The present invention also relates to a composition, particularly a vaccine, comprising one or more of the enolase peptides described herein or a fragment, derivative or variant thereof capable of generating an immune response that induces a protective antibody response or opsonophagocytic activity of human neutrophils for . The present invention also encompasses methods of treating and/or reducing the likelihood of a infection by administering a composition of the invention. 1. An isolated peptide consisting of a sequence of amino acids selected from the group consisting of: SEQ ID NO: 1 , 2 , 5 and 6 , or a variant or derivative.2. The isolated peptide of claim 1 , wherein said peptide comprises a derivative of the amino acid sequence set forth in SEQ ID NO: 1 claim 1 , 2 claim 1 , 5 or 6.3. The isolated peptide of claim 1 , wherein said peptide comprises a variant of the amino acid sequence set forth in SEQ ID NO: 1 claim 1 , 2 claim 1 , 5 or 6.4. The isolated peptide of claim 2 , wherein said derivative comprises one or more additional regions or moieties covalently joined to said amino acid sequence claim 2 , wherein each region or moiety is independently selected from a region or moiety having at least one of the following properties: enhances the immune response claim 2 , facilitates purification claim 2 , or facilitates polypeptide stability.5. The isolated peptide of claim 4 , wherein said derivative consists of an amino acid sequence as set forth in SEQ ID NO:1 claim 4 , 2 claim 4 , 5 claim 4 , or 6 with an N-terminal methionine.6. The isolated peptide of claim 1 , wherein said peptide consists essentially of the amino acid sequence set forth in SEQ ID NO: 1 claim 1 , 2 claim 1 , 5 claim 1 , or 6.7S. aureus. A composition able to induce a protective immune response in a patient against infection ...

Terpene Synthases and Methods of Using the Same

Disclosed are isolated nucleic acid molecules from that encode a terpene synthase protein at least 80% identical to a protein encoded by the nucleic acid sequence according to any of SEQ ID NOs: 1-47 or a degenerate variant thereof or a functional fragment thereof. Isolated terpene synthase proteins from are also disclosed. Host cells transformed with the terpene synthase nucleic acids are also disclosed, for example cells of a single cell organism, such as bacteria and yeast, or multicellular organism, such as a plant. The host cells can be prokaryotic cells or eukaryotic cells. Transgenic plants, or any part thereof, stably transformed with terpene synthase nucleic acids are also disclosed In some examples the transgenic plant is a dicotyledon or a monocotyledon. A method is disclosed for producing a transgenic plant, as is a method for producing terpenes. 1. An isolated nucleic acid molecule , comprising a nucleic acid sequence that encodes a terpene synthase protein at least 80% identical to a protein encoded by the nucleic acid sequence according to any of SEQ ID NOs: 1-47 or a degenerate variant thereof or a functional fragment thereof.2. The isolated nucleic acid molecule of claim 1 , further comprising a promoter operably linked to the nucleic acid sequence that encodes the terpene synthase protein.3. The isolated nucleic acid molecule of claim 1 , wherein the isolated nucleic acid comprises the cDNA set forth as anyone of SEQ ID NOs: 1-47 claim 1 , or a degenerate variant thereof.4. A construct comprising isolated nucleic acid molecule of any one of .5. The construct of claim 4 , wherein the construct confers an agronomic trait to a plant in which it is expressed.6. The construct of claim 5 , wherein the agronomic trait comprises terpenoid production.7. An expression vector comprising the nucleic acid molecule of .8. A host cell transformed with the vector of .9. The host cell of claim 8 , where the cell comprises a prokaryotic cell or a eukaryotic cell.10. ...

CYAA-CARRIED POLYPEPTIDE(S) AND USE TO INDUCE BOTH THERAPEUTIC AND PROPHYLACTIC IMMUNE RESPONSES

The invention is directed to means, based on CyaA-carried polypeptide(s), for use in the immunotherapeutic treatment of first determined pathological condition(s) diagnosed in a mammalian host by eliciting a T cell immune response against a first group of epitopes contained in said polypeptide(s) and in the prophylaxis against second determined pathological condition(s) in the same mammalian host by eliciting a T cell memory immune response against a second group of epitopes contained in said polypeptide(s), said immune responses being obtained after administration of said vector-carried polypeptide(s) into said host, wherein said prophylaxis against second determined pathological condition(s) is not observed when said second group of epitopes is not contained in said administered vector-carried polypeptide(s). 1. Vector-carried polypeptide(s) , wherein said vector carrying the polypeptide(s) consists in a CyaA protein or a fragment thereof suitable to present said polypeptide(s) to the immune system in a mammalian host , for use (i) in the immunotherapeutic treatment of first determined pathological condition(s) diagnosed in said mammalian host by eliciting a T cell immune response against a first group of epitopes contained in said polypeptide(s) and (ii) in the prophylaxis against second determined pathological condition(s) in the same mammalian host by eliciting a T cell memory immune response against a second group of epitopes contained in said polypeptide(s) , said immune responses being obtained after administration of said vector-carried polypeptide(s) into said host , wherein said prophylaxis against second determined pathological condition(s) is not observed when said second group of epitopes is not contained in said administered vector-carried polypeptide(s).2. The vector-carried polypeptide according to claim 1 , for further use (iii) in the prevention against the re-occurrence of said first determined pathological condition(s) claim 1 , by eliciting a T ...

Activity of Fe-S Cluster Requiring Proteins

The present invention is related to a recombinant host cell, in particular a yeast cell, comprising a dihydroxy-acid dehydratase polypeptide. The invention is also related to a recombinant host cell having increased specific activity of the dihydroxy-acid dehydratase polypeptide as a result of increased expression of the polypeptide, modulation of the Fe—S cluster biosynthesis of the cell, or a combination thereof. The present invention also includes methods of using the host cells, as well as, methods for identifying polypeptides that increase the flux in an Fe—S cluster biosynthesis pathway in a host cell. 131-. (canceled)32. A method of increasing the activity of an Fe—S cluster requiring protein in a recombinant host cell , comprising:(a) providing a recombinant host cell comprising an Fe—S cluster requiring protein;(b) changing the expression or activity of a polypeptide affecting Fe—S cluster biosynthesis in the host cell; and(c) growing the host cell of (b) under conditions whereby the activity of the Fe—S cluster requiring protein is increased.3362-. (canceled)63. The method of claim 32 , wherein the increase in activity is an amount greater than 10%.64. The method of claim 32 , wherein the increase in activity is greater than 5-fold.65. The method of claim 32 , wherein the Fe—S cluster requiring protein has dihydroxy-acid dehydratase activity (DHAD).66. The method of claim 32 , wherein the recombinant host cell comprises at least one heterologous polynucleotide encoding a polypeptide having DHAD activity.67. The method of claim 66 , wherein the polypeptide having DHAD activity is expressed in the cytosol of the host cell.68. The method of claim 66 , wherein the at least one heterologous polynucleotide encoding a polypeptide having DHAD activity is expressed in multiple copies.69. The recombinant host cell of claim 66 , wherein the at least one heterologous polynucleotide encoding a polypeptide having DHAD activity is integrated at least once in the ...

Method for producing terpenes

The present invention relates to a method for producing terpenes in fungi, wherein a terpene biosynthetic gene cluster having terpene biosynthetic genes and regulatory regions operably linked to said genes is activated. The invention relates also to a terpene biosynthetic gene cluster and regulatory regions of such terpene biosynthetic gene cluster usable is production of terpenes, use of regulator for regulating the terpene production and use of FGSC A4 for producing terpenes. The method of invention provides higher yields of enriched terpene product without essential amount of side-products. 1. Method for producing terpenes in fungi comprising the steps of:(a) providing a transcription factor activating a terpene biosynthetic gene cluster having terpene biosynthetic genes and regulatory regions operably linked to said genes, the transcription factor having a sequence SEQ ID NO:74, or a sequence showing at least 80% identity to SEQ ID NO: 74;(b) operably linking said transcription factor to a promoter;(c) transforming the transcription factor of item (a) operably linked to the promoter of item (b) to a host cell carrying a terpene biosynthetic gene cluster as described in item (a);(d) cultivating said host in conditions allowing the expression of the transcription factor activating the cluster; and optionally(e) recovering the terpene product.2. The method of claim 1 , wherein terpene is terpenoid.3. The method of claim 1 , wherein terpene is ent-pimara-8(14) claim 1 ,15-diene or its derivative.4. The method of claim 1 , wherein the host cell of item (c) carries the terpene biosynthetic gene cluster having terpene metabolite synthase genes of item (a).5. The method of claim 1 , wherein the terpene biosynthetic gene cluster having terpene metabolite synthase genes is transformed to a host cell.6. The method of claim 1 , wherein a gene encoding a transporter protein is included to the production host.7. (canceled)8. The method of claim 1 , wherein the transcription ...

Aldolases, Nucleic Acids Encoding Them and Methods for Making and Using Them

This invention relates to polypeptides having aldolase activity, including pyruvate activity such as, without limitation, HMG and/or KHG aldolase activity, polynucleotides encoding these polypeptides, and methods of making and using these polynucleotides and polypeptides. In some embodiments, the invention is directed to polypeptides having aldolase activity, including pyruvate activity such as, without limitation, HMG and/or KHG aldolase activity, including thermostable and thermotolerant activity, and polynucleotides encoding these enzymes, and making and using these polynucleotides and polypeptides. The polypeptides in accordance with the invention can be used in a variety of pharmaceutical, agricultural and industrial contexts. In some embodiments, the invention provides polypeptides and biosynthetic pathways that are useful in the production of R-2-hydroxy 2-(indol-3ylmethyl)-4-keto glutaric acid (R-MP) and certain stereoisomers of monatin, such as R,R and S,R monatin, and salts thereof, as well as certain stereoisomers of monatin derivatives, such as the R,R and S,R configurations, and salts thereof. 1. An isolated , synthetic , or recombinant nucleic acid that encodes a polypeptide having aldolase activity , wherein the nucleic acid is selected from the group consisting of:(a) a variant nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more or complete (100%) sequence identity to the full length of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:71, SEQ ID ...

Production of Volatile Dienes by Enzymatic Dehydration of Light Alkenols

Described is a method for generating conjugated dienes through a biological process. More specifically, the application describes a method for producing conjugated dienes (for example butadiene, isoprene or dimethylbutadiene) from light alkenols via enzymatic dehydration, in particular by making use of an alkenol dehydratase.

SYSTEMS USING CELL CULTURE FOR PRODUCTION OF ISOPRENE

The invention features methods for producing isoprene from cultured cells. The invention also provides compositions that include these cultured cells. The invention provides isoprene compositions, such as compositions with increased amount of isoprene or increased purity. Additionally, the invention provides methods of producing isoprene by culturing cells under conditions suitable for isoprene production while maintaining cell viability and/or metabolic activity. 1. A method of producing isoprene , the method comprising:a) culturing cells comprising one or more nucleic acids encoding for isoprene synthase polypeptide under suitable conditions for production of isoprene; and{'sub': 'wcm', 'sup': '−18', 'b) producing isoprene, wherein the cells produce greater than about 400 nmole/g/hour of isoprene, and wherein the carbon dioxide evolution rate of the cells is greater than about 1×10mmol/L/hour.'}2. The method of claim 1 , further comprising recovering the isoprene.3. The method of claim 1 , wherein the cells further comprise one or more heterologous nucleic acids or one or more additional copies of an endogenous nucleic acid encoding an isoprene synthase polypeptide.4. The method of claim 1 , wherein the cells further comprise one or more heterologous nucleic acids or one or more additional copies of an endogenous nucleic acid encoding an IDI polypeptide claim 1 , an MVA pathway enzyme claim 1 , or a DXP pathway enzyme.5. The method of wherein the MVA pathway enzyme is mevalonate kinase.6PuerariaPopulusPopulus alba×Populus tremula.. The method of claim 1 , wherein the isoprene synthase polypeptide is from or or a hybrid claim 1 ,7Pueraria montanaPueraria lobata, Populus tremuloides, Populus alba, Populus nigraPopulus trichocarpa.. The method of claim 6 , wherein the isoprene synthase polypeptide is selected from the group consisting of or claim 6 , and8StreptomycesEscherichiaPantoeaTrichodermaAspergillus. The method of claim 1 , wherein the cells are gram-positive ...

VALENCENE SYNTHASE

The present invention relates to a valencene synthase, to a nucleic acid encoding such valencene synthase, to a host cell comprising said encoding nucleic acid sequence and to a method for preparing valencene, comprising converting farnesyl diphosphate to valencene in the presence of a valencene synthase according to the invention. 1. A valencene synthase comprising an amino acid sequence as shown in SEQ ID NO: 2 , SEQ ID NO: 4 , or a functional homologue of any of these sequences , said homologue being a valencene synthase comprising an amino acid sequence which has a sequence identity of at least 40% with SEQ ID NO: 2 or SEQ ID NO: 4.2. The valencene synthase according to claim 1 , having at least 60% claim 1 , at least 70% claim 1 , at least 80% claim 1 , at least 90% claim 1 , at least 95% claim 1 , or at least 98% sequence identity with SEQ ID NO: 2 or SEQ ID NO: 4.3. A nucleic acid claim 1 , comprising a nucleic acid sequence encoding a valencene synthase according to claim 1 , a complementary sequence thereof claim 1 , or comprising a nucleic acid sequence hybridising with a nucleic acid sequence encoding a valencene synthase according to under stringent conditions.4. A nucleic acid according to claim 3 , wherein the nucleic acid comprises a nucleic acid sequence as shown in SEQ ID NO: 1 claim 3 , SEQ ID NO: 3 claim 3 , SEQ ID NO: 18 claim 3 , SEQ ID NO: 19 or another nucleic acid sequence encoding a valencene synthase comprising a nucleic acid sequence having a sequence identity of at least 40% claim 3 , at least 60% claim 3 , at least 70% claim 3 , at least 80% claim 3 , at least 90% or at least 95% with any of the sequences shown in SEQ ID NO: 1 claim 3 , SEQ ID NO: 3 claim 3 , SEQ ID NO: 18 claim 3 , or SEQ ID NO: 19.5. An expression vector comprising a nucleic acid according to .6. A host cell claim 5 , which may be an organism per se or part of a multi-cellular organism claim 5 , said host cell comprising an expression vector according to claim 5 , ...

Polymerization of isoprene from renewable resources

This invention relates to compositions and methods for producing polymers of isoprene derived from renewable resources, such as isoprene produced from cultured cells that use renewable carbon sources. A starting isoprene composition, such as a bioisoprene composition, is distinguished from petroleum based isoprene by its purity profile (such as lower levels of certain C 5 hydrocarbons other than isoprene, presence of certain compounds associated with the biological process for production) and the relative content of the carbon isotopes. Polymers obtained by polymerization of such starting isoprene composition according to this invention, such as a polyisoprene homopolymer or a copolymer having repeat units that are derived from isoprene, are distinguishable from isoprene containing polymers from petrochemical resources.

Fungi and Products Thereof

The present invention provides substantially purified or isolated fungi of spp. or spp., plants infected with said fungi, organic compounds produced by said fungi, and related nucleic acids, polypeptides and methods. 117-. (canceled)18NodulisporiumAscocoryne. A substantially purified or isolated fungus , wherein said fungus consists of (1) a spp. that produces at least one volatile terpenoid compound when grown in potato dextrose culture medium or (2) an spp , that produces at least organic compound that is liquid at room temperature when grown in potato dextrose culture medium.19. The fungus according to claim 18 , wherein said fungus includes a nucleic acid encoding a terpene synthase claim 18 , said nucleic acid including a nucleic acid sequence selected from the group consisting of sequences shown in claim 18 , hereto claim 18 , and functionally active fragments and variants thereof.20NodulisporiumAscocoryne. The fungus according to claim 18 , wherein said fungus is selected from the group consisting of spp. or spp. selected from the group consisting of Dandenong Ranges isolate 1 and Yarra Ranges isolates 7 claim 18 , 10 claim 18 , 11 claim 18 , 12 claim 18 , 13 and 15 and Otway Ranges isolates 1 claim 18 , 3 claim 18 , 4 and 5.21. A plant inoculated with the fungus according to claim 20 , said plant comprising a fungus-free host plant stably infected with said fungus.22. A plant inoculated with the fungus according to claim 18 , said plant comprising a fungus-free host plant stably infected with said fungus.23. A method of producing an organic compound claim 18 , said method including growing a fungus according to in a culture medium under conditions suitable to produce said organic compound claim 18 , and recovering the organic compound produced by the fungus.24. The method according to claim 23 , wherein said culture medium includes a source of carbohydrates claim 23 , and wherein said fungus is grown under aerobic or anaerobic conditions.25. The method ...

COMPOSITIONS AND METHODS FOR PRODUCTION OF MYRCENE

Provided herein are compositions and methods for producing myrcene by culturing genetically modified microbial host cells that express a myrcene synthase and optionally a geranyl pyroplosphate synthase. Also provided herein are isolated nucleic acid molecules that encode myrcene synthase variants derived from the species myrcene synthase, which comprise one or more amino acid substitutions that improve in vivo performance of myrcene synthase in genetically modified microbial host cells. Also provided herein are isolated myrcene synthase variants that exhibit an improved activity for converting geranyl diphosphate into myrcene. 137.-. (canceled)38. A genetically modified microbial host cell comprising:{'i': 'Ocimum', 'claim-text': (i) an amino acid sequence of SEQ ID NO: 2; or', '(ii) an amino acid sequence that has at least 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%), at least about 97%, at least about 8%, at least about 99% sequence identity to SEQ ID NO: 2; and, '(a) a heterologous nucleic acid molecule encoding an species myrcene synthase that comprises(b) a heterologous nucleic acid molecule encoding a geranyl pyrophosphate synthase.39. The genetically modified microbial host cell of claim 38 , wherein the heterologous nucleic acid encodes a geranyl pyrophosphate synthase derived from a bacterium.40Streptomyces aculeolatus. The genetically modified host cell of claim 39 , wherein the geranyl pyrophosphate synthase is derived from a geranyl pyrophosphate synthase.41Streptomyces aculeolatus. The genetically modified host cell of claim 40 , wherein the geranyl pyrophosphate synthase comprises an amino acid sequence having SEQ ID NO: 7 or a variant thereof that has at least about 70% claim 40 , at least about 75% claim 40 , at least about 80% claim 40 , at least about 85% claim 40 , at least about 90% claim 40 , at least about 95% claim 40 , at least about 96% claim 40 , at least about ...

Production of isoprene, isoprenoid, and isoprenoid precursors using an alternative lower mevalonate pathway

The invention provides for compositions and methods for the production of isoprene, isoprenoid precursor, and/or isoprenoids in cells via the expression (e.g., heterologous expression) of phosphomevalonate decarboxylases and/or isopentenyl kinases.

METHODS OF PRODUCING PROTOPORPHYRIN IX AND BACTERIAL MUTANTS THEREFOR

The presently disclosed inventive concepts are directed in certain embodiments to a method of producing protoporphyrin IX by (1) cultivating a strain of bacteria in a culture medium under conditions suitable for growth thereof, and (2) recovering the protoporphyrin IX from the culture medium. The strain of bacteria comprises at least one mutant hemH gene which is incapable of normal expression, thereby causing an accumulation of protoporphyrin IX. In certain embodiments of the method, the strain of bacteria is a strain of , and more specifically may be PV-4. In certain embodiments, the mutant hemH gene of the strain of bacteria may be a mutant of shew_2229 and/or of shew_1140. In other embodiments, the presently disclosed inventive concepts are directed to mutant strains of bacteria having at least one mutant hemH gene which is incapable of normal expression, thereby causing an accumulation of protoporphyrin IX during cultivation of the bacteria. In certain embodiments the strain of bacteria is a strain of , and more specifically may be PV-4. In certain embodiments, the mutant hemH gene of the strain of bacteria may be a mutant of shew_2229 and/or shew_1140. 1. A method of producing protoporphyrin IX , comprising:{'i': Shewanella', 'Shewanella, 'cultivating a strain of bacteria in a culture medium under conditions suitable for growth thereof, the strain of bacteria comprising at least one mutant hemH gene which is incapable of normal expression, thereby causing an accumulation of protoporphyrin IX; and'}recovering the protoporphyrin IX from the culture medium.2ShewanellaS. loihica.. The method of claim 1 , wherein the strain of bacteria is a strain of3ShewanellaS. loihica. The method of claim 1 , wherein the strain of bacteria is PV-4.4. The method of claim 1 , wherein the mutant hemH gene is a mutant of shew2229 and/or a mutant of shew1140.5. The method of claim 1 , wherein the protoporphyrin IX produced in the culture medium is recoverable in an amount of at least ...

METABOLIC ENGINEERING FOR MICROBIAL PRODUCTION OF TERPENOID PRODUCTS

The invention relates to methods and bacterial strains for making terpene and terpenoid products, the bacterial strains having improved carbon pull through the MEP pathway and to a downstream recombinant synthesis pathway. 1. A method for production of a terpene or terpenoid product , comprising:providing a bacterial strain that produces isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) through an upstream methylerythritol phosphate pathway (MEP pathway) and converts the IPP and DMAPP to a terpene or terpenoid product through a downstream synthesis pathway;wherein IspG and IspH are overexpressed in the bacterial strain such that IspG activity and IspH activity are balanced to provide increased carbon flux to 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate (HMBPP) intermediate, while preventing accumulation of HMBPP at an amount that feeds back and reduces MEP pathway flux and terpene or terpenoid productivity, the strain optionally comprising one or more genetic modifications that enhance supply and/or transfer of electrons through the MEP pathway and/or to terpene and terpenoid products, and culturing the bacterial strain to produce the terpene or terpenoid product.2EscherichiaBacillusCorynebacteriumRhodobacterZymomonasVibrioPseudomonas. The method of claim 1 , wherein the bacterial strain is a bacteria selected from spp. claim 1 , spp. claim 1 , spp. claim 1 , spp. claim 1 , spp. claim 1 , spp. claim 1 , and spp.3Escherichia coli, Bacillus subtilis, Corynebacterium glutamicum, Rhodobacter capsulatus, Rhodobacter sphaeroides, Zvmomonas mobilis, Vibrio natriegensPseudomonas putida.. The method of claim 2 , wherein the bacterial strain is a species selected from claim 2 , or4E. coli.. The method of claim 3 , wherein the bacterial strain is5. The method of any one of to claim 3 , wherein the bacterial strain expresses dxs claim 3 , ispD claim 3 , ispF claim 3 , and idi as recombinant genes claim 3 , which are optionally expressed as an operon.6. The ...

Metabolic engineering for microbial production of terpenoid products

The invention relates to methods and bacterial strains for making terpene and terpenoid products, the bacterial strains having improved carbon pull through the MEP pathway and to a downstream recombinant synthesis pathway.

METHODS AND COMPOSITIONS FOR PRODUCTION OF AROMATIC AND OTHER COMPOUNDS IN YEAST

The present disclosure includes methods and components for production of valuable industrial compounds in yeast. In an embodiment, the present invention provides a nucleic acid construct with increased stability for gene expression or gene editing comprising: a nucleic acid sequence encoding one or more of SEQ ID NO: 1-8 (CENs 1-8); and one or more regulatory elements functional in a yeast cell. In an embodiment of the present invention the nucleic acid constructs are vectors, preferably episomal vectors. High expression promoters, as well as methods for increasing production of compounds such as aromatics are disclosed. 1. A nucleic acid construct for gene expression or gene editing in yeast comprising:a nucleic acid sequence encoding one or more of SEQ ID NO: 1-8 (CENs 1-8); andone or more regulatory elements functional in a yeast cell.2. The nucleic acid construct of claim 1 , wherein said nucleic acid construct further comprises an autonomously replicating sequence (ARS).3. The nucleic acid construct of claim 1 , wherein said nucleic acid construct includes a multiple cloning site for insertion of gene of interest operably linked to the regulatory elements.4. The nucleic acid construct of claim 1 , wherein the regulatory element is a promoter.5. The nucleic acid construct of claim 1 , wherein said regulatory element is a terminator sequence.6. The nucleic acid construct of wherein the gene of interest is inserted in a multiple cloning site.7. A vector comprising the nucleic acid construct of .8. The vector of claim 7 , wherein said vector is a plasmid vector.9. The vector of claim 7 , wherein said vector is an episomal vector.10. The vector of claim 8 , wherein said plasmid vector has increased stability compared to the same plasmid vector lacking one or more of SEQ ID NO:1-8 (CENs 1-8).11. A cell claim 1 , tissue claim 1 , or organ comprising the nucleic acid construct of .12. The nucleic acid construct of claim 4 , wherein the promoters are selected from the ...

Production of manool

Provided herein are methods of producing (+)-manool comprising: contacting geranylgeranyl diphosphate with an copalyl diphosphate (CPP) synthase to form a (9S, 10S)-copalyl diphosphate wherein the CPP synthase comprises an amino acid sequence having at least 90%, 95%, 98%, 99% and 100% sequence identity to a polypeptide selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO:2; and contacting the CPP with a sclareol synthase enzyme to form (+)-manool.

METHODS, MATERIALS, SYNTHETIC HOSTS AND REAGENTS FOR THE BIOSYNTHESIS OF HYDROCARBONS AND DERIVATIVES THEREOF

Genetically engineered hosts and methods for their production and use in synthesizing hydrocarbons are provided. 1: A genetically engineered host capable of producing hydrocarbons via a mevalonate (MVA) pathway.3: The genetically engineered host of comprising at least one genome-integrated synthetic operon encoding an enzyme of the MVA pathway.4: The genetically engineered host of comprising a genome-integrated synthetic operon encoding a plurality of enzymes of the MVA pathway.5: The genetically engineered host of encoding one or more enzymes selected from acetoacetyl-CoA C-acetyltransferase (AACT) claim 1 , HMG-CoA reductase (HMGR) claim 1 , hydroxymethylglutaryl-CoA synthase (HMGS) claim 1 , mevalonate kinase (MVK) claim 1 , phosphomevalonate kinase (MPK0 claim 1 , mevalonate diphosphate decarboxylase (MDD) claim 1 , isopentenyl diphosphate isomerase (IDI) and isoprene synthase (ISPS).6: The genetically engineered host of comprising a genome-integrated synthetic operon encoding one or more of enzymes of the upper MVA pathway.7Enterococcus faecalis: The genetically engineered host of comprising a genome-integrated synthetic operon encoding one or more of the enzymes of the upper MVA pathway.8: The genetically engineered host of comprising an exogenous nucleic acid sequence encoding a polypeptide having AACT claim 6 , HMGR or HGMS enzyme activity.9: The genetically engineered host of wherein the polypeptide having AACT claim 8 , HMGR or HGMS enzyme activity:(i) has at least 70% sequence identity to an amino acid sequence set forth in SEQ ID NO: 1, 56, 58, 60, 62, 64 66, 2, 68, 70, 72 or 74 or a functional fragment thereof;(ii) comprises an amino acid sequence set forth in SEQ ID NO: 1, 56, 58, 60, 62, 64 66, 2, 68, 70, 72 or 74 or a functional fragment thereof;(iii) has at least 70% sequence identity to the nucleic acid sequence set forth in SEQ ID NO: 23, 55, 57, 59, 61, 63, 65, 24, 67, 69, 71 or 73 or a functional fragment thereof; or(iv) comprises SEQ ID NO: 23, ...

METHODS, SYNTHETIC HOSTS AND REAGENTS FOR THE BIOSYNTHESIS OF HYDROCARBONS

Systems, networks, methods, compositions and recombinant hosts for biosynthesizing hydrocarbons from a feedstock, such as gas, are provided. 1: A method for biosynthesising a hydrocarbon in a recombinant host , said method comprising:providing a fermentation reactor, the fermentation reactor comprising at least one recombinant host, wherein said recombinant host comprises an exogenous nucleic acid sequence encoding a polypeptide having an enzyme activity of EC 2.2.1.7, andproviding a stream comprising a gas or a biological or nonbiological feedstock to the fermentation reactor, andoperating the fermentation reactor at conditions for said recombinant host to metabolize the gas or feedstock and produce the hydrocarbon.3: The method according to claim 1 , wherein said polypeptide having an enzyme activity of EC 2.2.1.7 converts glyceraldehyde-3-phosphate and pyruvate to 1 deoxy-d-xylulose-phosphate.4: The method according to claim 1 , wherein said polypeptide having an enzyme activity of EC 2.2.1.7 is:(i) encoded by a nucleic acid sequence having at least 49% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:4 or 5 or a functional fragment thereof;(ii) encoded by a nucleic acid sequence comprising the nucleic acid sequence set forth in SEQ ID NO:4 or 5 or a functional fragment thereof;(iii) has at least 49% sequence identity to the amino acid sequence set forth in SEQ ID NO:1 or 2 or a functional fragment thereof; or(iv) comprises the amino acid sequence set forth in SEQ ID NO: 1 or 2 or a functional fragment thereof.57-. (canceled)8: The method according to claim 1 , wherein the recombinant host further comprises an exogenous nucleic acid sequence encoding a polypeptide having an enzyme activity of EC 4.2.3.27.10: The method according to claim 8 , wherein said polypeptide having an enzyme activity of EC 4.2.3.27 is:(i) encoded by a nucleic acid sequence having at least 70% sequence identity to the nucleic acid sequence set forth in SEQ ID NO: 6 or ...

PRODUCTION OF ISOPRENE UNDER REDUCED OXYGEN INLET LEVELS

This invention relates to methods for producing isoprene by culturing recombinant cells (e.g., cells engineered to produce isoprene) under reduced oxygen inlet levels. 132-. (canceled)33. A method for producing isoprene comprising (a) culturing a recombinant fungal host cell comprising a heterologous nucleic acid encoding isoprene synthase under reduced oxygen inlet levels , wherein the reduced oxygen inlet levels comprise between about 4% to about 15% oxygen and wherein the cell is in fermentation or production phase; and (b) producing isoprene.34. The method of claim 33 , further comprising recovering the isoprene.35. The method of claim 33 , wherein the reduced oxygen inlet level is between about 5% to about 15% oxygen.36. The method of claim 35 , wherein the reduced oxygen inlet level is between about 7% to about 10% oxygen.37. The method of claim 36 , wherein the reduced oxygen inlet level is about 7.7% oxygen.38. The method of claim 36 , wherein the reduced oxygen inlet level is about 9.3% oxygen.39. The method of claim 33 , wherein the isoprene synthase is a plant isoprene synthase.40eucalyptus. The method of claim 39 , wherein the plant isoprene synthase is a poplar isoprene synthase claim 39 , a kudzu isoprene synthase claim 39 , a willow isoprene synthase claim 39 , or a isoprene synthase.41PuerariaPopulusPopulus alba×Populus tremula.. The method of claim 39 , wherein the plant isoprene synthase is an isoprene synthase from or or a hybrid claim 39 ,42Pueraria montana, Pueraria lobata, Populus tremuloides, Populus alba, Populus nigraPopulus trichocarpa.. The method of claim 39 , wherein the plant isoprene synthase polypeptide is selected from the group consisting of claim 39 , and43. The method of claim 33 , wherein the isoprene synthase is an isoprene synthase variant.44. The method of claim 33 , wherein the cell further comprises a heterologous nucleic acid encoding for one or more mevalonate (MVA) pathway polypeptides and/or one or more 1-deoxy-d- ...

Valencene Synthase Polypeptides, Encoding Nucleic Acid Molecules And Uses Thereof

Provided are valencene synthase polypeptides, nucleic acid molecules encoding the valencene synthases, host cells containing the nucleic acids and methods for producing products whose production is catalyzed by the polypeptides. Also provided are methods for producing valencene and nootkatone.

USE OF GENETICALLY ENGINEERED STRAIN VNP20009-M IN PREVENTING AND TREATING CANCER METASTASIS

The present disclosure provides uses of genetically engineered attenuated strain VNP20009-M in preventing and treating cancer metastasis. The genetically engineered strain VNP20009-M is targeted to cancer cells and has a significant effect of inhibiting metastasis and growth. VNP20009-M can be used to prepare medicaments for the prevention and treatment of tumor metastasis. 1Salmonella typhimurium. A method for preventing or treating metastasis of a cancer in a subject , the method comprising administering a therapeutically effective amount of a genetically engineered strain VNP20009-M to the subject , wherein the genetically engineered strain VNP20009-M is an attenuated VNP20009 comprising an L-methioninase gene.2Salmonella typhimurium. The method according to claim 1 , wherein the genetically engineered bacterium VNP20009-M is an attenuated VNP20009 which comprises a vector comprising the L-methioninase gene.3. The method according to claim 1 , wherein the cancer is selected from the group consisting of lung cancer claim 1 , breast cancer claim 1 , prostate cancer claim 1 , pancreatic cancer claim 1 , liver cancer claim 1 , colon cancer claim 1 , rectal cancer claim 1 , gastric cancer claim 1 , esophageal cancer claim 1 , laryngeal cancer claim 1 , leukemia claim 1 , lymphoma claim 1 , melanoma claim 1 , uterine cancer claim 1 , ovarian cancer claim 1 , skin cancer claim 1 , bronchial carcinoma claim 1 , bronchiolar carcinoma claim 1 , urethral cancer claim 1 , kidney cancer claim 1 , oral cancer claim 1 , vaginal cancer claim 1 , cholangiocarcinoma claim 1 , bladder cancer and nasopharyngeal carcinoma.4Salmonella typhimurium. The method according to claim 2 , wherein the genetically engineered strain VNP20009-M is constructed according to the following method: inserting the L-methioninase gene into the plasmid to obtain an L-methioninase expression plasmid; electro-transforming the L-methioninase expression plasmid to attenuated VNP20009; and obtaining the ...

COMPOSITIONS AND METHODS OF USING CHONDROITINASE ABCI MUTANTS

One aspect of the present invention relates to mutants of chondroitinase ABCI. Such chondroitinase ABCI mutants exhibit altered chondroitin lyase activity or increased resistance to inactivation from stressors including exposure to UV light or heat. Methods of using chondroitinase ABCI mutant enzymes are also provided. 1. An isolated nucleic acid comprising a cDNA sequence that encodes for a mutant chondroitinase ABC I polypeptide selected from the group consisting of SEQ ID NO: 1 , SEQ ID NO: 2 , SEQ ID NO: 3 , SEQ ID NO: 4 , SEQ ID NO: 5 , SEQ ID NO: 6 , and SEQ ID NO: 7.2. The nucleic acid of claim 1 , wherein the cDNA sequence encodes for the mutant chondroitinase ABC I polypeptide of SEQ ID NO:1.3. The nucleic acid of claim 1 , wherein the cDNA sequence encodes for the mutant chondroitinase ABC I polypeptide of SEQ ID NO:2.4. The nucleic acid of claim 1 , wherein the cDNA sequence encodes for the mutant chondroitinase ABC I polypeptide of SEQ ID NO:3.5. The nucleic acid of claim 1 , wherein the cDNA sequence encodes for the mutant chondroitinase ABC I polypeptide of SEQ ID NO:4.6. The nucleic acid of claim 1 , wherein the cDNA sequence encodes for the mutant chondroitinase ABC I polypeptide of SEQ ID NO:5.7. The nucleic acid of claim 1 , wherein the cDNA sequence encodes for the mutant chondroitinase ABC I polypeptide of SEQ ID NO:6.8. The nucleic acid of claim 1 , wherein the cDNA sequence encodes for the mutant chondroitinase ABC I polypeptide of SEQ ID NO:7.9. A composition comprising an isolated nucleic acid sequence that encodes for a mutant chondroitinase ABC I polypeptide selected from the group consisting of SEQ ID NO: 1 claim 1 , SEQ ID NO: 2 claim 1 , SEQ ID NO: 3 claim 1 , SEQ ID NO: 4 claim 1 , SEQ ID NO: 5 claim 1 , SEQ ID NO: 6 claim 1 , and SEQ ID NO: 7.10. The composition of claim 9 , wherein the isolated nucleic acid sequence is present in effective amount. This application is a continuation application of U.S. application Ser. No. 14/480,486, ...

TRANSFORMANT FOR EXPRESSING CIS-PRENYLTRANFERASE AND NOGO B RECEPTOR

Provided are a transformant produced by introducing a gene coding for a cis-prenyltransferase and a gene coding for a Nogo-B receptor, which are considered to be involved in polyisoprenoid biosynthesis, into a host to allow the host to express the cis-prenyltransferase and the Nogo-B receptor, and a method for producing a polyisoprenoid using the transformant. The present invention relates to a transformant produced by introducing a gene coding for a cis-prenyltransferase and a gene coding for a Nogo-B receptor into a host to allow the host to express the cis-prenyltransferase and the Nogo-B receptor. 1. A transformant , produced by introducing a gene coding for a cis-prenyltransferase and a gene coding for a Nogo-B receptor into a host to allow the host to express the cis-prenyltransferase and the Nogo-B receptor.2. The transformant according to claim 1 ,wherein at least one of the gene coding for a cis-prenyltransferase or the gene coding for a Nogo-B receptor is derived from a plant.3. The transformant according to claim 2 ,wherein at least one of the gene coding for a cis-prenyltransferase or the gene coding for a Nogo-B receptor is derived from an isoprenoid-producing plant.4. The transformant according to claim 3 ,{'i': 'Hevea brasiliensis.', 'wherein at least one of the gene coding for a cis-prenyltransferase or the gene coding for a Nogo-B receptor is derived from'}5. The transformant according to claim 1 ,wherein the gene coding for a Nogo-B receptor is either of the following DNAs:[3] a DNA comprising the nucleotide sequence of SEQ ID NO:5; and[4] a DNA capable of hybridizing to a DNA comprising a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO:5 under stringent conditions.6. The transformant according to claim 1 ,wherein the gene coding for a cis-prenyltransferase is either of the following DNAs:[1] a DNA comprising the nucleotide sequence of SEQ ID NO:1 or 3; and[2] a DNA capable of hybridizing to a DNA comprising a nucleotide ...

Methods for Isoprene and Pinene Production in Cyanobacteria

Methods of isoprenoid production are provided by the present invention. In particular, transgenic sp. PCC 7002 cyanobacteria and methods for producing isoprene and pinene using a host transgenic sp. PCC 7002 cyanobacterium are provided. 1. A method for pinene production , the method comprising the steps of:{'i': 'Synechococcus', '(a) obtaining a host transgenic sp. PCC 7002 cyanobacterium comprising one or more transgenes encoding geranyl diphosphate synthase (GPPS), and mono-terpene synthase (mono-TPS); and'}{'sup': −1', '−1, '(b) observing pinene production by the cyanobacterium, wherein pinene is produced at a rate of at least about 330 μg gDWh.'}2. The method of claim 1 , wherein pinene is produced at a rate of at least about 660 μg gDWh.3. The method of claim 1 , wherein pinene is produced at a rate of at least about 2000 μg gDWh.4. The method of claim 1 , wherein pinene is produced at a rate of at least about 4000 μg gDWh.5Synechocystis. The method of claim 1 , wherein the cyanobacterium further comprises a promoter derived from sp. PCC 6803.6Synechocystis. The method of claim 5 , wherein the promoter derived from sp. PCC 6803 is PcpcB.7Synechocystis. The method of claim 5 , wherein the promoter derived from sp. PCC 6803 is a synthetic PpsaA/B promoter.8. The method of claim 1 , wherein the one or more transgenes comprises codons preferred for expression in the cyanobacterium.9Artemisia. The method of claim 1 , wherein the one or more transgenes encodes a protein identical to that isolated from an species.10ArtemisiaArtemisia annua.. The method of claim 9 , wherein the protein isolated from an species is mono-TPS from11. The method of claim 1 , wherein the cyanobacterium of step (a) further comprises at least one transgene selected from the group consisting of a transgene encoding hydroxymethylbutenyl diphosphate reductase (HDR) and 1-deoxy-D-xylulose-5-phosphate synthase (DXS).12. The method of claim 1 , wherein pinene is produced under high COconditions.13. ...

Herbicide-resistant rice plants, polynucleotides encoding herbicide-resistant acetohydroxyacid synthase large subunit proteins, and methods of use

Herbicide-resistant rice plants, isolated polynucleotides that encode herbicide resistant and wild-type acetohydroxy-acid synthase large subunit 1 (AHASL1) polypeptides, and the amino acid sequences of these polypeptides, are described. Expression cassettes and transformation vectors comprising the polynucleotides of the invention, as well as plants and host cells transformed with the polynucleotides, are described. Methods of using the polynucleotides to enhance the resistance of plants to imidazolinone herbicides, and methods for controlling weeds in the vicinity of herbicide-resistant plants are also described.

Biosynthesis of Forskolin and Related Compounds

The invention described materials and methods for producing oxidised 13R-MO, such as forskolin. In particular, the invention describes P450s involved in oxidation of 13R-MO including CYP76AH8, CYP76AH11, CYP76AH15, CYP76AH17, CYP71D381 and CYP76AH9. Host organisms expressing one or more of these P450s are useful in the production of oxidized 13R-MO. 1. A host organism , comprising a heterologous nucleic acid encoding an enzyme capable of catalysing hydroxylation of 13R-manoyl oxide (13R-MO) and/or an oxidised 13R-MO derivative at the 11-position , wherein the oxidised 13R-MO carries a —H at the 11-position; and/or catalysing oxidation of a hydroxyl group to form an oxo-group at the 11-position of 11-hydroxyl-13R-MO and/or an oxidised 11-hydroxyl-13R-MO , wherein the enzyme comprises:(a) CYP76AH8 having at least 70% identity to the amino acid sequence set forth in SEQ ID NO:1 or a functional homologue thereof;(b) CYP76AH17 having at least 70% identity to the amino acid sequence set forth in SEQ ID NO:10 or a functional homologue thereof;(c) CYP76AH15 having at least 70% identity to the amino acid sequence set forth in SEQ ID NO:11 or a functional homologue thereof; and/or(d) CYP76AH11 having at least 70% identity to the amino acid sequence set forth in SEQ ID NO:2 or a functional homologue thereof.2. The host organism of claim 1 , further comprising:(a) a heterologous nucleic acid encoding an enzyme capable of catalysing hydroxylation of 13R-MO and/or oxidised 13R-MO at the 1-position, wherein the oxidised 13R-MO carries a —H at the 1-position;(b) a heterologous nucleic acid encoding an enzyme capable of catalysing hydroxylation of 13R-MO and/or oxidised 13R-MO at the 6-position, wherein the oxidised 13R-MO carries a —H at the 6-position;(c) a heterologous nucleic acid encoding an enzyme capable of catalysing hydroxylation of 13R-MO and/or oxidised 13R-MO at the 7-position, wherein the oxidised 13R-MO carries a —H at the 7-position; or(d) a heterologous nucleic acid ...

ENGINEERED DEOXYRIBOSE-PHOSPHATE ALDOLASES

The present invention provides engineered deoxyribose-phosphate aldolase polypeptides useful under industrial process conditions for the production of pharmaceutical and fine chemical compounds. 1. An engineered deoxyribose-phosphate aldolase comprising a polypeptide sequence having at least 85% , 86% , 87% , 88% , 89% , 90% , 91% , 92% , 93% , 94% , 95% , 96% , 97% , 98% , 99% , or more sequence identity to SEQ ID NOS: 2 , 6 , and/or 466 , or a functional fragment thereof , wherein said engineered deoxyribose-phosphate aldolase comprises at least one substitution or substitution set in said polypeptide sequence , and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 2 , 6 , and/or 466.2. The engineered deoxyribose-phosphate aldolase of claim 1 , wherein said polypeptide sequence has at least 85% claim 1 , 86% claim 1 , 87% claim 1 , 88% claim 1 , 89% claim 1 , 90% claim 1 , 91% claim 1 , 92% claim 1 , 93% claim 1 , 94% claim 1 , 95% claim 1 , 96% claim 1 , 97% claim 1 , 98% claim 1 , 99% claim 1 , or more sequence identity to SEQ ID NO:2 claim 1 , wherein said engineered deoxyribose-phosphate aldolase comprises at least one substitution or substitution set in said polypeptide sequence at one or more positions selected from 10/47/66/141/145/156 claim 1 , 2 claim 1 , 6 claim 1 , 9 claim 1 , 10/47/88/156 claim 1 , 13 claim 1 , 31 claim 1 , 46 claim 1 , 47 claim 1 , 47/134/141/212 claim 1 , 66 claim 1 , 66/88/112/134/141/143/145/212 claim 1 , 71 claim 1 , 72 claim 1 , 88 claim 1 , 94 claim 1 , 102 claim 1 , 104 claim 1 , 112 claim 1 , 116 claim 1 , 133 claim 1 , 133/173/204/235/236 claim 1 , 134 claim 1 , 145 claim 1 , 145/173 claim 1 , 147 claim 1 , 173 claim 1 , 184 claim 1 , 189 claim 1 , 197 claim 1 , 203 claim 1 , 204 claim 1 , 207 claim 1 , 226 claim 1 , 235 claim 1 , 235/236 claim 1 , and 236 claim 1 , and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: ...

USE OF N-ACETYLNEURAMINIC ACID ALDOLASE IN CATALYTIC SYNTHESIS OF N-ACETYLNEURAMINIC ACID

It discloses a use of N-acetylneuraminic acid aldolase with an amino acid sequence as shown in SEQ ID NO: 2 in catalytic synthesis of N-acetylneuraminic acid. The preparation of N-acetylneuraminic acid is to use the N-acetylneuraminic acid aldolase with the amino acid sequence as shown in SEQ ID NO: 2 as a catalyst, and N-acetylmannosamine and pyruvic acid as substrates.

Human Cystathionine Beta-Synthase Variants and Methods of Production Thereof

Human cystathionine β-synthase variants are disclosed, as well as a method to produce recombinant human cystathionine β-synthase and variants thereof. More particularly, the role of both the N-terminal and C-terminal regions of human CBS has been studied, and a variety of truncation mutants and modified CBS homologues are described. In addition, a method to express and purify recombinant human cystathionine β-synthase (CBS) and variants thereof which have only one or two additional amino acid residues at the N-terminus are described.

PLANTS HAVING ENHANCED YIELD-RELATED TRAITS AND A METHOD FOR MAKING THE SAME

The present invention relates to a method for enhancing various yield-related traits by modulating expression in a plant of a nucleic acid encoding an Ornithine Decarboxylase (ODC) polypeptide, a benzothiadiazole-induced homeodomain 1 (BIHD1) polypeptide, a MYB30, a THOM (tomato homeobox) protein, or a benzothiadiazole-induced homeodomain 2 (BIHD2) polypeptide. The present invention also concerns plants having modulated expression of such a nucleic acid, which plants have enhanced yield-related traits relative to corresponding control plants. The invention also provides constructs useful in the methods of the invention.

BIOLOGICAL DEVICES AND METHODS FOR INCREASING THE PRODUCTION OF LYCOPENE FROM PLANTS

Described herein are devices and methods for enhancing the physiological properties of plants. For example, the devices and methods described herein increase the production of lycopene, which has industrial and economic value. The lycopene produced by the devices and methods does not require the ultra purification that is common in conventional or commercial methods. The devices and methods described herein also enhance the growth rate of plants.

Pest resistant plants

The disclosure provides an isolated nucleic acid molecule encoding a 7-epizingiberene synthase, a chimeric gene comprising said nucleic acid molecule, vectors comprising the same, as well as isolated 7-epizingiberene synthase proteins themselves. In addition, transgenic plants and plant cells comprising a gene encoding a 7-epizingiberene synthase, optionally integrated in its genome, and methods for making such plants and cells, are provided. Especially Solanaceae plants and plant parts (seeds, fruit, leaves, etc.) with enhanced insect pest resistance are provided.

COMPOSITIONS AND METHODS FOR BIOLOGICAL PRODUCTION OF ISOPRENE

The present disclosure provides compositions and methods for biologically producing isoprene using methanotrophic bacteria that utilize carbon feedstock, such as methane or natural gas.

METHOD FOR COLLECTING ISOPRENOID COMPOUND CONTAINED IN FERMENTED GAS, AND METHOD FOR PRODUCING PURIFIED ISOPRENOID COMPOUND

Methods include contacting a fermented gas including isoprenoid compound with a porous adsorbent and desorbing isoprenoid compound adsorbed on the porous adsorbent. The fermented gas may be obtained by culturing a microorganism having an ability to produce isoprenoid compound.

ENGINEERED PHENYLALANINE AMMONIA LYASE POLYPEPTIDES

The present invention provides engineered phenylalanine ammonia-lyase (PAL) polypeptides and compositions thereof, as well as polynucleotides encoding the engineered phenylalanine ammonia-lyase (PAL) polypeptides. 1. An engineered polypeptide comprising an amino acid sequence having at least 90% sequence identity to reference sequence SEQ ID NO:4 , wherein said amino acid sequence comprises a glycine at position 290 , and wherein the positions in said amino acid sequence are in reference to SEQ ID NO:4.2. The engineered polypeptide of claim 1 , wherein said engineered polypeptide further comprises at least one or more of the following substitutions or substitution sets selected from 39/91/158/180/195/256/290/399/459/463 claim 1 , 39/91/158/180/290/394/399/474/522/524 claim 1 , 39/91/256/290/394/399/404/407/522/524 claim 1 , 91/158/243/256/290/399/407/459/463/474/522/524 claim 1 , and 256/290/404/407/474/522 claim 1 , wherein said positions in said amino acid sequence are in reference to SEQ ID NO:4.3. The engineered polypeptide of claim 1 , wherein said engineered polypeptide further comprises at least one or more of the following substitutions or substitution sets selected from 39/91/256/290/307/399/404/407/522/524 claim 1 , 39/91/256/290/307/404/407/524 claim 1 , 39/91/256/290/399/404/407/522 claim 1 , 39/91/290/307/407 claim 1 , 39/91/290/307/407/524 claim 1 , and 39/256/290/307/404/407 claim 1 , wherein said positions in said amino acid sequence are in reference to SEQ ID NO:4.4. An engineered polypeptide comprising an amino acid sequence having at least 90% sequence identity to reference sequence SEQ ID NO:10 claim 1 , wherein said amino acid sequence comprises a glycine at position 290 claim 1 , and wherein said engineered polypeptide further comprises at least one or more of the following substitutions or substitution sets selected from 32/54/73/305/503/521/565 claim 1 , 32/59/73/240/503/565 claim 1 , 32/59/73/240/565 claim 1 , 54/59/73/305/503/521/565 claim ...

ENGINEERED PHENYLALANINE AMMONIA LYASE POLYPEPTIDES

The present invention provides engineered phenylalanine ammonia lyase (PAL) polypeptides and compositions thereof, as well as polynucleotides encoding the engineered phenylalanine ammonia lyase (PAL) polypeptides. Methods for producing PAL enzymes are also provided. In some embodiments, the engineered PAL polypeptides are optimized to provide enhanced catalytic activities that are useful under industrial process conditions for the production of pharmaceutical compounds. 1. An engineered phenylalanine ammonia lyase comprising a polypeptide sequence having at least 85% , 86% , 87% , 88% , 89% , 90% , 91% , 92% , 93% , 94% , 95% , 96% , 97% , 98% , 99% , or more sequence identity to SEQ ID NO: 2 , 4 , 8 , 106 , 252 , 446 , 482 , 516 , 618 , 714 , 830 , 894 , and/or 988 , or a functional fragment thereof , wherein said engineered phenylalanine ammonia lyase comprises at least one substitution or substitution set in said polypeptide sequence , and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 2 , 4 , 8 , 106 , 252 , 446 , 482 , 516 , 618 , 714 , 830 , 894 , 988 , and/or 1140 , respectively.2. The engineered phenylalanine ammonia lyase of claim 1 , wherein said engineered phenylalanine ammonia lyase comprises a polypeptide sequence having at least 85% claim 1 , 86% claim 1 , 87% claim 1 , 88% claim 1 , 89% claim 1 , 90% claim 1 , 91% claim 1 , 92% claim 1 , 93% claim 1 , 94% claim 1 , 95% claim 1 , 96% claim 1 , 97% claim 1 , 98% claim 1 , 99% claim 1 , or more sequence identity to SEQ ID NO: 4 claim 1 , or a functional fragment thereof claim 1 , and wherein said engineered phenylalanine ammonia lyase comprises at least one substitution or substitution set at one or more positions selected from 80/99/104/175/220/359 claim 1 , 80/104 claim 1 , 80/104/105/172 claim 1 , 80/104/105/172/175/222/359 claim 1 , 80/104/105/172/220/222 claim 1 , 80/104/105/220 claim 1 , 80/104/105/220/222/416 claim 1 , 80/104/105/222 claim 1 ...

Tissue specific reduction of lignin

The present invention provides an expression cassette comprising a polynucleotide that encodes a protein that diverts a monolignol precursor from a lignin biosynthesis pathway in the plant, which is operably linked to a heterologous promoter. Also provided are methods of engineering a plant having reduced lignin content, as well as plant cells, plant parts, and plant tissues from such engineered plants.

Terpene Synthases for Biofuel Production and Methods Thereof

The present invention relates to terpene synthases capable of degrading precursors into biofuel compounds, such as terpenoid compounds. In one instance, a transformed organism can include such terpene synthases, as well as vectors encoding such synthases. Methods of employing such synthases and organisms are also described herein. 1. A method of treating a biomass , the method comprising: an exogenous terpenoid precursor, an exogenous enzyme configured to synthesize a terpenoid precursor, or a nucleic acid encoding the exogenous enzyme; and', 'an exogenous terpene synthase or a nucleic acid encoding the exogenous terpene synthase, wherein the exogenous terpene synthase is selected from the group consisting of a pinene synthase, a guaiene synthase, a pinene and guaiene synthase, a caryophyllene synthase, a chamigrene synthase, a chamigrene and pinene synthase, a gurjunene synthase, a gurjunene and pinene synthase, a gumunene synthase, a selinene synthase, and an isoledene synthase, or a bifunctional synthase of any of these; and, 'exposing the biomass to one or more organisms of an isolated, genetically engineered organism, wherein the organism comprisesisolating one or more terpenoid compounds.2. The method of claim 1 , wherein the exogenous terpenoid precursor is selected from the group consisting of mevalonate claim 1 , dimethylallyl pyrophosphate claim 1 , isopentenyl pyrophosphate claim 1 , farnesyl pyrophosphate claim 1 , geranyl pyrophosphate claim 1 , and geranylgeranyl pyrophosphate claim 1 , or a salt thereof.3. The method of claim 1 , wherein the organism is configured to produce one or more terpenoid compounds selected from the group consisting of a monoterpene claim 1 , a sesquiterpene claim 1 , and a diterpene.4. The method of claim 1 , wherein the nucleic acid encoding the exogenous enzyme and/or the nucleic acid encoding the exogenous terpene synthase is provided as a plasmid vector.5. The method of claim 1 , wherein the exogenous enzyme is selected ...

PRODUCTION OF SQUALENE AND/OR STEROL FROM CELL SUSPENSIONS OF FERMENTED YEAST

The present invention relates to a process for the production of squalene and/or sterol in high amounts using an alkaline solution and an organic lysis solvent at high temperature and high pressure for effectively lysing yeast cells and extracting squalene and/or sterol into an organic extraction solvent, thus obtaining squalene and/or sterolin high amount and of high purity. 1. A process for producing squalene and/or sterol comprising lysing squalene and/or sterol containing yeast cells obtained by fermentation in a culture medium under batch conditions or under continuous conditions in a lysis medium ,wherein the lysis medium comprises an aqueous suspension medium,an alkali or alkaline earth hydroxide and an organic lysis solvent,{'sup': '5', 'and wherein the lysis is performed in a closed system under elevated pressure of 2 to 50 bar (2 to 50×10Pa) and at a temperature which is above the boiling point of the lysis medium at atmospheric pressure, wherein the lysis is performed under batch conditions or under continuous conditions.'}2. The process according to claim 1 , wherein the sterol is ergosterol or lanosterol.3. The process according to any one of to claim 1 , wherein the alkali or alkaline earth hydroxide claim 1 , in solid form or in aqueous solution claim 1 , is potassium hydroxide (KOH) claim 1 , sodium hydroxide (NaOH) claim 1 , lithium hydroxide (LiOH) claim 1 , calcium hydroxide (Ca(OH)) claim 1 , magnesium hydroxide (Mg(OH)) claim 1 , caesium hydroxide (CsOH) or barium hydroxide (Ba(OH)).4. The process according to any one of to claim 1 , wherein the organic lysis solvent is a polar protic solvent.5. The process according to any of to claim 1 , wherein the organic lysis solvent is ethanol claim 1 , methanol claim 1 , n-butanol claim 1 , isopropanol or n-propanol.6. The process according to any one of to claim 1 , wherein the temperature is at least 30° C. above the boiling point of the lysis medium at atmospheric pressure.7. The process according to ...

ISOPRENE SYNTHASE AND GENE ENCODING THE SAME, AND METHOD FOR PRODUCING ISOPRENE MONOMER

The present invention provides means useful for establishing an excellent isoprene monomer production system. Specifically, the present invention provides a polynucleotide of the following (a), (b), or (c): 1. A polynucleotide of the following (a) , (b) , or (c):(a) a polynucleotide comprising (i) the nucleotide sequence represented by SEQ ID NO:1, or (ii) the nucleotide sequence consisting of the nucleotide residues at positions 133 to 1785 in the nucleotide sequence represented by SEQ ID NO:1;(b) a polynucleotide that comprises a nucleotide sequence having 90% or more identity to the nucleotide sequence of (i) or (ii), and encodes a protein having an isoprene synthase activity; or(c) a polynucleotide that hybridizes under a stringent condition with a polynucleotide consisting of the nucleotide sequence complementary to the nucleotide sequence of (i) or (ii), and encodes a protein having an isoprene synthase activity.2Mucuna.. The polynucleotide according to claim 1 , wherein the polynucleotide is derived from3. A protein of the following (A) claim 1 , (B) claim 1 , or (C):(A) a protein comprising (i′) the full length amino acid sequence represented by SEQ ID NO:2, or (ii′) the amino acid sequence consisting of the amino acid residues at positions 45 to 594 in the amino acid sequence represented by SEQ OD NO:2;(B) a protein that comprises an amino acid sequence having 90% or more identity to the amino acid sequence of (i′) or (ii′), and has an isoprene synthase activity; or(C) a protein that comprises an amino acid sequence having a deletion, substitution, addition or insertion of one or several amino acids in the amino acid sequence of (i′) or (ii′), and has an isoprene synthase activity.4. An expression vector comprising the polynucleotide according to .5. A transformant prepared by introducing the expression vector according to into a host.6. The transformant according to claim 5 , wherein the host has an ability to synthesize dimethylallyl diphosphate via a ...

MEMBRANE BIOREACTOR FOR INCREASED PRODUCTION OF ISOPRENE GAS

The invention provides improved methods for the production of isoprene from biological materials. 1124-. (canceled)125. A method of producing isoprene , the method comprising:(a) culturing cells comprising a heterologous nucleic acid encoding an isoprene synthase polypeptide under suitable culture conditions for the production of isoprene;(b) removing a portion of the culture;(c) filtering the removed portion of the culture to produce a permeate and a retentate;(d) returning the retentate to the culture; and(e) producing isoprene;wherein the cultured cells undergoing steps (b), (c), and (d) either produce isoprene at a higher titer, or have greater average volumetric productivity of isoprene than the same cells cultured without undergoing steps (b), (c), and (d). This application is a continuation of U.S. patent application Ser. No. 12/976,572, filed on Dec. 22, 2010, which claims priority to U.S. Provisional Patent Application No. 61/289,352, filed on Dec. 22, 2009, the disclosures of which are herein incorporated by reference in their entirety.This disclosure relates to improved methods for the production of isoprene.Isoprene (2-methyl-1,3-butadiene) is an important organic compound used in a wide array of applications. For instance, isoprene is employed as an intermediate or a starting material in the synthesis of numerous chemical compositions and polymers. Isoprene is also an important biological material that is synthesized naturally by many plants and animals, including humans.The isoprene used in industrial applications is typically produced as a by-product of the thermal cracking of petroleum or naphtha or is otherwise extracted from petrochemical streams. This is a relatively expensive, energy-intensive process. With the worldwide demand for petrochemical based products constantly increasing, the cost of isoprene is expected to rise to much higher levels in the long-term and its availability is limited in any case. There is concern that future supplies of ...

RECOMBINANT MICROORGANISMS FOR ENHANCED PRODUCTION OF MEVALONATE, ISOPRENE, AND ISOPRENOIDS

The invention features compositions and methods for the increased production of mevalonate, isoprene, isoprenoid precursor molecules, and/or isoprenoids in microorganisms by engineering a microorganism for increased carbon flux towards mevalonate production in the following enzymatic pathways: (a) citrate synthase, (b) phosphotransacetylase, (c) acetate kinase, (d) lactate dehydrogenase, (e) malic enzyme, and (f) pyruvate dehydrogenase such that one of more of the enzyme activity is modulated. In addition, production of mevalonate, isoprene, isoprenoid precursor molecules, and/or isoprenoids can be further enhanced by the heterologous expression of the mvaE and mvaS genes (such as, but not limited to, mvaE and mvaS genes from the organisms DSM 20601, EG2, and ). 2. The cells of claim 1 , wherein the one or more nucleic acids encoding MVA pathway polypeptides are from the upper MVA pathway claim 1 , wherein the upper MVA pathway nucleic acids are selected from the group consisting of AA-CoA thiolase claim 1 , HMG-CoA synthase claim 1 , and HMG-CoA reductase nucleic acids.3. The cells of claim 1 , wherein said one or more nucleic acids encoding an upper mevalonate (MVA) pathway polypeptide is an mvaE gene and an mvaS gene.4L. grayiE. faeciumE. gallinarumE. casseliflavusE. faecalis.. The cells of claim 3 , wherein the mvaE gene and the mvaS gene is selected from the group consisting of: (a) an mvaE gene and an mvaS gene from ; (b) an mvaE gene and an mvaS gene from ; (c) an mvaE gene and an mvaS gene from ; (d) an mvaE gene and an mvaS gene from ; and (e) an mvaE gene and an mvaS gene from5. The cells of claim 1 , wherein the one or more nucleic acids encoding MVA pathway polypeptides are from the lower MVA pathway claim 1 , wherein the lower MVA pathway nucleic acids are selected from the group consisting of MVK claim 1 , PMK claim 1 , and claim 1 , MVD nucleic acids.6M. mazeiM. burtoniiLactobacillusLactobacillus sakeiSaccharomyces cerevisiaeStreptococcusStreptococcus ...

MODIFIED MICROORGANISMS AND METHODS OF MAKING BUTADIENE USING SAME

The present disclosure generally relates to microorganisms that comprise one or more polynucleotides coding for enzymes in one or more pathways that catalyze a conversion of a fermentable carbon source to butadiene. Also provided are methods of using the microorganisms in industrial processes including, for use in the production of butadiene and products derived therefrom. 1. A method for producing butadiene , the method comprising: catalyzing a conversion of crotonyl alcohol to butadiene with an enzyme having an amino acid sequence at least 70% identical to linalool dehydratase (GI: 302064203).2. The method of claim 1 , wherein the enzyme is at least 80% identical to linalool dehydratase (GI: 302064203).3. The method of claim 1 , wherein the enzyme is at least 95% identical to linalool dehydratase (GI: 302064203).4. The method of claim 1 , wherein the enzyme is linalool dehydratase (GI: 302064203).5. The method of claim 1 , wherein the enzyme accepts crotonyl alcohol as a substrate.6. The method of claim 1 , wherein the enzyme has dehydratase activity.7. The method of claim 1 , wherein the enzyme has isomerase activity8. The method of claim 1 , wherein the enzyme has dehydratase and isomerase activity.9. The method of claim 1 , wherein the method is performed in a microorganism.10. A method for producing butadiene claim 1 , the method comprising: catalyzing a conversion of crotonyl alcohol to butadiene with an enzyme having an amino acid sequence at least 70% identical to a linalool dehydratase (EC 4.2.1.127).11. The method of claim 10 , wherein the enzyme is at least 80% identical to a linalool dehydratase (EC 4.2.1.127).12. The method of claim 10 , wherein the enzyme is at least 95% identical to a linalool dehydratase (EC 4.2.1.127).13. The method of claim 10 , wherein the enzyme is a linalool dehydratase (EC 4.2.1.127).14. The method of claim 10 , wherein the enzyme accepts crotonyl alcohol as a substrate.15. The method of claim 10 , wherein the enzyme has ...

Production of Alkenes by Combined Enzymatic Conversion of 3-Hydroxyalkanoic Acids

The present invention relates to a method for generating alkenes through a biological process. More specifically, the invention relates to a method for producing alkenes (for example propylene, ethylene, 1-butylene, isobutylene or isoamylene) from molecules of the 3-hydroxyalkanoate type. 119-. (canceled)20. A method for producing an alkene comprising the conversion of a 3-hydroxyalkanoate into said alkene by:(i) a first enzyme having an activity of converting the 3-hydroxyalkanoate into the corresponding 3-phosphonoxyalkanoate; and(ii) a second enzyme being different from the first enzyme and having an activity of converting said 3-phosphonoxyalkanoate into said alkene.21. The method of claim 20 , wherein the first enzyme is a mevalonate diphosphate (MDP) decarboxylase and the second enzyme is a different mevalonate diphosphate (MDP) decarboxylase.22. The method of wherein: (A) a protein comprising the amino acid sequence as shown in SEQ ID NO: 1 or a protein comprising an amino acid sequence which is at least 75% identical to the amino acid sequence shown in SEQ ID NO: 1 and showing an activity of converting the 3-hydroxyalkanoate into the corresponding 3-phosphonoxyalkanoate which is at least as high as the corresponding activity of the protein having the amino acid sequence shown in SEQ ID NO: 1;', '(B) a protein comprising the amino acid sequence as shown in SEQ ID NO: 2 or a protein comprising an amino acid sequence which is at least 75% identical to the amino acid sequence shown in SEQ ID NO: 2 and showing an activity of converting the 3-hydroxyalkanoate into the corresponding 3-phosphonoxyalkanoate which is at least as high as the corresponding activity of the protein having the amino acid sequence shown in SEQ ID NO: 2;', '(C) a protein comprising the amino acid sequence as shown in SEQ ID NO: 3 or a protein comprising an amino acid sequence which is at least 75% identical to the amino acid sequence shown in SEQ ID NO: 3 and showing an activity of ...

Polypeptide Assemblies and Methods for the Production Thereof

The application discloses multimeric assemblies including multiple oligomeric substructures, where each oligomeric substructure includes multiple proteins that self-interact around at least one axis of rotational symmetry, where each protein includes one or more polypeptide-polypeptide interface (“O interface”); and one or more polypeptide domain that is capable of effecting membrane scission and release of an enveloped multimeric assembly from a cell by recruiting the ESCRT machinery to the site of budding by binding to one or more proteins in the eukaryotic ESCRT complex (“L domain”); and where the multimeric assembly includes one or more subunits comprising one or more polypeptide domain that is capable of interacting with a lipid bilayer (“M domain”), as well as membrane-enveloped versions of the multimeric assemblies. 1. A multimeric assembly , comprising a plurality of oligomeric substructures , wherein each oligomeric substructure comprises a plurality of proteins that self-interact around at least one axis of rotational symmetry , wherein each protein comprises:(a) one or more polypeptide-polypeptide interface (“O interface”);(b) one or more polypeptide domain that is capable of effecting membrane scission and release of an enveloped multimeric assembly from a cell by recruiting the ESCRT machinery to the site of budding by binding directly or indirectly to one or more ESCRT or ESCRT-associated proteins (“L domain”);wherein the multimeric assembly comprises one or more polypeptide domain that is capable of interacting with a lipid bilayer (“M domain”);wherein the M domain, L domain, and O interface are not each present in a single naturally occurring protein, wherein the plurality of oligomeric substructures interact with each other at the one or more O interfaces.2. The multimeric assembly of claim 1 , wherein each oligomeric structure comprises one or more M domain claim 1 , or wherein each protein comprises one or more M domain.3. The multimeric assembly ...

HERBICIDE-RESISTANT RICE PLANTS, POLYNUCLEOTIDES ENCODING HERBICIDE-RESISTANT ACETOHYDROXYACID SYNTHASE LARGE SUBUNIT PROTEINS, AND METHODS OF USE

The present disclosure provides a method for treating rice. The method comprises the steps of: providing a domestic rice crop plant and at least one AHAS-inhibiting herbicide selected from the group comprising a sulfonylurea herbicide, a sulfonyl carboxamide herbicide, an imidazolinone herbicide, a triazolopyrimidine herbicide, a pyrimidinyloxybenzoate herbicide, and a sulfonylaminocarbonyltriazolinone herbicide; applying an effective amount (measured in g Al/Ha) of one or more of the aforementioned herbicide to the domestic rice crop plant, post-emergence; thereby creating a treated rice plant; and growing the resulting treated rice plant. 1. A method for treating rice , comprising:providing a rice crop plant and at least one sulfonylurea herbicide;applying an effective amount of the at least one sulfonylurea herbicide to the rice crop plant, post-emergence; thereby creating a treated rice plant; andgrowing the resulting treated rice plant.2. The method of claim 1 , further comprising harvesting seed from the treated rice plant.3. The method of or claim 1 , wherein the rice crop plant comprises an acetohydroxyacid synthase large subunit (AHASL) polynucleotide encoding a herbicide-tolerant AHASL polypeptide comprising a leucine substitution at a position corresponding to position 171 of SEQ ID NO:26.4. The method of any one of - claim 1 , wherein the rice crop plant comprises an AHASL polypeptide comprising a leucine substitution at the position corresponding to position 171 of SEQ ID NO:26.5. The method of any one of - claim 1 , wherein the rice crop plant comprises the herbicide tolerance characteristics of a plant of line IMINTA 15.6. The method of any one of - claim 1 , wherein the rice crop plant:(a) is a plant of line IMINTA 15;(b) is a progeny of a plant of line IMINTA 15;(c) is a mutant or recombinant plant of line IMINTA 15; or(d) is a progeny of any one of the plants of (a)-(c).7. The method of claim 1 , wherein the sulfonylurea herbicide comprises one or ...

ENGINEERED PHENYLALANINE AMMONIA LYASE POLYPEPTIDES

The present invention provides engineered phenylalanine ammonia-lyase (PAL) polypeptides and compositions thereof, as well as polynucleotides encoding the engineered phenylalanine ammonia-lyase (PAL) polypeptides. 1. An engineered polynucleotide encoding an engineered polypeptide comprising an amino acid sequence having at least 90% sequence identity to reference sequence SEQ ID NO:4 , wherein said amino acid sequence comprises an amino acid residue selected from leucine , methionine , and glutamine at position 305 , wherein said positions in said amino acid sequence are in reference to SEQ ID NO:4.2. The engineered polynucleotide encoding the engineered polypeptide of claim 1 , wherein said engineered polypeptide exhibits an improved property selected from reduced sensitivity to proteolysis claim 1 , increased tolerance to acidic pH claim 1 , reduced immunogenicity claim 1 , or a combination thereof claim 1 , as compared to the reference sequence SEQ ID NO:4.3. The engineered polynucleotide encoding the engineered polypeptide of claim 2 , wherein the improved property is selected from reduced sensitivity to proteolysis and/or increased tolerance to acidic pH.4. The engineered polynucleotide encoding the engineered polypeptide of claim 1 , wherein said engineered polypeptide is resistant to proteolysis claim 1 , acid stable claim 1 , and/or deimmunized.5. The engineered polynucleotide encoding the engineered polypeptide of claim 4 , wherein said engineered polypeptide is resistant to proteolysis by at least one digestive tract enzyme claim 4 , wherein said engineered polypeptide is resistant to proteolysis by chymotrypsin claim 4 , trypsin claim 4 , carboxypeptidases claim 4 , and/or elastases.6. The engineered polynucleotide encoding the engineered polypeptide of claim 5 , wherein said engineered polypeptide is deimmunized.7. The engineered polynucleotide encoding the engineered polypeptide of claim 1 , wherein said polypeptide is purified.8. The engineered ...

ENGINEERED PHENYLALANINE AMMONIA LYASE POLYPEPTIDES

The present invention provides engineered phenylalanine ammonia-lyase (PAL) polypeptides and compositions thereof, as well as polynucleotides encoding the engineered phenylalanine ammonia-lyase (PAL) polypeptides. 1. An engineered polypeptide comprising an amino acid sequence having at least 90% sequence identity to reference sequence SEQ ID NO:12 , wherein said amino acid sequence comprises an amino acid residue selected from leucine , methionine , and glutamine at position 305 , and wherein said engineered polypeptide further comprises at least one or more of the following substitutions or substitution sets selected from 18 , 18/47 , 18/47/214 , 18/47/214/308 , 18/47/214/308/450/546 , 18/47/214/450 , 18/47/214/467/540/546 , 18/47/214/540 , 18/47/214/546 , 18/47/308/546 , 18/47/450 , 18/47/450/528/546 , 18/47/450/546 , 18/214 , 18/214/308 , 18/214/308/450/467 , 18/214/450 , 18/214/460 , 18/214/467/546 , 18/214/540 , 18/450/540 , 18/540 , 47 , 47/214 , 47/214/308 , 47/214/540 , 47/214/546 , 47/450 , 59/134 , 59/134/240/304/353/521/564 , 59/134/240/353 , 59/134/240/521 , 59/134/240/521/564 , 59/134/240/521/565 , 59/134/304/353/521 , 59/134/353/509/521 , 59/134/509 , 59/240 , 59/240/353/509 , 59/240/353/564/565 , 59/240/509/521/564 , 59/353 , 59/353/509/564 , 59/353/521 , 59/353/565 , 59/509/564 , 59/521 , 59/564 , 134/240/304/353/564 , 134/240/353/509/521/564 , 134/240/353/521/564 , 134/240/509/521 , 134/304/353/521/565 , 134/353/509/564 , 134/353/521 , 134/353/521/564 , 134/353/564 , 134/521 , 214/308/460 , 214/450/528 , 214/540 , 214/546 , 240 , 240/304/353/509/521/564 , 240/304/509 , 240/304/509/521 , 240/353/509 , 240/353/509/521 , 240/353/509/521/564/565 , 240/353/521 , 240/509 , 240/509/564/565 , 240/509/565 , 240/521/564/565 , 304/353/509 , 304/509 , 308/450/467 , 353/521 , 521 , and 528/546 , wherein said positions in said amino acid sequence are in reference to SEQ ID NO:12.2. The engineered polypeptide of claim 1 , wherein said engineered polypeptide ...

ENGINEERED PHENYLALANINE AMMONIA LYASE POLYPEPTIDES

The present invention provides engineered phenylalanine ammonia-lyase (PAL) polypeptides and compositions thereof, as well as polynucleotides encoding the engineered phenylalanine ammonia-lyase (PAL) polypeptides. 1. An engineered polynucleotide encoding a polypeptide comprising an amino acid sequence with at least 90% sequence identity to reference sequence SEQ ID NO:24 , wherein said polypeptide comprises a mutation at position 509 , wherein said positions are numbered with reference to SEQ ID NO: 24.2. The engineered polynucleotide of claim 1 , wherein said encoded engineered polypeptide further comprises at least one amino acid residue differences as compared to SEQ ID NO:24 claim 1 , wherein the amino acid residue differences are at one or more amino acid positions selected from 20 claim 1 , 24 claim 1 , 27 claim 1 , 39 claim 1 , 43 claim 1 , 45 claim 1 , 47 claim 1 , 54 claim 1 , 58 claim 1 , 59 claim 1 , 62 claim 1 , 70 claim 1 , 73 claim 1 , 80 claim 1 , 82 claim 1 , 91 claim 1 , 94 claim 1 , 98 claim 1 , 104 claim 1 , 105 claim 1 , 110 claim 1 , 112 claim 1 , 115 claim 1 , 117 claim 1 , 118 claim 1 , 119 claim 1 , 121 claim 1 , 123 claim 1 , 124 claim 1 , 125 claim 1 , 126 claim 1 , 127 claim 1 , 128 claim 1 , 129 claim 1 , 130 claim 1 , 131 claim 1 , 133 claim 1 , 134 claim 1 , 135 claim 1 , 139 claim 1 , 140 claim 1 , 141 claim 1 , 142 claim 1 , 143 claim 1 , 144 claim 1 , 145 claim 1 , 146 claim 1 , 147 claim 1 , 149 claim 1 , 150 claim 1 , 151 claim 1 , 153 claim 1 , 154 claim 1 , 156 claim 1 , 157 claim 1 , 158 claim 1 , 159 claim 1 , 172 claim 1 , 174 claim 1 , 175 claim 1 , 176 claim 1 , 177 claim 1 , 178 claim 1 , 180 claim 1 , 187 claim 1 , 191 claim 1 , 195 claim 1 , 199 claim 1 , 205 claim 1 , 206 claim 1 , 210 claim 1 , 212 claim 1 , 213 claim 1 , 214 claim 1 , 232 claim 1 , 240 claim 1 , 243 claim 1 , 245 claim 1 , 247 claim 1 , 248 claim 1 , 250 claim 1 , 256 claim 1 , 257 claim 1 , 266 claim 1 , 270 claim 1 , 275 claim 1 , 278 claim 1 , 279 ...

ENGINEERED PHENYLALANINE AMMONIA LYASE POLYPEPTIDES

The present invention provides engineered phenylalanine ammonia-lyase (PAL) polypeptides and compositions thereof, as well as polynucleotides encoding the engineered phenylalanine ammonia-lyase (PAL) polypeptides. 1. An engineered polypeptide comprising an amino acid sequence with at least 90% sequence identity to reference sequence SEQ ID NO:24 , and wherein the engineered polypeptide comprises at least one amino acid residue differences as compared to SEQ ID NO:24 , wherein the amino acid residue differences are at one or more amino acid positions selected from 20 , 24 , 27 , 39 , 43 , 45 , 47 , 54 , 58 , 59 , 62 , 70 , 73 , 80 , 82 , 91 , 94 , 98 , 104 , 105 , 110 , 112 , 115 , 117 , 118 , 119 , 121 , 123 , 124 , 125 , 126 , 127 , 128 , 129 , 130 , 131 , 133 , 134 , 135 , 139 , 140 , 141 , 142 , 143 , 144 , 145 , 146 , 147 , 149 , 150 , 151 , 153 , 154 , 156 , 157 , 158 , 159 , 172 , 174 , 175 , 176 , 177 , 178 , 180 , 187 , 191 , 195 , 199 , 205 , 206 , 210 , 212 , 213 , 214 , 232 , 240 , 243 , 245 , 247 , 248 , 250 , 256 , 257 , 266 , 270 , 275 , 278 , 279 , 285 , 286 , 289 , 290 , 292 , 304 , 305 , 307 , 308 , 309 , 319 , 321 , 326 , 331 , 332 , 334 , 349 , 355 , 364 , 365 , 369 , 370 , 371 , 372 , 374 , 375 , 377 , 378 , 379 , 381 , 382 , 383 , 384 , 385 , 387 , 389 , 394 , 396 , 399 , 400 , 403 , 404 , 407 , 417 , 418 , 425 , 431 , 432 , 433 , 434 , 435 , 436 , 437 , 438 , 439 , 443 , 446 , 447 , 453 , 456 , 459 , 460 , 461 , 463 , 471 , 472 , 473 , 474 , 475 , 476 , 477 , 478 , 479 , 482 , 483 , 503 , 507 , 521 , 522 , 524 , 525 , 528 , 538 , 546 , 547 , 551 , 558 , 560 , 564 , 565 , and/or any combinations thereof , wherein the amino acid positions are numbered with reference to SEQ ID NO:24.2. The engineered polypeptide of claim 1 , wherein said amino acid residue difference is selected from one or more of the following substitutions 112C claim 1 , 134Q claim 1 , 158H claim 1 , 180A claim 1 , 195E claim 1 , 240R/W claim 1 , 243I/L claim 1 , 245L claim 1 , ...

METHODS FOR BIOSYNTHESIS OF ISOPRENE

This document describes biochemical pathways for producing isoprene by forming two vinyl groups in a central precursor produced from isobutyryl-CoA, 3-methyl-2-oxopentanoate, or 4-methyl-2-oxopentanoate as well as recombinant hosts for producing isoprene. 1. A method for enzymatically synthesizing isoprene , said method comprising enzymatically introducing a terminal vinyl group into 3-methyl-pent-2-enoyl-[acp] , 4-methyl-pent-2-enoyl-[acp] , 3-methyl-3-hydroxy-pentanoate , 4-methyl-3-hydroxypentanoate , or mevalonate , and converting the resulting product in one or more steps to isoprene.2. The method of claim 1 , wherein said first vinyl group is introduced using a dehydratase claim 1 , a monooxygenase claim 1 , a cytochrome P450 reductase claim 1 , or an acyl-[acp] dehydrogenase.3. The method of claim 1 , wherein said terminal vinyl group is introduced into 3-methyl-pent-2-enoyl-[acp] or 4-methyl-pent-2-enoyl-[acp] using an acyl-[acp] dehydrogenase.4. The method of claim 3 , where said acyl-[acp] dehydrogenase has at least 70% homology to the amino acid sequence set forth in SEQ ID NO:14.5. (canceled)6. The method of claim 2 , where the dehydratase is classified under EC 4.2.1.—and has at least 70% homology to the amino acid sequence set forth in SEQ ID NO: 22.7. The method of claim 1 , wherein said terminal vinyl group is introduced into 3-methyl-3-hydroxy-pentanoate or 4-methyl-3-hydroxypentanoate using a monooxygenase or a cytochrome P450 reductase.8. The method of claim 7 , wherein said monooxygenase has at least 70% homology to the amino acid sequence set forth in SEQ ID NO: 15.9. (canceled)10. A method for enzymatically synthesizing isoprene claim 7 , said method comprising enzymatically introducing a second terminal vinyl group into 3-methyl-3-hydroxy-pent-4-enoate claim 7 , 4-methyl-3-hydroxypent-4-enoate claim 7 , 4-methyl-3-sulphorylpent-4-enoyl-[acp] claim 7 , or 3-methyl-3-buten-2-ol to produce isoprene.11. The method of claim 10 , wherein said second ...

PRODUCTION OF MEVALONATE, ISOPRENE, AND ISOPRENOIDS USING GENES ENCODING POLYPEPTIDES HAVING THIOLASE, HMG-COA SYNTHASE AND HMG-COA REDUCTASE ENZYMATIC ACTIVITIES

The invention features compositions and methods for the increased production of mevalonate, isoprene, isoprenoid precursor molecules, and/or isoprenoids in microorganisms via the heterologous expression of the mvaE and mvaS genes from the organisms DSM 20601, EG2, and 145-. (canceled)47. The cells of claim 46 , wherein the mvaE gene and mvaS gene are from different organisms.48. The cells of claim 46 , wherein:{'i': L. grayi', 'E. faecium;, 'a) the mvaE gene is from and the mvaS gene is from'}{'i': L. grayi', 'E. gallinarum;, 'b) the mvaE gene is from and the mvaS gene is from'}{'i': L. grayi', 'E. casseliflavus;, 'c) the mvaE gene is from and the mvaS gene is from'}{'i': E. faecium', 'L. grayi;, 'd) the mvaE gene is from and the mvaS gene is from'}{'i': E. faecium', 'E. gallinarum;, 'e) the mvaE gene is from and the mvaS gene is from'}{'i': E. faecium', 'E. casseliflavus;, 'f) the mvaE gene is from and the mvaS gene is from'}{'i': E. gallinarum', 'L. grayi;, 'g) the mvaE gene is from and the mvaS gene is from'}{'i': E. gallinarum', 'E. faecium;, 'h) the mvaE gene is from and the mvaS gene is from'}{'i': E. gallinarum', 'E. casseliflavus;, 'i) the mvaE gene is from and the mvaS gene is from'}{'i': E. casseliflavus', 'L. grayi;, 'j) the mvaE gene is from and the mvaS gene is from'}{'i': E. casseliflavus', 'E. faecium, 'k) the mvaE gene is from and the mvaS gene is from ; or'}{'i': E. casseliflavus', 'E. gallinarum., 'l) the mvaE gene is from and the mvaS gene is from'}49Enterococcus faecalisL. grayi, E. faecium, E. gallinarumE. casseliflavus.. The cells of claim 46 , wherein the cells produce increased amounts of isoprene compared to isoprene-producing cells that (A) contain a mvaE gene from and (B) do not contain the mvaE gene and the mvaS gene from claim 46 , or50. The cells of claim 46 , wherein the nucleic acids encoding polypeptides of the lower MVA pathway comprise enzymes selected from: (a) an enzyme that phosphorylates mevalonate to mevalonate 5-phosphate; (b ...

ENGINEERED PHENYLALANINE AMMONIA LYASE POLYPEPTIDES

The present invention provides engineered phenylalanine ammonia-lyase (PAL) polypeptides and compositions thereof, as well as polynucleotides encoding the engineered phenylalanine ammonia-lyase (PAL) polypeptides. 1. An engineered polypeptide comprising an amino acid sequence having at least 90% sequence identity to reference sequence SEQ ID NO:4 , wherein said amino acid sequence comprises an amino acid residue selected from leucine , methionine , and glutamine at position 305 , wherein said positions in said amino acid sequence are in reference to SEQ ID NO:4.2. An engineered polypeptide comprising an amino acid sequence having at least 90% sequence identity to reference sequence SEQ ID NO:10 , wherein said amino acid sequence comprises an amino acid residue selected from leucine , methionine , and glutamine at position 305 , and wherein said engineered polypeptide further comprises at least one or more of the following substitution sets selected from 32/54/73/305/503/521/565 , 54/59/73/305/503/521/565 , and 54/59/305/503/521/565 , wherein said positions in said amino acid sequence are in reference to SEQ ID NO:10.3. An engineered polypeptide comprising an amino acid sequence having at least 90% sequence identity to reference sequence SEQ ID NO:12 , wherein said amino acid sequence comprises an amino acid residue selected from leucine , methionine , and glutamine at position 305 , and wherein said engineered polypeptide further comprises at least one or more of the following substitutions or substitution sets selected from 18 , 18/47 , 18/47/214 , 18/47/214/308 , 18/47/214/308/450/546 , 18/47/214/450 , 18/47/214/467/540/546 , 18/47/214/540 , 18/47/214/546 , 18/47/308/546 , 18/47/450 , 18/47/450/528/546 , 18/47/450/546 , 18/214 , 18/214/308 , 18/214/308/450/467 , 18/214/450 , 18/214/460 , 18/214/467/546 , 18/214/540 , 18/450/540 , 18/540 , 47 , 47/214 , 47/214/308 , 47/214/540 , 47/214/546 , 47/450 , 59/134 , 59/134/240/304/353/521/564 , 59/134/240/353 , 59/134/240 ...

THERMOSTABLE RUBISCO ACTIVASE COMPLEXES

The present invention relates to Rubisco activase (RCA) and functionally equivalent polypeptides that confer thermostability to a complex comprising Rubisco activase and Ribulose-1,5-bis-phosphate carboxylase/oxygenase (Rubisco). 1Oryza australiensisO. australiensis. A complex comprising a Rubisco activase (RCA) from and a Ribulose-1 ,5-bis-phosphate carboxylase/oxygenase (Rubisco) from a plant other than wherein the RCA confers thermostability to the complex.2. The complex of claim 1 , wherein the RCA comprises at least one of SEQ ID NOs: 2 to 37 claim 1 , 39 or 40.3. The complex of having at least 90% claim 2 , at least 95% claim 2 , at least 99% or 100% sequence identity to SEQ ID NOs: 39 or 40.4. The complex according to being formed in a modified or transgenic plant.5Oryza australiensisO. australiensis. Isolated RCA from claim 1 , or functional variants thereof claim 1 , capable of forming a complex with a Rubisco from a plant other than and conferring thermostability to the complex.6. The RCA of comprising at least one of SEQ ID NOs: 2 to 37 claim 5 , 39 or 40.7. The RCA of having at least 90% claim 5 , at least 95% claim 5 , at least 99% or 100% sequence identity to SEQ ID NO: 39 or 40.8. An isolated nucleic acid encoding the RCA of .9. The isolated nucleic acid according to comprising one or more of SEQ ID NOs: 41 claim 8 , 42 claim 8 , 50 claim 8 , 51 and 52.10. (canceled)11. The isolated nucleic acid according to having at least 90% claim 9 , at least 95% claim 9 , at least 99% or 100% sequence identity to SEQ ID NOs: 41 claim 9 , 42 claim 9 , 50 claim 9 , 51 or 52.12. A chimeric gene or vector comprising the nucleic acid of .13. The chimeric gene or vector of wherein the nucleic acid is operably linked to a promoter claim 12 , preferably a plant expressible promoter.14. The chimeric gene or vector of wherein the promoter is a heat-inducible promoter claim 13 , green-tissue promoter or a chemical-inducible promoter.15. A host cell comprising the complex of ...

ENGINEERED PHENYLALANINE AMMONIA LYASE POLYPEPTIDES

The present invention provides engineered phenylalanine ammonia lyase (PAL) polypeptides and compositions thereof, as well as polynucleotides encoding the engineered phenylalanine ammonia lyase (PAL) polypeptides. Methods for producing PAL enzymes are also provided. In some embodiments, the engineered PAL polypeptides are optimized to provide enhanced catalytic activities that are useful under industrial process conditions for the production of pharmaceutical compounds. 1. An engineered polynucleotide encoding an engineered phenylalanine ammonia lyase comprising a polypeptide sequence having at least 95% sequence identity to SEQ ID NO: 4 , wherein said engineered phenylalanine ammonia lyase comprises a substitution at position 104 and at least one substitution selected from positions 220 , 222 , and 359 in said polypeptide sequence , and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 8.2. An engineered polynucleotide sequence encoding at least one engineered phenylalanine ammonia lyase , wherein said polynucleotide sequence comprises at least 85% sequence identity to SEQ ID NO: 1 , 3 , 7 , 105 , 251 , 445 , 481 , 515 , 617 , 713 , 829 , 893 , 987 , and/or 1139 , wherein the polynucleotide sequence of said engineered phenylalanine ammonia lyase comprises at least one substitution at one or more positions.3. The engineered polynucleotide sequence of claim 1 , wherein said polynucleotide sequence comprises at least 85% sequence identity to SEQ ID NO: 1 claim 1 , 3 claim 1 , 7 claim 1 , 105 claim 1 , 251 claim 1 , 445 claim 1 , 481 claim 1 , 515 claim 1 , 617 claim 1 , 713 claim 1 , 829 claim 1 , 893 claim 1 , 987 claim 1 , and/or 1139.4. The engineered polynucleotide sequence of claim 1 , wherein said polynucleotide sequence comprises SEQ ID NO: 1 claim 1 , 3 claim 1 , 7 claim 1 , 105 claim 1 , 251 claim 1 , 445 claim 1 , 481 claim 1 , 515 claim 1 , 617 claim 1 , 713 claim 1 , 829 claim 1 , 893 claim 1 , 987 ...