SIMULTANEOUS SACCHARIFICATION AND CO-FERMENTATION OF GLUCOAMYLASE-EXPRESSING FUNGAL STRAINS WITH AN ETHANOLOGEN TO PRODUCE ALCOHOL FROM CORN

A conversion process provides using different co-cultured cell lines to express different sets of enzymes catalyzing the same process. For example, in a Simultaneous Saccharification and Co-Fermentation (SSCF) process, a starch substrate is converted to alcohol by contacting the substrate with yeast and Aspergillus niger cells. Because A. niger expresses an endogenous glucoamylase and alpha-amylase, these enzymes do not need to be added during the SSCF process.

Methods for obtaining corn oil from milled corn germ

Methods for obtaining corn oil from milled corn germ (e.g., dry milled corn germ), involving adding water, at least one acidic cellulase, at least one acidic protease, and at least one phytase to milled corn germ to obtain corn oil.

Enhanced Fermentation Process Using a Transglycosidase

The present invention relates to methods for improving fermentation processes, including increasing product yield, reduced viscosity, and/or reduced foaming. 1. A method of making a fermentation product comprising;fermenting a feedstock with a fermenting microorganism in a fermentation broth to make a fermentation product, wherein the fermenting comprises a transglycosidase, and wherein the fermentation product has a higher yield than a control fermentation lacking the transglycosidase.2. The method of further comprising recovering the fermentation product.3. The method of wherein the fermentation product is lysine claim 1 , lactic acid claim 1 , or glutamic acid.4Brevibacterium flavum, Corynebacterium glutamicumCorynebacterium glutamicumLactobacillus rhamnosus.. The method of wherein the fermenting organism is 10178 claim 1 , 10238 claim 1 , or5. The method of wherein the transglycosidase comprises SEQ ID NO:1 claim 1 , or a polypeptide have at least 80% claim 1 , at least 85% claim 1 , at least 90% claim 1 , at least 95% claim 1 , at least 98% claim 1 , or even at least 99% amino acid sequence identity to SEQ ID NO: 1.6Trichoderma.. The method of wherein the transglycosidase is produced in a7. The method of wherein the feedstock comprises starch liquifact claim 1 , granular starch claim 1 , pure glucose claim 1 , pure sucrose claim 1 , or sugar syrup.8. The method of further comprising foam reduction and/or viscosity reduction in the fermentation broth.9. The method of wherein the TrTG is present at 1-6 Kg/MT claim 1 , 2-5 Kg/MT claim 1 , or 2-4 Kg/MT.1012-. (canceled)13. A The method of claim 15 , wherein{'i': Trichoderma', 'Trichoderma', 'Trichoderma, 'the fermenting comprises a TrTG transglucosidase, wherein the lysine has a higher yield compared to a control fermentation lacking the transglucosidase, and wherein the fermentation broth has reduced viscosity compared to a control fermentation lacking the transglucosidase.'}14. A The method of claim 15 , wherein ...

BUTTIAUXELLA SP. PHYTASE VARIANTS

Provided herein are variants of sp. phytases that may be used in industrial applications including methods for starch liquefaction, alcohol fermentations and for enhancing phosphate digestion in foods and animal feeds. 1Buttiauxella. A phytase variant having improved thermal activity of at least about 5° C. as compared to wild type sp. phytase (SEQ ID NO: 1).2. The phytase variant of claim 1 , wherein the improved thermal activity is at least about 17° C. as compared to the phytase SEQ ID NO: 1.3. The phytase variant of or claim 1 , wherein the improved thermal activity is in a range from about 17 to about 21° C.4. The phytase variant of any one of to claim 1 , wherein the improved thermal activity is in a range from about 17.4 to about 20.2° C.5Buttiauxella. A thermostable phytase variant capable of retaining greater than 50% of its activity after exposure to an elevated temperature for 10 minutes at pH 5.5 in buffer claim 1 , wherein the elevated temperature is at least about 5° C. higher than a temperature at which wild type sp. phytase (SEQ ID NO: 1) retains greater than 50% of its activity.6. The thermostable phytase variant of claim 5 , wherein the elevated temperature is at least about 20.5° C. higher.7. The thermostable phytase variant of or claim 5 , wherein the elevated temperature is in a range from about 20.5° C. to about 27° C.8. The thermostable phytase variant of any one of to claim 5 , wherein the elevated temperature is in a range from about 20.2° C. to about 26.8° C.9Buttiauxella. A phytase variant having phytase activity and an amino acid sequence that varies from the amino acid sequence of wild type sp. phytase (SEQ ID NO: 1) claim 5 , wherein the amino acid sequence of the phytase variant comprises a variation at one or more positions corresponding to position 75 claim 5 , 76 claim 5 , 77 claim 5 , 198 claim 5 , 367 or 374 of SEQ ID NO: 1.10. The phytase variant of claim 9 , wherein the phytase variant further comprises one or more additional ...

Expression of granular starch hydrolyzing enzymes in trichoderma and process for producing glucose from granular starch sustrates

The present invention relates to filamentous fungal host cells and particularly Trichoderma host cells useful for the production of heterologous granular starch hydrolyzing enzymes having glucoamylase activity (GSHE). Further the invention relates to a method for producing a glucose syrup comprising contacting a granular starch slurry obtained from a granular starch substrate simultaneously with an alpha amylase and a GSHE at a temperature equal to or below the gelatinization temperature of the granular starch to obtain a composition of a glucose syrup.

Polypeptides having glucoamylase activity and method of producing the same

The present invention relates to polypeptides having glucoamylase activity with improved properties and to compositions comprising these polypeptides suitable for use in the production of a food, beverage (e.g. beer), feed, or biofuel. Also described is an improved and cost-effective process for isolating glucoamylases suitable for large scale protein purification procedures. Furthermore, different methods and uses related to glucoamylases according to the invention are disclosed, such as in a brewing process.

A food product comprising a low temperature rice protein concentrate

Rice protein concentrates prepared at low temperature exhibit improved functionality and beneficial physiological benefits, including lowered cholesterol and enhanced lactic acid dehydrogenase activity, without an increase in blood urea nitrogen. The rice protein concentration could be made into a wet dough with comparatively less water than a soy protein concentrate. Use of the rice protein concentrate thus improved processing steps in the formulation of a food article containing the concentrate. The food product advantageously shows an extending shelf life and improved palatable texture.

PRODUCTION OF ISOPRENE UNDER NEUTRAL pH CONDITIONS

Embodiments of the present disclosure relate to a process for producing isoprene 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 isoprene comprising culturing a host cell , which comprises a heterologous nucleic acid encoding an isoprene 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 variant has one amino acid modification compared to the parent glucoamylase.3. The method of claim 1 , wherein the HgGA is SEQ ID NO: 3.4Trichoderma reesei. The method of claim 3 , wherein the HgGA is produced from a host cell.5. The method of claim 1 , wherein the TrGA is SEQ ID No: 6.6. The method of claim 1 , wherein the RhGA is SEQ ID NO: 9.7. The method of claim 1 , wherein SSF are carried out at pH 6.0 to 7.5.8. The method of claim 1 , wherein SSF are carried out at pH 7.0 to 7.5.9. The method of claim 1 , wherein SSF is performed at a temperature in a range of about 30° C. to ...

TRICHODERMA REESEI ALPHA-AMYLASE IS A MALTOGENIC ENZYME

A maltogenic a-amylase from (TrAA) and variants thereof are useful in the production of high-maltose syrups from liquefied starch. Particularly high maltose concentrations are achieved when TrAA is used in the presence of a pullulanase. Expression hosts and encoding nucleic acids useful for producing TrAA and its variants also are provided. 1Trichoderma reesei. An isolated polypeptide comprising (i) residues 21-463 of SEQ ID NO:3 , or (ii) a variant of α-amylase (TrAA) , wherein the variant has α-amylase activity and at least 80% amino acid sequence identity to residues 21-463 of SEQ ID NO:3.2. A method of saccharifying liquefied starch to produce a maltose-rich syrup comprising: adding a polypeptide according to to a liquefied starch solution claim 1 , and saccharifying the liquefied starch solution claim 1 , wherein said saccharifying the liquefied starch solution produces a maltose-rich syrup.3. The method of claim 2 , wherein said polypeptide is added to the liquefied starch solution at about 0.3-1 kg per metric ton of dry solids.4. The method of claim 2 , wherein the liquefied starch solution is a slurry of liquefied starch at about 20-35% w/w dry solids.5. The method of claim 2 , wherein the liquefied starch solution is saccharified at about 50° C. to about 60° C.6. The method of claim 5 , wherein the liquefied starch solution is saccharified at about 55° C. to about 60° C.7. The method of claim 2 , wherein the liquefied starch solution is saccharified at about pH 4.0 to about pH 6.0.8. The method of claim 7 , wherein the liquefied starch solution is saccharified at about pH 4.2 to about pH 4.8.9. The method of claim 2 , further comprising a step of adding a pullulanase claim 2 , a β-amylase claim 2 , a fungal α-amylase that is not a TrAA claim 2 , a protease claim 2 , a cellulase claim 2 , a hemicellulase claim 2 , a lipase claim 2 , a cutinase claim 2 , an isoamylase claim 2 , or a combination thereof claim 2 , to the liquefied starch solution.10. The method ...

Endoglucanase For Reducing The Viscosity Of A Plant Materials Slurry

The present disclosure relates to composition comprising EG cellulase and methods of use, thereof. The compositions are useful, e.g., for reducing the viscosity of plant material slurry. 1. A method for reducing the viscosity of a plant material slurry comprising barley or oats , comprising adding to the slurry a composition comprising an isolated endoglucanase (EG) cellulase.2. The method of claim 1 , wherein the composition is substantially free of other cellulases.3. The method of claim 2 , wherein an addition cellulase is separately added to the slurry but is not required to reduce the viscosity of the slurry.45-. (canceled)6. The method of claim 1 , wherein an additional cellulase is separately added to the slurry but is not required to reduce the viscosity of the slurry.7. The method of claim 1 , wherein the EG cellulase is expressed in a filamentous fungus.8Trichoderma reesei.. The method of claim 7 , wherein the EG cellulase is expressed in9. The method of claim 8 , wherein the EG cellulase is expressed under control of the cbh1 promoter.10. The method of claim 1 , wherein the EG cellulase is purified to at least 70% of total protein in the composition.11. The method of claim 1 , wherein the EG cellulase is purified to at least 80% of total protein in the composition.12. The method of claim 1 , wherein the EG cellulase is purified to at least 90% of total protein in the composition.13. The method of claim 1 , wherein the EG cellulase is purified to at least 95% of total protein in the composition.14. The method of claim 1 , wherein the EG cellulase is purified to at least 97% of total protein in the composition.15Trichoderma reeseiHypocrea jecorina. The method of claim 1 , wherein the EG cellulase is a () EG cellulase.16. The method of claim 1 , wherein the reduction in viscosity as a result of adding the EG cellulase is at least equivalent to a reduction in viscosity as a result of adding a mixture of cellulases wherein EG is a component.17. The method of ...

ALPHA-AMYLASE BLEND FOR STARCH PROCESSING AND METHOD OF USE THEREOF

The present disclosure relates to an enzyme blend comprising a low pH, thermostable alpha-amylase and a alpha-amylase. The blend can include at least about 1.0 Liquefon Unit (LU) of the alpha-amylase for every 5.0 Modified Wohlgemuth Unit (MWU) of the low pH, thermostable alpha-amylase. The enzyme blend described is suitable for starch liquefaction and saccharification, ethanol production, and/or sweetener production, among other things. Also provided herein is a method of processing a starch by liquefying the starch with the low pH, thermostable alpha-amylase and the alpha-amylase, simultaneously or sequentially. 1Bacillus licheniformisB. licheniformis. An enzyme blend for processing a starch comprising a low pH , thermostable alpha-amylase and a alpha-amylase , wherein the low pH , thermostable alpha-amylase has an amino acid sequence that is at least about 80% identical to SEQ ID NO: 2 , and wherein the enzyme blend contains at least about 0.5 to about 5.0 Liquefon Units (LUs) of the alpha-amylase for every 5.0 Modified Wohlgemuth Units (MWUs) of the low pH , thermostable alpha-amylase.2B. licheniformis. The enzyme blend of claim 1 , wherein the alpha-amylase has an amino acid sequence that is at least about 70% identical to SEQ ID NO: 4.3B. licheniformisB. licheniformis. The enzyme blend of claim 2 , wherein the alpha-amylase is a variant having one or more altered properties compared to the alpha-amylase having a amino acid sequence of SEQ ID NO: 4.4. The enzyme blend of claim 3 , wherein the one or more altered properties include: substrate specificity claim 3 , substrate binding claim 3 , substrate cleavage pattern claim 3 , thermal stability claim 3 , pH activity profile claim 3 , pH stability profile claim 3 , stability towards oxidation claim 3 , stability at lower levers of calcium ion (Ca) claim 3 , specific activity claim 3 , or any combination thereof.5. The enzyme blend of claim 1 , wherein the low pH claim 1 , thermostable alpha-amylase comprises an ...

ALPHA-AMYLASE VARIANTS WITH ALTERED PROPERTIES

Disclosed are compositions comprising variants of alpha-amylase that have alpha-amylase activity and which exhibit altered properties relative to a parent AmyS-like alpha-amylase from which they are derived. The compositions comprise an additional enzyme such as a phytase. Also disclosed are methods of using the compositions, and kits related thereto. 149-. (canceled)50. An alpha-amylase variant comprising an amino acid sequence at least 95% identical to a parent hybrid alpha-amylase polypeptide comprising a combination of partial amino acid sequences deriving from at least two sequences selected from the group consisting of SEQ ID NO: 1 , SEQ ID NO: 2 , SEQ ID NO: 6 , SEQ ID NO: 7 , SEQ ID NO: 8 , SEQ ID NO: 9 , SEQ ID NO: 10 , SEQ ID NO: 11 , SEQ ID NO: 12 , SEQ ID NO: 15 , and SEQ ID NO: 16 , and having a substitution at an amino acid position corresponding to position 242 of SEQ ID NO: 1 ,{'sub': 'm', 'wherein said variant has alpha-amylase activity and a Tabout 5-10° C. higher than said wild-type alpha-amylase; and'}wherein the substitution at an amino acid position corresponding to position 242 of SEQ ID NO: 1 is selected from the group consisting of S242A, S242E, and S242Q.51. The alpha-amylase variant of claim 50 , wherein the amino acid sequence is at least 98% identical to the AmyS-like alpha-amylase polypeptide sequence of SEQ ID NO: 1.52. An isolated polynucleotide encoding an alpha-amylase variant comprising an amino acid sequence at least 95% identical to a parent hybrid alpha-amylase polypeptide comprising a combination of partial amino acid sequences deriving from at least two sequences selected from the group consisting of SEQ ID NO: 1 claim 50 , SEQ ID NO: 2 claim 50 , SEQ ID NO: 6 claim 50 , SEQ ID NO: 7 claim 50 , SEQ ID NO: 8 claim 50 , SEQ ID NO: 9 claim 50 , SEQ ID NO: 10 claim 50 , SEQ ID NO: 11 claim 50 , SEQ ID NO: 12 claim 50 , SEQ ID NO: 15 claim 50 , and SEQ ID NO: 16 claim 50 , and having a substitution at an amino acid position ...

TRICHODERMA REESEI ALPHA-AMYLASE ENHANCES SACCHARIFICATION OF CORN STARCH

A maltogenic α-amylase from (TrAA) and variants thereof in the presence of a glucoamylase are useful in the production of high-glucose syrups from liquefied starch, where the high-glucose syrups produced thereby contain at least about 97% glucose. In this process, TrAA advantageously suppresses the reversion of glucose to malto-oligosaccharides. Expression hosts and encoding nucleic acids useful for producing TrAA and its variants also are provided. 118-. (canceled)19Trichoderma reesei. A method of baking , comprising adding an isolated polypeptide comprising (i) residues 21-463 of SEQ ID NO:3 , or (ii) a variant of α-amylase (TrAA) , wherein the variant has α-amylase activity and at least 80% amino acid sequence identity to residues 21-463 of SEQ ID NO:3 to a substance to be baked , and baking the substance.20. (canceled) This application claims priority of U.S. Provisional applications 60/906,811, filed Mar. 14, 2007 and 60/906,812, filed Mar. 14, 2007, each of which is herein incorporated by reference in its entirety.Also attached is a sequence listing comprising SEQ ID NOS:1-7, which are herein incorporated by reference in their entirety.A maltogenic α-amylase from (TrAA), nucleic acids encoding the same, and host cells comprising the nucleic acids are provided. Methods of using TrAA include saccharification of starch to a glucose-rich syrup.High fructose corn syrup (HFCS) is a processed form of corn syrup having a high fructose content and a sweetness comparable to sugar, making HFCS useful as a sugar substitute in soft drinks and other processed foods. HFCS currently represents a billion dollar industry. The process of producing HFCS has progressed over the years from acid hydrolysis to a sequence of enzyme-catalyzed reactions:(1) Liquefaction: α-Amylases (EC 3.2.1.1) are first used to degrade a starch suspension containing 30-40% w/w dry solids (ds) to maltodextrans. α-Amylases are endohydrolases that catalyze the random cleavage of internal α-1,4-D- ...

COMPOSITIONS AND METHODS FOR GRAIN PROCESSING WITHOUT PH ADJUSTMENT

Described are compositions and methods relating to starch processing without a phytase pretreatment step and without adjustment of the slurry pH adjustment. 1. A method for performing starch liquefaction in a slurry comprising starch and phytate , the method comprising contacting the slurry with a thermostable phytase and an alpha-amylase under primary liquefaction or secondary liquefaction conditions , wherein the presence of the thermostable phytase increases the amount of starch liquefaction compared to an equivalent process in the absence of the phytase.2. The method of claim 1 , wherein the pH of the slurry is not adjusted before or after primary liquefaction or secondary liquefaction.3. The method of claim 1 , wherein the alpha-amylase is active at a pH lower than that at which it would be active in the absence of the phytase.4. The method of claim 1 , wherein the slurry does not require a phytase pretreatment step prior to primary liquefaction.5. The method of claim 1 , wherein the temperature of primary liquefaction and secondary liquefaction is 75° C. or higher.6. The method of claim 1 , wherein the temperature of primary liquefaction and secondary liquefaction is 80° C. or higher.7. The method of claim 1 , wherein the temperature of primary liquefaction and secondary liquefaction is 85° C. or higher.8. The method of claim 1 , wherein the temperature of secondary liquefaction is 90° C. or higher.9. The method of claim 1 , wherein the slurry does not require the addition of an anti-oxidant.10Buttiauxella. The method of claim 1 , wherein the phytase is obtained from a spp.11Buttiauxella. The method of claim 1 , wherein the phytase is a recombinant thermostable phytase derived from a spp. phytase.12. The method of claim 11 , wherein the phytase is selected from BP-110 (SEQ ID NO: 3) claim 11 , BP-111 (SEQ ID NO: 4) claim 11 , and BP-112 (SEQ ID NO: 5).13. The method of claim 12 , wherein the phytase is BP-111 (SEQ ID NO: 4).14. The method of claim 1 , wherein ...

Expression of Granular Starch Hydrolyzing Enzymes in Trichoderma and Process for Producing Glucose from Granular Starch Sustrates

The present invention relates to filamentous fungal host cells and particularly host cells useful for the production of heterologous granular starch hydrolyzing enzymes having glucoamylase activity (GSHE). Further the invention relates to a method for producing a glucose syrup comprising contacting a granular starch slurry obtained from a granular starch substrate simultaneously with an alpha amylase and a GSHE at a temperature equal to or below the gelatinization temperature of the granular starch to obtain a composition of a glucose syrup. 1Trichoderma reeseiTrichoderma reesei. A fermentation medium comprising a granular starch hydrolyzing enzyme having glucoamylase activity (GSHE) produced from a culture of , wherein the comprises a heterologous polynucleotide encoding a GSHE having at least 90% amino acid sequence identity with SEQ ID NO: 3 or with SEQ ID NO: 6.218-. (canceled) The present application is a divisional application of U.S. patent application Ser. No. 10/991,654, filed Nov. 18, 2004, and claims priority to U.S. Provisional Patent Application Ser. No. 60/524,279 entitled Expression of Granular Starch Hydrolyzing Enzyme in , filed Nov. 21, 2003; U.S. Provisional Patent Application Ser. No. 60/531,953 entitled Enzyme Compositions for Glucose Feed from Granular Starch Substrates, filed Dec. 22, 2003; and U.S. Provisional Patent Application Ser. No. 60/566,358 entitled Expression of Granular Starch Hydrolyzing Enzyme in , filed Apr. 28, 2004.The present invention relates to filamentous fungal host cells useful for the expression of heterologous granular starch hydrolyzing enzymes having glucoamylase activity (GSHE). The invention further relates to the use of the GSHE in methods for producing glucose syrup and other end products from granular starch substrates comprising contacting a granular starch substrate, at or below the gelatinization temperature of the granular starch, simultaneously with a starch liquefying amylase and a GSHE. The invention ...

FOOD PRODUCT COMPRISING A LOW TEMPERATURE RICE PROTEIN CONCENTRATE

Rice protein concentrates prepared at low temperature exhibit improved functionality and beneficial physiological benefits, including lowered cholesterol and enhanced lactic acid dehydrogenase activity, without an increase in blood urea nitrogen. The rice protein concentration could be made into a wet dough with comparatively less water than a soy protein concentrate. Use of the rice protein concentrate thus improved processing steps in the formulation of a food article containing the concentrate. The food product advantageously shows an extending shelf life and improved palatable texture. 2. The food product of claim 1 , wherein the food product is a food supplement or an ingredient of a food supplement.3. The food product of claim 1 , wherein the food product is a functional food or an ingredient of a functional food.4. The food product of claim 1 , wherein the food product is an animal feed additive or an ingredient of an animal feed additive.5. The food product of claim 1 , wherein the food product is food additive.6. A method of making a food product comprising a rice protein concentrate according to claim 1 , comprising:(a) hydrolyzing a substantial portion of starch in a rice substrate with (i) an enzyme having granular starch hydrolyzing (GSH) activity and (ii) a second starch hydrolyzing enzyme at a temperature at or below 72° C. and at a pH of 3.0 to 6.5 for a period of time sufficient to obtain a solubilized starch fraction and a residue fraction containing insoluble rice protein; and(b) separating the solubilized starch fraction from the residue fraction to obtain said rice protein concentrate.7. The method according to claim 6 , wherein the enzyme having GSH activity is a glucoamylase.8Humicola, RhizopusAspergillus.. The method according to claim 7 , wherein the glucoamylase is derived from a strain of claim 7 , or9. The method according to claim 6 , wherein the second starch hydrolyzing enzyme is an alpha-amylase.10. The method according to claim 9 , ...

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 , ...

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 ...

ALPHA-AMYLASE BLEND FOR STARCH PROCESSING AND METHOD OF USE THEREOF

The present disclosure relates to an enzyme blend comprising a low pH, thermostable alpha-amylase and a alpha-amylase. The blend can include at least about 1.0 Liquefon Unit (LU) of the alpha-amylase for every 5.0 Modified Wohlgemuth Unit (MWU) of the low pH, thermostable alpha-amylase. The enzyme blend described is suitable for starch liquefaction and saccharification, ethanol production, and/or sweetener production, among other things. Also provided herein is a method of processing a starch by liquefying the starch with the low pH, thermostable alpha-amylase and the alpha-amylase, simultaneously or sequentially. 1Bacillus licheniformisB. licheniformisB. licheniformis. An enzyme blend for processing a starch comprising a low pH , thermostable alpha-amylase and a alpha-amylase , wherein the low pH , thermostable alpha-amylase has an amino acid sequence that is at least about 80% identical to SEQ ID NO: 2 , wherein the alpha-amylase has an amino acid sequence that is at least about 80% identical to SEQ ID NO: 6 , and wherein the enzyme blend contains at least about 0.5 to about 5.0 Liquefon Units (LUs) of the alpha-amylase for every 5.0 Modified Wohlgemuth Units (MWUs) of the low pH , thermostable alpha-amylase.23-. (canceled)4B. licheniformisB. licheniformis. The enzyme blend of claim 1 , wherein the alpha-amylase is a variant having one or more altered properties compared to the alpha-amylase having an amino acid sequence of SEQ ID NO: 6 claim 1 , wherein the one or more altered properties include: substrate specificity claim 1 , substrate binding claim 1 , substrate cleavage pattern claim 1 , thermal stability claim 1 , pH activity profile claim 1 , pH stability profile claim 1 , stability towards oxidation claim 1 , stability at lower levers of calcium ion (Ca2+) claim 1 , specific activity claim 1 , or any combination thereof.5. The enzyme blend of claim 1 , wherein the low pH claim 1 , thermostable alpha-amylase comprises an amino acid sequence of SEQ ID NO: 2. ...

SINGLE PH PROCESS FOR STARCH LIQUEFACTION AND SACCHARIFICATION FOR HIGH-DENSITY GLUCOSE SYRUPS

Embodiments of the present disclosure relate to a process for producing downstream products, such as high-density glucose syrups and high-glucose fermentation feedstock, from a starch-containing substrate without a pH adjustment before the saccharification step. The saccharification is effectively catalyzed by a glucoamylase at a pH in the range of 5.2 to 5.6. 1Humicola grisea. A method of processing a starch comprising saccharification of a starch substrate to glucose in the presence of a glucoamylase without a pH adjustment , wherein saccharification is performed at pH 5.2 to 5.6 and at a temperature in the range of about 58° C. to about 62° C. , wherein the glucoamylase is selected from the group consisting of a parent glucoamylase (HgGA) comprising SEQ ID NO: 3 and a variant thereof , wherein the variant has at least 99% identity to the parent glucoamylase.2. The method of claim 1 , wherein saccharification is performed in the presence of a pullulanase.3. The method of claim 1 , wherein the variant glucoamylase has at least one amino acid modification compared to the parent glucoamylase.4. The method of claim 1 , wherein the glucoamylase is HgGA having amino acid sequence of SEQ ID NO: 3.5Trichoderma reesei. The method of claim 1 , wherein the glucoamylase is produced in a host cell.6. The method of claim 1 , wherein the glucoamylase is added at a dose of at least about 0.15 GAU per gram of dry substance starch.7. The method of claim 1 , wherein the glucoamylase is added at a dose of at least about 0.20 GAU per gram of dry substance starch.8. The method of claim 1 , wherein saccharification is carried out at pH 5.35 to 5.5.9. The method of claim 1 , wherein the starch substrate is liquefied starch.10. The method of claim 1 , wherein saccharification achieves a DP1 concentration of at least 95% at about 70 hours.11. The method of claim 1 , wherein the starch substrate is about 30% to about 50% dry solids (DS).12. The method of claim 11 , wherein the starch ...

Methods for producing end-products from carbon substrates

The present invention provides means for the production of desired end-products of in vitro and/or in vivo bioconversion of biomass-based feed stock substrates, including but not limited to such materials as starch and cellulose. In particularly preferred embodiments, the methods of the present invention do not require gelatinization and/or liquefaction of the substrate.

PROCESS FOR PRODUCTION OF BIO-ALCOHOL

A process of enhanced bio-alcohol production from molasses is provided herein. The process comprises contacting molasses with sodium benzoate, potassium sorbate or a mixture thereof followed by yeast fermentation resulting in at least 6% increased alcohol production. Also included within the scope of the present invention is a process for producing co-product containing reduced glycerol. 1. A process for enhanced production of alcohol by fermentation of molasses , comprising contacting the molasses with at least one organic acid salt ranging from 10 ppm to 1000 ppm and fermenting said molasses in the presence of a fermenting organism , wherein said organic acid salt is sodium benzoate , or potassium sorbate , wherein said contacting occurs as a pre-treatment.2. The process of claim 1 , wherein said alcohol is selected from a group consisting of ethanol claim 1 , butanol claim 1 , methanol claim 1 , and propanol.3. The process of claim 1 , wherein said fermenting organism is a recombinant organism.4. The process of claim 1 , wherein said fermenting organism is a yeast.5. The process of claim 1 , wherein said yeast is a recombinant yeast.6SaccharomycesSaccharomyces cerevisiae, CandidaPichiaDekkeraHanseniasporaPseudozymaSacharromycodesZygosaccharomycesZygosaccharomyces rouxii, ZygoascusIssatchenkiaWilliopsisTorulaspora delbrueckii, Debaryomyces hansenii, Zygosaccharomyces bailii, BrettanomycesBrettanomyces bruxellensis.. The process of claim 5 , wherein said yeast is a sp. claim 5 , sp. claim 5 , sp. claim 5 , sp. claim 5 , sp. claim 5 , sp. claim 5 , sp. claim 5 , sp. claim 5 , sp. claim 5 , sp. claim 5 , sp. claim 5 , sp. and7. (canceled)8. A composition comprising yeast enriched with at least one organic acid salt or a mixture thereof.9. The composition as claimed in claim 8 , wherein said organic acid salt is sodium benzoate or potassium sorbate or a mixture thereof.10. The composition as claimed in claim 8 , wherein said composition is in form of yeast cake claim ...

LIQUEFACTION AND SACCHARIFICATION OF GRANULAR STARCH AT HIGH CONCENTRATION

The present teachings provide methods of processing granular starch in slurries containing high dry solids content. The slurries are initially incubated with enzymes at or below the gelatinization temperature. The use of pullulanase and glucoamylase at specified doses allows for improved glucose yields at lower energy cost. 1. A method of processing granular starch comprising:(a) contacting granular starch, water and one or more granular starch hydrolyzing enzymes including an alpha-amylase and/or a glucoamylase to produce slurry in which the concentration of dry solids is greater than 38% by weight,(b) incubating the slurry at a temperature above 40° C. and at or below the gelatinization temperature of the granular starch for at least five minutes to produce a composition in which the granular starch has been partially hydrolyzed into oligo- and/or mono-saccharides by the one or more enzymes; and(c) raising and holding the temperature of the partially hydrolyzed composition above the gelatinization temperature of the granular starch to produce a liquefied composition.2. The method of claim 1 , further comprising (d) contacting the liquefied composition with pullulanase and glucoamylase and incubating to produce glucose.34-. (canceled)5. The method of claim 1 , wherein the percentage of dry solids is at least 39% throughout steps (a)-(d).6. (canceled)7. The method of claim 1 , wherein the percentage of dry solids remains the same or increases between steps (a) and (d).8. The method of claim 2 , wherein no more than 10% by weight water is added to the slurry throughout steps (b) claim 2 , (c) and (d).9. The method of claim 2 , wherein no more water is added to the slurry throughout steps (b) claim 2 , (c) and (d).10. The method of claim 1 , wherein the one or more enzymes includes one or more alpha amylase.11. The method of claim 10 , wherein the alpha amylase is an alpha amylase selected from the group consisting of SPEZYME® AA claim 10 , SPEZYME® XTRA® claim 10 , ...

TRICHODERMA REESEI HOST CELLS EXPRESSING A GLUCOAMYLASE FROM ASPERGILLUS FUMIGATUS AND METHODS OF USE THEREOF

Fungal glucoamylases from —expressed in host cells (AfGATR) are provided. host cells express AfGATRs at higher, or at least comparable, levels to natively expressed AfGA . AfGATRs, including AfGA1TR and AfGA2TR, exhibit high activity at elevated temperatures and at low pH, so AfGATRs can be used efficiently in a process of saccharification in the presence of alpha-amylase, such as alpha-amylase (AkAA). AfGATRs advantageously catalyze starch saccharification to an oligosaccharide composition significantly enriched in DP1 (i.e., glucose) compared to the products of saccharification catalyzed by glucoamylase (AnGA) or native AfGA expressed in . AfGATRs such as AfGA1TR, AfGA2TR or a variant thereof can be used at a lower dosage than AnGA and natively expressed AfGAs to produce comparable levels of glucose. 129-. (canceled)30. A yeast cell that expresses AfGA or variant thereof , wherein the AfGA or variant thereof comprises an amino acid sequence with at least 90% sequence identity to the sequence of SEQ ID NO: 12 or 13.31. The yeast cell according to claim 30 , wherein the yeast cell secretes the AfGA.32. The yeast cell according to claim 30 , wherein the AfGA or variant thereof comprises an amino acid sequence with at least 95% or 99% sequence identity to the sequence of SEQ ID NO: 12 or 13.33. The yeast cell according to claim 32 , wherein the AfGA comprises the sequence of SEQ ID NO: 12 or 13.34. The yeast cell according to claim 30 , wherein the yeast cell further expresses an additional glucoamylase.35. Use of a yeast cell that expresses AfGA or variant thereof claim 30 , in a simultaneous saccharification and fermentation (SSF) process claim 30 , wherein the AfGA or variant thereof comprises an amino acid sequence with at least 80% sequence identity to SEQ ID NO: 12 or 13 claim 30 , and wherein the yeast cell and the AfGA or variant thereof are present in the same process step.36. The use according to claim 35 , wherein the AfGA or variant thereof comprises an ...

ENZYMATIC HYDROLYSIS OF DISACCHARIDES AND OLIGOSACCHARIDES USING ALPHA-GLUCOSIDASE ENZYMES

A method is disclosed for hydrolyzing an alpha-1,3 or alpha-1,6 glucosyl-glucose linkage in a saccharide (disaccharide or oligosaccharide). This method comprises contacting the saccharide with an alpha-glucosidase enzyme such as transglucosidase under suitable conditions, during which contacting step the enzyme hydrolyzes at least one alpha-1,3 or alpha-1,6 glucosyl-glucose linkage of the saccharide. This method is useful for reducing the amount of oligosaccharides in a filtrate isolated from a glucan synthesis reaction, for example. 115-. (canceled)16. A method that comprises:(a) providing an alpha-glucan synthesis reaction that comprises a soluble disaccharide or oligosaccharide byproduct, wherein at least 90% of the glycosidic linkages of the alpha-glucan are alpha-1,3-glycosidic linkages; and(b) contacting the soluble disaccharide or oligosaccharide byproduct with an alpha-glucosidase enzyme, wherein the amount of the soluble disaccharide or oligosaccharide byproduct is reduced compared to the amount of the soluble disaccharide or oligosaccharide byproduct that was present prior to said contacting.17. The method of claim 16 , wherein the alpha-glucosidase enzyme is immobilized.18. The method of claim 16 , wherein the degree of polymerization of the oligosaccharide byproduct prior to said contacting with the alpha-glucosidase is 3 to 7.19. The method of claim 16 , wherein said contacting of the soluble disaccharide or oligosaccharide byproduct with the alpha-glucosidase enzyme is performed in the alpha-glucan synthesis reaction.20. The method of claim 16 , further comprising obtaining a fraction of the alpha-glucan synthesis reaction that comprises the soluble disaccharide or oligosaccharide byproduct claim 16 , wherein said contacting of the soluble disaccharide or oligosaccharide byproduct with the alpha-glucosidase enzyme is performed in the fraction.21. The method of claim 20 , wherein the fraction is a filtrate of the alpha-glucan synthesis reaction.22. The ...

Methods For Obtaining Oil From Maize Using Acid Protease and Cell-wall Polysaccharide-degrading Enzymes

Disclosed are methods for obtaining oil from maize, involving grinding maize kernels to form flour, adding water to the flour to form a slurry, and incubating the slurry with α-amylase for about 10 minutes to about 180 minutes at a temperature of about 75° to about 120° C. and at a pH of about 3 to about 7 to form a mash, cooling the mash to about 15° C. to about 40° C. and adding a nitrogen source, glucoamylase, yeast, acid protease, and cell-wall polysaccharide-degrading enzymes to form a beer containing ethanol and oil, wherein the beer has a pH of about 3 to about 7, and recovering oil from the beer. 1. A method for obtaining oil from maize , comprising grinding maize kernels to form flour , adding water to said flour to form a slurry , and incubating said slurry with α-amylase for about 10 minutes to about 180 minutes at a temperature of about 75° to about 120° C. and at a pH of about 3 to about 7 to form a mash , cooling said mash to about 15° C. to about 40° C. and adding a nitrogen source , glucoamylase , yeast , acid protease , and cell-wall polysaccharide-degrading enzymes to form a beer containing ethanol and oil , wherein said beer has a pH of about 3 to about 7 , and recovering oil from said beer.2. The method according to claim 1 , wherein said cell-wall polysaccharide-degrading enzymes are cellulases and hemicellulases.3. The method according to claim 1 , wherein the concentration of said acid protease and cell-wall polysaccharide-degrading enzymes is about 0.25 kg to about 15 kg per metric ton of corn on a dry weight basis.4. The method according to claim 1 , wherein the concentration of said acid protease and cell-wall polysaccharide-degrading enzymes is about 0.5 to about 10 kg per metric ton of corn.5. The method according to claim 1 , wherein the concentration of said acid protease and cell-wall polysaccharide-degrading enzymes is about 0.5 to about 5 kg per metric ton of corn.6. The method according to claim 1 , wherein said flour has particle ...

Process for conversion of granular starch to ethanol

The present invention concerns a method of producing glucose from a granular starch substrate comprising, contacting a slurry comprising granular starch obtained from plant material with an alpha-amylase at a temperature below the starch gelatinization temperature of the granular starch to produce oligosaccharides and hydrolyzing the oligosaccharides to produce a mash comprising at least 20% glucose and further comprising fermenting the mash to obtain ethanol.

Method for Making High Maltose Syrup

The present teachings provide direct conversion of granular starch into a soluble sugar composition comprising a high, very high, and/or ultra high maltose content. The method involves contacting an aqueous slurry of granular starch with an enzyme composition comprising an appropriate ratio of an alpha-amylase and a maltogenic enzyme to produce a soluble starch substrate that is enzymatically converted to a sugar composition containing higher maltose. The process may use more than one temperature to achieve the same. 1. A method of making a high DP2 syrup containing at least 50% DP2 comprising;solubilizing a granular starch substrate at or below the initial gelatinization temperature with an exogenous alpha-amylase to form a mixture comprising dextrins;hydrolyzing the dextrins with a maltogenic enzyme to form the high DP2 syrup, wherein the ratio of alpha-amylase dose expressed as AAU/gds, to maltogenic enzyme dose expressed as DP degrees, is less than 8.23-. (canceled)4. The method according to further comprising a debranching enzyme5. (canceled)6. The method according to wherein the starch is whole ground grain selected from the group consisting of corn claim 1 , wheat claim 1 , barley claim 1 , rye claim 1 , triticale claim 1 , rice claim 1 , oat claim 1 , beans claim 1 , banana claim 1 , potato claim 1 , sweet potato claim 1 , sorghum claim 1 , legumes claim 1 , cassava claim 1 , millet claim 1 , potato claim 1 , or tapioca.712-. (canceled)13. The method according to wherein the maltogenic enzyme is selected from the group consisting of a barley beta-amylase OPTIMALT® BBA or Betalase 1500 claim 1 , a wheat beta-amylase claim 1 , a soy beta-amylase β-amylase#1500S claim 1 , a fungal alpha-amylase CLARASE® L claim 1 , maltogenic amylase is MAX-LIFE™ P100 or Maltogenase L.1419-. (canceled)20Bacilluspaenibacillus polymyxa, Thermoanaerobacterium thermosulfurigenes, Xanthophyllomyces dendrorhous, Bacillus cereus, Bacillus megaterium, Arabidopsis thalianaAspergillusA. ...

Process for producing high glucose compositions by simultaneous liquefaction and saccharification of starch substrates

Fungal glucoamylases from Aspergillus fumigatus that are expressed in Trichoderma reesei host cells (AfGATR) are provided. AfGATRs, including AfGA1TR and AfGA2TR, exhibit high activity at elevated temperatures and at low pH, so AfGATRs can be used efficiently in a simultaneous liquefaction and saccharification process in the presence of alpha amylase, such as Aspergillus kawachii alpha-amylase (AkAA). This greatly reduces the combined run time of liquefaction and saccharification reaction, where the pH and temperature must be readjusted for optimal use of the alpha-amylase or glucoamylase.

TRICHODERMA REESEI HOST CELLS EXPRESSING A GLUCOAMYLASE FROM ASPERGILLUS FUMIGATUS AND METHODS OF USE THEREOF

Fungal glucoamylases from —expressed in host cells (AfGATR) are provided. host cells express AfGATRs at higher, or at least comparable, levels to natively expressed AfGA . AfGATRs, including AfGA1TR and AfGA2TR, exhibit high activity at elevated temperatures and at low pH, so AfGATRs can be used efficiently in a process of saccharification in the presence of alpha-amylase, such as alpha-amylase (AkAA). AfGATRs advantageously catalyze starch saccharification to an oligosaccharide composition significantly enriched in DP1 (i.e., glucose) compared to the products of saccharification catalyzed by glucoamylase (AnGA) or native AfGA expressed in . AfGATRs such as AfGA1TR, AfGA2TR or a variant thereof can be used at a lower dosage than AnGA and natively expressed AfGAs to produce comparable levels of glucose. 1Trichoderma reeseiTrichoderma reeseiAspergillus fumigatus. A recombinant host cell expressing an AfGATR , or variant thereof , having at least 80% sequence identity to SEQ ID NO: 12 or 13 , wherein said host cell expresses the AfGATR or variant at a comparable level to a host cell that expresses an AfGA , or variant thereof , having the same amino acid sequence of the AfGATR , or variant thereof , under identical conditions.2Trichoderma reeseiA. fumigatus. A recombinant host cell expressing an AfGATR , or variant thereof , having at least 80% sequence identity to SEQ ID NO: 12 or 13 , wherein the AfGATR , or variant thereof , is more thermostable than an AfGA , or variant thereof , having the same amino acid sequence of AfGATR , or variant thereof , and wherein the AfGA , or variant thereof , is expressed in an host cell.3. A recombinant AfGATR claim 2 , or variant thereof claim 2 , produced by the host cell of .4. A recombinant AfGATR claim 1 , or variant thereof claim 1 , produced by the host cell of .5. The recombinant AfGATR claim 4 , or variant thereof claim 4 , of claim 4 , wherein said AfGATR claim 4 , or variant thereof claim 4 , has at least 70% activity at ...

AMYLASE WITH MALTOGENIC PROPERTIES

The present teachings provide an amylase with maltogenic properties. Nucleic acids encoding the maltogenic amylase and variants thereof, expression vectors, formulations, and host cells are also provided. Additional embodiments of the present teachings provide various methods of use and methods of manufacturing. 1. An isolated nucleic acid comprising the nucleotide sequence of SEQ ID NO: 1 or of a degenerate variant of SEQ ID NO: 1.2. An isolated nucleic acid comprising a sequence that encodes a polypeptide consisting of the amino acid sequence of SEQ ID NO: 3.3. An isolated nucleic acid comprising a sequence that hybridizes under stringent conditions to a hybridization probe the nucleotide sequence of which consists of SEQ ID NO: 1 , or the complement of SEQ ID NO: 1.4. An isolated nucleic acid comprising a sequence at least 66% , 67% , 68% , 69% , 70% , 75% , 80% , 85% , 90% , 95% , 98% , 99% , or 99.5% identical to SEQ ID NO: 1.5. The isolated nucleic acid of wherein the nucleic acid encodes a polypeptide that has starch hydrolysis activity.6. An isolated nucleic acid comprising a sequence that encodes a polypeptide at least 66% claim 4 , 67% claim 4 , 68% claim 4 , 69% claim 4 , 70% claim 4 , 75% claim 4 , 80% claim 4 , 85% claim 4 , 90% claim 4 , 95% claim 4 , 98% claim 4 , 99% claim 4 , or 99.5% identical to SEQ ID NO: 3 claim 4 , wherein the polypeptide has starch hydrolysis activity.7. An isolated nucleic acid comprising a sequence that encodes a polypeptide comprising the sequence of SEQ ID NO: 3 claim 4 , or SEQ ID NO: 3 with up to 50 conservative amino acid substitutions claim 4 , wherein the polypeptide has starch hydrolysis activity.8. A purified polypeptide claim 4 , the amino acid sequence of which comprises a sequence at least 66% claim 4 , 67% claim 4 , 68% claim 4 , 69% claim 4 , 70% claim 4 , 75% claim 4 , 80% claim 4 , 85% claim 4 , 90% claim 4 , 95% claim 4 , 98% claim 4 , 99% claim 4 , or 99.5% identical to SEQ ID NO: 3.9. A purified ...

Buttiauxella sp. phytase variants

Provided herein are variants of Buttiauxella sp. phytases that may be used in industrial applications including methods for starch liquefaction, alcohol fermentations and for enhancing phosphate digestion in foods and animal feeds.

ENHANCED FERMENTATION PROCESS USING MOLASSES

Described herein are compositions and methods for enhanced fermentation of molasses using transglucosidase. 1. A method for increasing the fermentation yield of molasses in a fermentation medium comprising contacting the molasses with an enzyme composition comprising at least one transglucosidase capable of hydrolyzing non-fermentable sugars such as raffinose and/or stachyose.2. The method of claim 1 , further comprising the addition of one or more secondary enzyme to hydrolyze granular starch claim 1 , proteins and/or residual starch in the molasses.3. The method of claim 1 , wherein fermentable sugars from molasses in the fermentation reaction are converted to obtain end products claim 1 , such as an alcohol claim 1 , an organic acid or a specialty biochemical.4. The method of claim 3 , wherein the alcohol is ethanol.5. The method of claim 3 , wherein the organic acid is lactic acid or citric acid.6. The method of claim 3 , wherein the specialty biochemical is an amino acid.7. A method for increasing the fermentable sugars in molasses comprising contacting molasses with at least one transglucosidase enzyme to obtain fermentable sugars.8. The method of claim 7 , further comprising fermenting the fermentable sugars to end products in the presence of fermenting microorganisms.9. The method of claim 8 , wherein the contacting and fermenting occur simultaneously or wherein the contacting is a pretreatment.10. The method of claim 8 , wherein the molasses comprises non-fermentable starch and the contacting claim 8 , pretreatment and/or fermenting occurs at a temperature below the gelatinization temperature of the granular starch.11. The method of claim 10 , wherein the non-fermentable starch is granular starch.12. The method of claim 10 , wherein the contacting and/or pretreatment temperature is below about 52° C.13. The method of claim 10 , wherein the fermentation temperature is from about 15 to 40° C.14. The method of claim 10 , wherein the fermentation claim 10 , ...

HIGH FORCE AND HIGH STRESS DESTRUCTURING FOR STARCH BIOMASS PROCESSING

A process for mechanical destructuring of starch-based biomass was developed that makes use of a short application of high compression, impact, and shearing forces. The biomass may be destructured using a specific energy input that is less than 40% of the total combustible energy of the biomass. The destructured starch-based biomass, with or without saccharification and/or in-feed glycosyl hydrolase enzymes, may be used in feed applications. The destructured starch-based may saccharified to produce syrups and fermentable sugars, and for production of products including ethanol using a biocatalyst. 1. A process for producing destructured starch-based biomass comprising:a) providing a starch-based biomass; andb) applying to the biomass of (a) at least one set of compression and impact forces of at least 5,000 N combined with shearing forces;wherein contact stress of greater than 5,000 psi is applied to the biomass and wherein a destructured starch-based biomass is produced.2. A process for producing destructured starch-based biomass comprising:a) providing a portion of starch-based biomass; andb) applying to the biomass of (a) at least one set of compression and impact forces of at least 1,500 N combined with shearing forces, and contact stress of greater than 5,000 psi;wherein specific energy input in the process is less than 40% of the total combustible energy of the portion of biomass being treated and wherein a destructured starch-based biomass is produced.3. The process of wherein the compression and impact forces are at least 3 claim 2 ,000 N.4. The process of or wherein the stress is greater than about 8 claim 2 ,000 psi.5. The process of wherein the stress is greater than about 13 claim 4 ,000 psi.6. The process of wherein the forces are imparted using a non-vibratory apparatus which does not contain free flowing media.7. The process of or wherein the forces are imparted by centrifugal motion claim 1 , hydraulics claim 1 , or springs.8. The process of wherein ...

DIRECT STARCH TO FERMENTABLE SUGAR

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 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 of making a fermentable sugar feedstock , the method comprising treating an aqueous slurry of ground or fractionated grain with an alpha amylase and a glucoamylase to produce the fermentable sugar feedstock;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 about 20 AAU/gds; andwherein the fermentable sugar feedstock comprises a higher concentration of DP-2 saccharides in comparison to fermentable sugar feedstocks that are not made by treating an aqueous slurry of ground or fractionated grain with a starch solubilizing alpha amylase and a glucoamylase, wherein the treatment is at a temperature below the initial gelatinization temperature of the grain, and wherein the concentration of the alpha amylase is between about 5 to about 20 AAU/gds.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 ground or fractionated grain is selected from the group consisting of: corn claim 1 , corn endosperm claim 1 , milo claim 1 , rice claim 1 , and any combination thereof.5. The method of claim 1 , wherein greater than about 90% of the starch from the ground or fractionated ...

LOW TEMPERATURE METHOD FOR MAKING HIGH GLUCOSE SYRUP

The present teachings provide a method for making a high glucose syrup at a low temperature. In some embodiments, the syrup contains reduced reversion reaction products. The method comprises contacting a starch substrate at a temperature below the starch gelatinization temperature with an enzyme blend comprising a high dose of alpha-amylase and a low dose of glucoamylase. In some embodiments, a blend of two glucoamylases is employed. In some embodiments, a debranching enzyme such as a pullulanase is employed. In some embodiments, the enzymes are staged by adding at different times, or at different temperatures. The present teachings provide for high glucose syrups with fewer reversion products, and allow for higher starch solubilization. 1. A method of making a glucose syrup from refined granular starch slurry comprising;contacting the refined granular starch slurry at a temperature at or below the initial starch gelatinization temperature with a dose of at least 8 AAU/gds of an alpha-amylase, and, a dose of 0.05 GAU/gds to no more than 0.3 GAU/gds of glucoamylase, and,making a glucose syrup.2. The method according to wherein the glucose syrup comprises a DP1 of at least 90%.3. The method according to wherein at least 80% of the refined granular starch is solubilized.4. The method according to wherein the glucose syrup comprises a DP2 of less than 3%.5. The method according to wherein the refined granular starch slurry comprises an initial dry solids content (DS) of 31%-44% or 33-37%.6. The method according to wherein the glucoamylase comprises a mixture of glucoamylases claim 1 , the mixture comprising a fast hydrolyzing glucoamylase and a low reversion glucoamylase.7HumicolaA. Niger. The method according to wherein the glucoamylase comprises a mixture of glucoamylases claim 1 , wherein the mixture of glucoamylases comprises a fast hydrolyzing glucoamylase and a low reversion glucoamylase claim 1 , wherein the fast hydrolyzing glucoamylase is glucoamylase and ...

BUTTIAUXELLA SP. PHYTASE VARIANTS

Provided herein are variants of sp. phytases that may be used in industrial applications including methods for starch liquefaction, alcohol fermentations and for enhancing phosphate digestion in foods and animal feeds. 115-. (canceled)16Buttiauxella. A phytase variant having phytase activity and an amino acid sequence that varies from the amino acid sequence of wild type sp. phytase (SEQ ID NO: 1) , wherein the amino acid sequence of the phytase variant comprises at least one variation as compared with SEQ ID NO: 1 , and wherein the phytase sequence variation comprisesa) N37Y, S75P, A89T, D92A, T134I, H160R, F164E, T171V, T176K, A178P, S188P, G192A, K198R, K207E, A209S, S248L, Q256Y, A261E, N270K, A374Pb) N37Y, G77S, A89T, D92A, T134I, H160R, F164E, T171V, T176K, A178P, S188P, G192A, K198R, K207E, A209S, S248L, Q256Y, A261E, N270K, A374Pc) N37Y, S75P, Q76R, A89T, D92A, T134I, H160R, F164E, T171I, T176K, A178P, S188P, G192A, K207E, A209S, A235V, S248L, Q256Y, A261E, N270K, A374Pd) N37Y, A89T, D92A, T134I, F164E, T171V, T176K, A178P, G192A, K207E, A209S, A235V, S248L, Q256P, A261E, N270K, A374Pe) S75P, Q76R, A89T, D92A, T134I, H160R, F164E, T171I, T176K, A178P, S188P, G192A, K207E, A209S, S248L, Q256Y, A261E, N270K, A374Pf) N37Y, Q76R, A89T, D92A, T134I, H160R, F164E, T171I, T176K, A178P, S188P, G192A, K207E, A209S, S248L, Q256Y, A261E, N270K, A374Pg) N37Y, Q76R, A89T, D92A, T134I, F164S, T171V, T176K, A178P, S188P, G192A, K207E, A209S, A235V, S248L, Q256A, A261E, N270K, A374Ph) S75P, A89T, D92A, T134I, F164E, T171V, T176K, A178P, S188P, G192A, K207E, A209S, A235V, S248L, Q256Y, A261E, N270K, A374Pi) S75P, Q76R, A89T, D92A, T134I, H160R, F164E, T171V, T176K, A178P, S188P, G192A, K207E, A209S, A235V, S248L, Q256Y, A261E, N270K, P367L, A374Pj) N37Y, A89T, D92A, T134I, F164E, T171I, T176K, A178P, G192A, K207E, A209S, A235V, S248L, Q256Y, A261E, N270K, A374Pk) N37Y, Q76R, A89T, D92A, T134I, F164E, T171V, T176K, A178P, G192A, K207E, A209S, S248L, Q256Y, A261E, N270K, ...

PROCESSES OF TREATING GRAIN

Processes of treating grain (e.g., corn), involving milling the grain to produce milled grain wherein the grain germ remains intact in the milled grain, and producing a mixture by mixing the milled grain with water and at least one enzyme selected from the group consisting of protease, alpha amylase, glucoamylase, cell wall degrading enzyme, and mixtures thereof, wherein the pH of the mixture is optionally adjusted to a pH of about 3.5 to about 6.5, and incubating the mixture for about 1 to about 3 hours to produce an incubated mixture. 1. A process of treating grain , said process comprising milling said grain to produce milled grain wherein the grain germ remains intact in said milled grain , and producing a mixture by mixing said milled grain with water and protease and optionally at least one enzyme selected from the group consisting of alpha amylase , glucoamylase , cell wall degrading enzyme , and mixtures thereof , wherein the pH of said mixture is optionally adjusted to a pH of about 3.5 to about 6.5 , and incubating said mixture for about 1 to about 3 hours to produce an incubated mixture;said process further comprising:(a) separating said incubated mixture into germ and a liquid slurry, reducing the particle size of materials in said liquid slurry by grinding, adding yeast and amylase to said liquid slurry and fermenting for about 30 to about 72 hours, removing ethanol from said liquid slurry and separating said liquid slurry into fiber and a defibered liquid slurry, and separating said defibered liquid slurry to produce protein solids; or(b) separating said incubated mixture into germ and a liquid slurry, reducing the particle size of materials in said liquid slurry by grinding, adding amylase to said liquid slurry to hydrolyze the starches in said liquid slurry, and separating said liquid slurry into fiber and a defibered liquid slurry, and separating said defibered liquid slurry into protein solids and a solution containing sugars; or(c) separating said ...

TRICHODERMA REESEI HOST CELLS EXPRESSING A GLUCOAMYLASE FROM ASPERGILLUS FUMIGATUS AND METHODS OF USE THEREOF

Fungal glucoamylases from —expressed in host cells (AfGATR) are provided. host cells express AfGATRs at higher, or at least comparable, levels to natively expressed AfGA . AfGATRs, including AfGA1TR and AfGA2TR, exhibit high activity at elevated temperatures and at low pH, so AfGATRs can be used efficiently in a process of saccharification in the presence of alpha-amylase, such as alpha-amylase (AkAA). AfGATRs advantageously catalyze starch saccharification to an oligosaccharide composition significantly enriched in DP1 (i.e., glucose) compared to the products of saccharification catalyzed by glucoamylase (AnGA) or native AfGA expressed in . AfGATRs such as AfGA1TR, AfGA2TR or a variant thereof can be used at a lower dosage than AnGA and natively expressed AfGAs to produce comparable levels of glucose. 1Trichoderma reeseiA. fumigatus. A recombinant host cell expressing an AfGATR , or variant thereof , having at least 80% sequence identity to SEQ ID NO: 12 or 13 , wherein the AfGATR , or variant thereof , is more thermostable than an AfGA , or variant thereof , having the same amino acid sequence of AfGATR , or variant thereof , and wherein the AfGA , or variant thereof , is expressed in an host cell.2. A recombinant AfGATR claim 1 , or variant thereof claim 1 , produced by the host cell of .3. The recombinant AfGATR claim 2 , or variant thereof claim 2 , of claim 2 , wherein said AfGATR claim 2 , or variant thereof claim 2 , has at least 70% activity at 74° C. at pH 5.0 over 10 min.4. The recombinant AfGATR claim 3 , or variant thereof claim 3 , of claim 3 , wherein said AfGATR claim 3 , or variant thereof claim 3 , is an AfGA1TR claim 3 , or variant thereof.5. The recombinant AfGA1TR claim 4 , or variant thereof claim 4 , of claim 4 , wherein said AfGA1TR claim 4 , or variant claim 4 , thereof has at least 70% activity over a temperature range of 55° to 74° C. at pH 5.0 over 10 min.6. The recombinant AfGA1TR claim 5 , or variant thereof claim 5 , of claim 5 , ...

Barley-Based Biorefinery Process

The barley-based biorefinery process comprises a method of optimizing the production of ethanol and value-added products from barley feedstock. Specifically, the biorefinery process is an integrated barley treatment process that utilizes essentially all components of barley (including the barley hulls) to efficiently produce ethanol and other value-added liquids and solids. 1. A method of processing barley to co-produce ethanol and value-added products , the method comprising the steps of:(a) separating barley hulls from barley endosperm;(b) treating the hulls with alpha-amylase and glucoamylase to produce a glucose solution and destarched hulls;(c) treating the destarched hulls to produce pretreated hulls by soaking the destarched hulls in aqueous ammonia, or soaking in ethanol and aqueous ammonia, or by treating with anhydrous ammonia;(d) hydrolyzing the pretreated hulls with hemicellulases to produce a xylo se solution and residual solids;(e) further hydrolyzing the residual solids with cellulases to produce a glucose solution;(f) using the glucose solutions obtained in (b) and (e) as mashing waters to produce ethanol in addition to ethanol produced from starch in the barley endosperm, or optionally to produce a same amount of ethanol from a reduced quantity of barley endosperm; and,(g) using the xylose solution obtained in (d) for production of value-added products.2. The method of wherein step (b) is replaced by a step comprising treatment of starch within the hulls by endogenous beta-amylase contained in the hulls and optionally other suitable debranching enzymes to produce a maltose solution and destarched hulls.3. The method of wherein between steps (b) and (c) the destarched hulls are washed with water or a buffer to produce additional glucose and an additional solid/liquid separation step occurs before the destarched hulls proceed to step (c).4. The method of wherein the step described in is repeated until about half of an expected total amount of glucose ...

PRODUCTION OF ISOPRENE UNDER NEUTRAL pH CONDITIONS

Embodiments of the present disclosure relate to a process for producing isoprene 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. 1Trichoderma reesei. A method for producing isoprene comprising culturing a host cell , which comprises a heterologous nucleic acid encoding an isoprene 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 6.5 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 glucoamylase (TrGA) comprising SEQ ID NO: 6 , or a variant thereof , and wherein the variant has at least 99% sequence identity to the parent glucoamylase , wherein the host cell is selected from the group consisting of bacterial cells and fungal cells.2. The method of claim 1 , wherein the variant has one amino acid modification compared to the parent glucoamylase.34-. (canceled)5. The method of claim 1 , wherein the TrGA is SEQ ID No: 6.6. (canceled)7. The method of claim 1 , wherein SSF are carried out at pH 6.5 to 7.5.8. The method of claim 1 , wherein SSF are carried out at pH 7.0 to 7.5.9. The method of claim 1 , wherein SSF is performed at a temperature in a range of about 30° C. to about 60° C.10. The method of claim 9 , wherein SSF is performed at a temperature in a range of about 40° C. to about 60° C.11. The method of claim 1 , wherein the starch substrate is about 15% to 50% dry solid (DS).12. The method of claim 1 , wherein the starch substrate ...

Methods for Obtaining Corn Oil from Milled Corn Germ

Methods for obtaining corn oil from milled corn germ (e.g., dry milled corn germ), involving adding water, at least one acidic cellulase, at least one acidic protease, and at least one phytase to milled corn germ to obtain corn oil. 1. A method for obtaining corn oil from milled corn germ , comprising adding water , at least one acidic cellulase , at least one acidic protease , and optionally at least one phytase to said milled corn germ to obtain corn oil.2. The method according to claim 1 , Wherein said method is conducted for about 1 to about 40 hours.3. The method according to claim 1 , Wherein said method is conducted at about 25° to about 70° C.4. The method according to claim 1 , wherein said method is conducted at about 35° to about 60° C.5. The method according to claim 1 , wherein said method is conducted at about 40° to about 55° C.6. The method according to claim 1 , wherein said method is conducted at about 50° C.7. The method according to claim 1 , wherein said method is conducted at a pH of about 3 to about 6.8. The method according to claim 1 , wherein said method is conducted at a pH of about 3.5 to about 5.5.9. The method according to claim 1 , wherein said method is conducted at a pH of about 4 to about 5.10. The method according to claim 1 , wherein said method utilizes about 1 to about 5 kg acidic cellulase per Metric Ton (MT) and about 1 to about 5 kg acidic protease per MT milled corn germ and optionally about 0.05 to about 5 kg phytase per MT milled corn germ.11. The method according to claim 1 , wherein said method utilizes at least one phytase.12. The method according to claim 11 , wherein said method utilizes about 1 to about 5 kg acidic cellulase Per Metric Ton (MT) and about 1 to about 5 kg acidic protease per MT milled corn germ and about 0.05 to about 5 kg phytase per MT milled corn germ.13. The method according to claim 1 , wherein said milled corn germ is selected from the group consisting of dry milled corn germ claim 1 , wet milled ...

ENZYMATIC HYDROLYSIS OF DISACCHARIDES AND OLIGOSACCHARIDES USING ALPHA-GLUCOSIDASE ENZYMES

A method is disclosed for hydrolyzing an alpha-1,5 glucosyl-fructose linkage in a saccharide (disaccharide or oligosaccharide) such as leucrose. This method comprises contacting the saccharide with an alpha-glucosidase enzyme such as transglucosidase or glucoamylase under suitable conditions, during which contacting step the enzyme hydrolyzes at least one alpha-1,5 glucosyl-fructose linkage of the saccharide. This method is useful for reducing the amount of leucrose in a filtrate isolated from a glucan synthesis reaction, for example. 1. A method of hydrolyzing an alpha-1 ,5 glucosyl-fructose linkage in a saccharide comprising at least one alpha-1 ,5 glucosyl-fructose linkage , wherein the saccharide is a disaccharide or oligosaccharide , and wherein the method comprises:contacting the saccharide with an alpha-glucosidase enzyme under suitable conditions, wherein said alpha-glucosidase enzyme hydrolyzes at least one alpha-1,5 glucosyl-fructose linkage of the saccharide,and wherein the amount of the saccharide is reduced compared to the amount of the saccharide that was present prior to said contacting.2. The method of claim 1 , wherein the alpha-glucosidase enzyme is immobilized.3. The method of claim 1 , wherein the saccharide is leucrose.4. The method of claim 3 , wherein the concentration of leucrose after the contacting step is less than 50% of the concentration of leucrose that was present prior to said contacting.5. The method of claim 1 , wherein the suitable conditions comprise:(i) a glucan synthesis reaction, or(ii) a fraction obtained from the glucan synthesis reaction;wherein the saccharide is a byproduct of the glucan synthesis reaction.6. The method of claim 5 , wherein the glucan synthesis reaction produces at least one insoluble alpha-glucan product.7. The method of claim 6 , wherein the fraction is a filtrate of the glucan synthesis reaction.8. The method of claim 5 , wherein the glucan synthesis reaction produces at least one soluble alpha-glucan ...

ENZYMATIC HYDROLYSIS OF DISACCHARIDES AND OLIGOSACCHARIDES USING ALPHA-GLUCOSIDASE ENZYMES

A method is disclosed for hydrolyzing an alpha-1,3 or alpha-1,6 glucosyl-glucose linkage in a saccharide (disaccharide or oligosaccharide). This method comprises contacting the saccharide with an alpha-glucosidase enzyme such as transglucosidase under suitable conditions, during which contacting step the enzyme hydrolyzes at least one alpha-1,3 or alpha-1,6 glucosyl-glucose linkage of the saccharide. This method is useful for reducing the amount of oligosaccharides in a filtrate isolated from a glucan synthesis reaction, for example. 1. A method of hydrolyzing an alpha-1 ,3 or alpha-1 ,6 glucosyl-glucose linkage in a saccharide comprising at least one alpha-1 ,3 or alpha-1 ,6 glucosyl-glucose linkage , wherein the saccharide is a disaccharide or oligosaccharide , and wherein the method comprises:contacting the saccharide with an alpha-glucosidase enzyme under suitable conditions, wherein said alpha-glucosidase enzyme hydrolyzes at least one alpha-1,3 or alpha-1,6 glucosyl-glucose linkage of the saccharide,and wherein the amount of the saccharide is reduced compared to the amount of the saccharide that was present prior to said contacting.2. The method of claim 1 , wherein the alpha-glucosidase enzyme is immobilized.3. The method of claim 1 , wherein the degree of polymerization of the saccharide before hydrolysis is 3 to 7.4. The method of claim 1 , wherein the concentration of the saccharide after the contacting step is less than 50% of the concentration of the saccharide that was present prior to said contacting.5. The method of claim 1 , wherein the suitable conditions comprise (i) a glucan synthesis reaction claim 1 , or (ii) a fraction obtained from the glucan synthesis reaction;wherein the saccharide is a byproduct of the glucan synthesis reaction.6. The method of claim 5 , wherein the glucan synthesis reaction produces at least one insoluble alpha-glucan product.7. The method of claim 6 , wherein the fraction is a filtrate of the glucan synthesis reaction.8. ...

Endoglucanase For Reducing The Viscosity Of A Plant Materials Slurry

The present disclosure relates to composition comprising EG cellulase and methods of use, thereof. The compositions are useful, e.g., for reducing the viscosity of plant material slurry. 1. A method for reducing the viscosity of a plant material slurry selected from the group consisting of barley or oats , comprising adding to the slurry a composition comprising an isolated endoglucanase (EG) cellulase , wherein the viscosity in the plant material slurry is primarily due to the presence of betaglucan , and wherein an additional cellulase is separately added to the slurry but is not required to reduce the viscosity of the slurry.2. The method of claim 1 , wherein the EG cellulase is expressed in a filamentous fungus.3Trichoderma reesei.. The method of claim 1 , wherein the EG cellulase is expressed in4. The method of claim 1 , wherein the EG cellulase is expressed under control of the cbh1 promoter.5. The method of claim 1 , wherein the EG cellulase is purified to at least 70% of total protein in the composition.6. The method of claim 1 , wherein the EG cellulase is purified to at least 80% of total protein in the composition.7. The method of claim 1 , wherein the EG cellulase is purified to at least 90% of total protein in the composition.8. The method of claim 1 , wherein the EG cellulase is purified to at least 95% of total protein in the composition.9. The method of claim 1 , wherein the EG cellulase is purified to at least 97% of total protein in the composition.10Trichoderma reeseiHypocrea jecorina. The method of claim 1 , wherein the EG cellulase is a () EG cellulase.11. The method of claim 1 , wherein the reduction in viscosity as a result of adding the EG cellulase is at least equivalent to a reduction in viscosity as a result of adding a mixture of cellulases wherein EG is a component.12. The method of claim 1 , wherein the EG cellulase is selected from the group consisting of EG I claim 1 , EG II claim 1 , and EG III.13. The method of claim 1 , wherein the ...

Polypeptides having glucoamylase activity and method of producing the same

The present invention relates to polypeptides having glucoamylase activity with improved properties and to compositions comprising these polypeptides suitable for use in the production of a food, beverage (e.g. beer), feed, or biofuel. Also described is an improved and cost-effective process for isolating glucoamylases suitable for large scale protein purification procedures. Furthermore, different methods and uses related to glucoamylases according to the invention are disclosed, such as in a brewing process.

EXPRESSION OF GRANULAR STARCH HYDROLYZING ENZYMES IN TRICHODERMA AND PROCESS FOR PRODUCING GLUCOSE FROM GRANULAR STARCH SUSTRATES

The present invention relates to filamentous fungal host cells and particularly host cells useful for the production of heterologous granular starch hydrolyzing enzymes having glucoamylase activity (GSHE). Further the invention relates to a method for producing a glucose syrup comprising contacting a granular starch slurry obtained from a granular starch substrate simultaneously with an alpha amylase and a GSHE at a temperature equal to or below the gelatinization temperature of the granular starch to obtain a composition of a glucose syrup. 118-. (canceled)19. A process for producing residual starch from a granular starch slurry , said process comprising:contacting a granular starch slurry obtained from a granular starch substrate simultaneously with an alpha amylase and a glucoamylase, wherein the glucoamylase has at least 97% amino acid sequence identity with SEQ ID NO: 3, at a temperature equal to or below the gelatinization temperature of the granular starch;allowing the alpha amylase and the glucoamylase to act for a period of time sufficient to hydrolyze the granular starch to obtain glucose and residual starch; and,retaining the residual starch to produce residual starch from the granular starch slurry.20. The method of wherein the residual starch is used for the production of end products.21. The method of wherein the residual starch is recycled and contacted with the alpha amylase and glucoamylase at the temperature equal to or below the gelatinization temperature of the granular starch. The present application is a continuation U.S. application Ser. No. 13/664,216, filed Oct. 30, 2012, which is a continuation of U.S. application Ser. No. 13/230,707, filed Sep. 12, 2011, now U.S. Pat. No. 8,323,932, which is a continuation of U.S. application Ser. No. 11/875,279, filed Oct. 19, 2007, now U.S. Pat. No. 8,034,590, which is a divisional application of U.S. patent application Ser. No. 10/991,654, filed Nov. 18, 2004, now U.S. Pat. No. 7,303,899, and claims ...

Enzymatic hydrolysis of disaccharides and oligosaccharides using alpha-glucosidase enzymes

A method is disclosed for hydrolyzing an alpha-1,5 glucosyl-fructose linkage in a saccharide (disaccharide or oligosaccharide) such as leucrose. This method comprises contacting the saccharide with an alpha-glucosidase enzyme such as transglucosidase or glucoamylase under suitable conditions, during which contacting step the enzyme hydrolyzes at least one alpha-1,5 glucosyl-fructose linkage of the saccharide. This method is useful for reducing the amount of leucrose in a filtrate isolated from a glucan synthesis reaction, for example.

ENZYMATIC HYDROLYSIS OF DISACCHARIDES AND OLIGOSACCHARIDES USING ALPHA-GLUCOSIDASE ENZYMES

A method is disclosed for hydrolyzing an alpha-1,3 or alpha-1,6 glucosyl-glucose linkage in a saccharide (disaccharide or oligosaccharide). This method comprises contacting the saccharide with an alpha-glucosidase enzyme such as transglucosidase under suitable conditions, during which contacting step the enzyme hydrolyzes at least one alpha-1,3 or alpha-1,6 glucosyl-glucose linkage of the saccharide. This method is useful for reducing the amount of oligosaccharides in a filtrate isolated from a glucan synthesis reaction, for example. 1. A method of hydrolyzing an alpha-1 ,3 or alpha-1 ,6 glucosyl-glucose linkage in a saccharide comprising at least one alpha-1 ,3 or alpha-1 ,6 glucosyl-glucose linkage , wherein the saccharide is a disaccharide or oligosaccharide , and wherein the method comprises:contacting the saccharide with an alpha-glucosidase enzyme under suitable conditions, wherein said alpha-glucosidase enzyme hydrolyzes at least one alpha-1,3 or alpha-1,6 glucosyl-glucose linkage of the saccharide,and wherein the amount of the saccharide is reduced compared to the amount of the saccharide that was present prior to said contacting.2. The method of claim 1 , wherein the alpha-glucosidase enzyme is immobilized.3. The method of claim 1 , wherein the degree of polymerization of the saccharide before hydrolysis is 3 to 7.4. The method of claim 1 , wherein the concentration of the saccharide after the contacting step is less than 50% of the concentration of the saccharide that was present prior to said contacting.5. The method of claim 1 , wherein the suitable conditions comprise (i) a glucan synthesis reaction claim 1 , or (ii) a fraction obtained from the glucan synthesis reaction;wherein the saccharide is a byproduct of the glucan synthesis reaction.6. The method of claim 5 , wherein the glucan synthesis reaction produces at least one insoluble alpha-glucan product.7. The method of claim 6 , wherein the fraction is a filtrate of the glucan synthesis reaction.8. ...

Processes of Treating Grain

Processes of treating grain (e.g., corn), involving milling the grain to produce milled grain wherein the grain germ remains intact in the milled grain, and producing a mixture by mixing the milled grain with water and at least one enzyme selected from the group consisting of protease, alpha amylase, glucoamylase, cell wall degrading enzyme, and mixtures thereof, wherein the pH of the mixture is optionally adjusted to a pH of about 3.5 to about 6.5, and incubating the mixture for about 1 to about 3 hours to produce an incubated mixture. 1. A process of treating grain , said process comprising milling said grain to produce milled grain wherein the grain germ remains intact in said milled grain , and producing a mixture by mixing said milled grain with water and at least one enzyme selected from the group consisting of protease , alpha amylase , glucoamylase , cell wall degrading enzyme , and mixtures thereof , wherein the pH of said mixture is optionally adjusted to a pH of about 3.5 to about 6.5 , and incubating said mixture for about 1 to about 3 hours to produce an incubated mixture.2. The process of claim 1 , further comprising separating said incubated mixture into germ and a liquid slurry claim 1 , reducing the particle size of materials in said liquid slurry by grinding claim 1 , adding yeast and amylase to said liquid slurry and fermenting for about 30 to about 72 hours claim 1 , removing ethanol from said liquid slurry and separating said liquid slurry into fiber and a defibered liquid slurry claim 1 , and separating said defibered liquid slurry to produce protein solids.3. The process of claim 1 , further comprising separating said incubated mixture into germ and a liquid slurry claim 1 , reducing the particle size of materials in said liquid slurry by grinding claim 1 , adding amylase to said liquid slurry to hydrolyze the starches in said liquid slurry claim 1 , and separating said liquid slurry into fiber and a defibered liquid slurry claim 1 , and ...

Alpha-amylase blends and methods for using said blends

The present invention relates to an alpha-amylase blend, including a B. stearothermophilus alpha-amylase (AmyS) wherein the amino acid at position S242 is substituted and a B. licheniformis alpha-amylase The invention also relates to processes using the alpha-amylase blends for starch liquefaction and saccharification, ethanol production, and a sweetener production.

Acid fungal protease in fermentation of insoluble starch substrates

The invention is directed to methods of producing ethanol and decreasing residual starch production in a no cook fermentation comprising contacting granular starch containing substrates with a granular starch hydrolyzing enzyme, a protease, and a fermenting microorganism under suitable fermentation conditions at a temperature below the starch gelatinization temperature of the starch substrate to produce ethanol, wherein the ethanol production is increased and the amount of residual starch is decreased compared to a substantially similar method conducted without the protease.

Process for conversion of granular starch to ethanol

A method is disclosed for producing glucose from a granular starch substrate including, contacting a slurry comprising granular starch obtained from plant material with an alpha-amylase at a temperature below the starch gelatinization temperature of the granular starch to produce oligosaccharides and hydrolyzing the oligosaccharides to produce a mash comprising at least 20% glucose and further comprising fermenting the mash to obtain ethanol.

Production of ethanol from barley and DDGS containing reduced beta-glucan and phytic acid

Described herein is a method of preparing DDGS containing reduced levels of beta-glucan and phytic acid suitable for an animal feed.

Trichoderma reesei host cells expressing a glucoamylase from aspergillus fumigatus and methods of use thereof

Fungal glucoamylases from Aspergillus fumigatus - expressed in Trichoderma reesei host cells (AfGATR) are provided. Trichoderma reesei host cells express AfGATRs at higher, or at least comparable, levels to natively expressed AfGA Aspergillus fumigatus . AfGATRs, including AfGA1TR and AfGA2TR, exhibit high activity at elevated temperatures and at low pH, so AfGATRs can be used efficiently in a process of saccharification in the presence of alpha-amylase, such as Aspergillus kawachii alpha-amylase (AkAA). AfGATRs advantageously catalyze starch saccharification to an oligosaccharide composition significantly enriched in DP1 (i.e., glucose) compared to the products of saccharification catalyzed by Aspergillus niger glucoamylase (AnGA) or native AfGA expressed in Aspergillus fumigatus . AfGATRs such as AfGA1TR, AfGA2TR or a variant thereof can be used at a lower dosage than AnGA and natively expressed AfGAs to produce comparable levels of glucose.

Acid fungal protease in fermentation of insoluble starch substrates

The invention is directed to methods of producing ethanol and decreasing residual starch production in a no cook fermentation comprising contacting granular starch containing substrates with a granular starch hydrolyzing enzyme, a protease, and a fermenting microorganism under suitable fermentation conditions at a temperature below the starch gelatinization temperature of the starch substrate to produce ethanol, wherein the ethanol production is increased and the amount of residual starch is decreased compared to a substantially similar method conducted without the protease.

Process for conversion of granular starch to ethanol

The present invention concerns a method of producing glucose from a granular starch substrate comprising, contacting a slurry comprising granular starch obtained from plant material with an alpha-amylase at a temperature below the starch gelatinization temperature of the granular starch to produce oligosaccharides and hydrolyzing the oligosaccharides to produce a mash comprising at least 20% glucose and further comprising fermenting the mash to obtain ethanol.

Alpha-amylase blend for starch processing and method of use thereof

The present disclosure relates to an enzyme blend comprising a low pH, thermostable alpha-amylase and a Bacillus licheniformis alpha-amylase. The blend can include at least about 1.0 Liquefon Unit (LU) of the B. licheniformis alpha-amylase for every 5.0 Modified Wohlgemuth Unit (MWU) of the low pH, thermostable alpha-amylase. The enzyme blend described is suitable for starch liquefaction and saccharification, ethanol production, and/or sweetener production, among other things. Also provided herein is a method of processing a starch by liquefying the starch with the low pH, thermostable alpha-amylase and the Bacillus licheniformis alpha-amylase, simultaneously or sequentially.

Alpha-amylase blend for starch processing and method of use thereof

The present disclosure relates to an enzyme blend comprising a low pH, thermostable alpha-amylase and a Bacillus licheniformis alpha-amylase. The blend can include at least about 1.0 Liquefon Unit (LU) of the B. licheniformis alpha-amylase for every 5.0 Modified Wohlgemuth Unit (MWU) of the low pH, thermostable alpha-amylase. The enzyme blend described is suitable for starch liquefaction and saccharification, ethanol production, and/or sweetener production, among other things. Also provided herein is a method of processing a starch by liquefying the starch with the low pH, thermostable alpha-amylase and the Bacillus licheniformis alpha-amylase, simultaneously or sequentially.

Yeast host cells epxressing a glucoamylase from aspergillus fumigatus and methods of use thereof

Fungal glucoamylases from Aspergillus fumigatus - expressed in Trichoderma reesei host cells (AfGATR) are provided. Trichoderma reesei host cells express AfGATRs at higher, or at least comparable, levels to natively expressed AfGA Aspergillus fumigatus, AfGATRs, including AfGAlTR and AfGA2TR, exhibit high activity at elevated temperatures and at low pH, so AfGATRs can be used efficiently in a process of saccharification in the presence of alpha-amylase (AkAA), AfGATRs advantageously catalyze starch saccharification to an oligosaccharide composition significantly enriched in DP1 (i.e. glucose) compared to the products of saccharification catalyzed by Aspergillus niger glucoamylase (AnGA) or native AfGA expressed in Aspergillus fumigatus. AfGATRs such as AfGA1TR, AfGA2TR or a variant thereof can be used at a lower dosage than AnGA and natively expressed AfGAs to produce comparable levels of glucose.

Enzyme compositions comprising a glucoamylase, an acid stable alpha amylase, and an acid fungal protease

The present invention relates to an enzyme blend composition comprising a glucoamylase, an acid stable alpha amylase, and an acid fungal protease. The present invention is further directed to a method for producing end products such as alcohols from fermentable sugars, comprising the steps of: (a) contacting a slurry comprising a milled grain that contains starch with an alpha amylase to produce a liquefact; (b) contacting the liquefact with a glucoamylase, an acid stable alpha amylase, and an acid fungal protease, to produce fermentable sugars; and (c) fermenting the fermentable sugars in the presence of a fermenting organism to produce end products.

Methods for producing ethanol from carbon substrates

The present invention provides means for the production of desired end-products of in vitro and/or in vivo bioconversion of biomass-based feed stock substrates, including but not limited to such materials as starch and cellulose. In particularly preferred embodiments, the methods of the present invention do not require gelatinization and/or liquefaction of the substrate. In particularly preferred embodiments, the present invention provides means for the production of ethanol.

Glucoamylase and Buttiauxiella phytase during saccharification

Described are compositions and methods relating to the use of a glucoamylase in combination with a phytase in starch processing to reduce the levels of phytic acid in end-products.

A food product comprising a low temperature rice protein concentrate

Rice protein concentrates prepared at low temperature exhibit improved functionality and beneficial physiological benefits, including lowered cholesterol and enhanced lactic acid dehydrogenase activity, without an increase in blood urea nitrogen. The rice protein concentration could be made into a wet dough with comparatively less water than a soy protein concentrate. Use of the rice protein concentrate thus improved processing steps in the formulation of a food article containing the concentrate. The food product advantageously shows an extending shelf life and improved palatable texture.

Process for alcohol and co-product production from grain sorghum

Described herein are methods for producing alcohol and particularly ethanol from milled sorghum.

Modified forms of pullulanase

The present invention relates to modified pullulanases useful in the starch industry. The present invention provides methods for producing the modified pullulanase, enzymatic compositions comprising the modified pullulanase, and methods for the saccharification of starch comprising the use of the enzymatic compositions.

Variants of bacillus licheniformis alpha-amylase with increased thermostability and/or decreased calcium dependence.

Las variantes de alfa-amilasa de B. licheniformis exhiben un rendimiento enzimático mejorado, que incluyen una termoestabilidad aumentada y dependencia de calcio reducida. Las composiciones comprenden las variantes que son útiles en métodos para el procesamiento del almidón, licuefacción del almidón, fermentación, sacarificación del almidón, limpieza, lavandería, desencolado textil, horneado y eliminación de biopelícula. Los ácidos nucleicos que codifican las variantes también se describen.

Levane sucrase enzyme, process for its preparation, microorganisms which produce it and compositions containing it.

Enzyme stabilization

Methods for stabilizing enzymes in liquid compositions, including those liquid compositions having a high water content and those stabilized enzyme liquid compositions formed thereby. The method involves forming the enzyme, so that it is in an insoluble form thereof and then adding thereto an agent for maintaining the enzyme in the insoluble form thereof. Examples of such insoluble forms are crystal forms of the enzyme. Enzymes which are stabilized in this manner are useful for combining with liquid compositions, including liquid compositions having a high water content. The method is particularly useful for the preparation of stable enzyme-containing liquid detergent compositions.

Production of ethanol from barley and ddgs containing reduced beta-glucan and phytic acid

Described herein is a method of preparing DDGS containing reduced levels of beta-glucan and phytic acid suitable for an animal feed.

Enzyme blends for fermentation

The present invention relates to an enzyme blend composition comprising a glucoamylase, an acid stable alpha amylase, and an acid fungal protease. The present invention is further directed to a method for producing end products such as alcohols from fermentable sugars, comprising the steps of: (a) contacting a slurry comprising a milled grain that contains starch with an alpha amylase to produce a liquefact; (b) contacting the liquefact with a glucoamylase, an acid stable alpha amylase, and an acid fungal protease, to produce fermentable sugars; and (c) fermenting the fermentable sugars in the presence of a fermenting organism to produce end products.

Direct starch to fermentable sugar

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 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.

Production of maltotetraose syrup using a pseudomonas saccharophila maltotetraohydrolase variant

Variants of a Pseudomonas saccharophila G4-forming amylase (PS4) advantageously can catalyze high temperature saccharification to produce maltotetraose syrup from a starch liquefact or granular starch, e.g. , derived from cornstarch. The PS4 variants are useful in a process of saccharification of starch that advantageously produces significant amounts of maltotetraose, which can be used downstream in a process of producing a maltotetraose syrup. In one embodiment, a thermostable PS4 variant is provided that can produce about 40% to about 60% by weight maltotetraose, based on total saccharide content.

Single ph process for starch liquefaction and saccharification for high-density glucose syrups

Embodiments of the present disclosure relate to a process for producing downstream products, such as high-density glucose syrups and high-glucose fermentation feedstock, from a starch-containing substrate without a pH adjustment before the saccharification step. The saccharification is effectively catalyzed by a glucoamylase at a pH in the range of 5.2 to 5.6.

Dry solids staging fermentation process

A dry solids staging fermentation process for producing an end-product, such as ethanol is disclosed said process including an initial fermentation step including combining a first fermentable substrate with one or more starch hydrolyzing enzymes and fermenting organisms in a fermentation vessel and a loading step which includes adding a second fermentable substrate to the fermentation vessel wherein the percent dry solids (% DS) of the fermentation broth increases over time.

Alpha-amylase variants with altered properties

Disclosed are compositions comprising variants of alpha-amylase that have alpha-amylase activity and which exhibit altered properties relative to a parent AmyS-like alpha-amylase from which they are derived. The compositions comprise an additional enzyme such as a phytase. Also disclosed are methods of using the compositions, and kits related thereto.

Expression of granular starch hydrolyzing enzymes in trichoderma and process for producing glucose from granular starch substrates

Modified forms of pullulanase

A truncated Bacillus pullulanase comprising a deletion of about 100 amino acids from the amino terminus of a pullulanase obtainable from Bacillus deramificans, wherein said truncated pullulanase comprises a conserved Y region, and is capable of catalyzing the hydrolysis of an alpha-1,6-glucosidic bond. (62) Divided out of 506290

Barley-based biorefinery process

The barley-based biorefinery process comprises a method of optimizing the production of ethanol and value-added products from barley feedstock. Specifically, the biorefinery process is an integrated barley treatment process that utilizes essentially all components of barley (including the barley hulls) to efficiently produce ethanol and other value-added liquids and solids.

Acid fungal protease in fermentation of insoluble starch substrates

The invention is directed to methods of producing ethanol and decreasing residual starch production in a no cook fermentation comprising contacting granular starch containing substrates with a granular starch hydrolyzing enzyme, a protease, and a fermenting microorganism under suitable fermentation conditions at a temperature below the starch gelatinization temperature of the starch substrate to produce ethanol, wherein the ethanol production is increased and the amount of residual starch is decreased compared to a substantially similar method conducted without the protease.

Alpha-amylase variants with altered properties

Disclosed are compositions comprising variants of alpha-amylase that have alpha-amylase activity and which exhibit altered properties relative to a parent AmyS-like alpha-amylase from which they are derived. The compositions comprise an additional enzyme such as a phytase. Also disclosed are methods of using the compositions, and kits related thereto.

alpha-amylase variants with altered properties.

Anhydride modified microbial protein having reduced nucleic acid levels

The process of the invention comprises disrupting microbial protein containing cells; derivatizing the resultant mixture comprising protein and nucleic acid with an acid anhydride; and then isoelectrically precipitating a nucleic acid diminished protein containing fraction from a nucleic acid enriched supernatant.

Aspergillus Kawachi Acid-Stable Alpha Amylase and Applications in Granular Starch Hydrolysis

The present invention relates to a method for producing an end product and particularly an alcohol which comprises contacting a granular starch substrate with an acid-stable alpha amylase (asAA) having granular starch hydrolyzing (GSH) activity and a glucoamylase (GA) in a fermentation step which also comprises ethanologenic microorganisms at a temperature between about 25 to 65° C. and producing an end-product.

Acid-stable alpha amylases having granular starch hydrolyzing activity and enzyme compositions

The present invention relates to an acid-stable alpha amylase (asAA) derived from a strain of Aspergillus kawachi, which has granular starch hydrolyzing (GSH) activity, the heterologous expression of the asAA having GSH activity in filamentous fungal host cells and enzyme compositions including the same which optionally include glucoamylase.

Acid fungal protease in fermentation of insoluble starch substrates

The invention is directed to methods of producing ethanol and decreasing residual starch production in a no cook fermentation comprising contacting granular starch containing substrates with a granular starch hydrolyzing enzyme, a protease, and a fermenting microorganism under suitable fermentation conditions at a temperature below the starch gelatinization temperature of the starch substrate to produce ethanol, wherein the ethanol production is increased and the amount of residual starch is decreased compared to a substantially similar method conducted without the protease.

Methods for producing end-products from carbon substrates

The present invention provides means for the production of desired end-products of in vitro and/or in vivo bioconversion of biomass-based feed stock substrates, including but not limited to such materials as starch and cellulose. In particularly preferred embodiments, the methods of the present invention do not require gelatinization and/or liquefaction of the substrate.

Modified forms of pullulanase

The present invention relates to modified pullulanases useful in the starch industry. The present invention provides methods for producing the modified pullulanase, enzymatic compositions comprising the modified pullulanase, and methods for the saccharification of starch comprising the use of the enzymatic compositions.

Liquid lipase from animal origin and method of preparation

Production of isoprene under neutral ph conditions

Embodiments of the present disclosure relate to a process for producing isoprene 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 8Ø 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.

Baking process and a method thereof

A method for improving dough characteristics without added sugar is disclosed. The process comprises of treating cereal flour with an enzyme to produce a composition containing fermentable sugars (glucose, fructose or maltose) and/or isomaltooligosaccharides. The composition can be used to form a dough mixture: shaping the dough mixture and baking the said shaped dough mixture to form baked products.

Methods for obtaining oil from maize using acid protease and cell-wall polysaccharide-degrading enzymes

Disclosed are methods for obtaining oil from maize, involving grinding maize kernels to form flour, adding water to the flour to form a slurry, and incubating the slurry with ?-amylase for about 10 minutes to about 180 minutes at a temperature of about 75° to about 120°C and at a pH of about 3 to about 7 to form a mash, cooling the mash to about 15°C to about 40°C and adding a nitrogen source, glucoamylase, yeast, acid protease, and cell-wall polysaccharide-degrading enzymes to form a beer containing ethanol and oil, wherein the beer has a pH of about 3 to about 7, and recovering oil from the beer.

Expression of granular starch hydrolyzing enzymes in trichoderma and process for producing glucose from granular starch substrates

The present invention relates to filamentous fungal host cells and particularly Trichoderma host cells useful for the production of heterologous granular starch hydrolyzing enzymes having glucoamylase activity (GSHE). Further the invention relates to a method for producing a glucose syrup comprising contacting a granular starch slurry obtained from a granular starch substrate simultaneously with an alpha amylase and a GSHE at a temperature equal to or below the gelatinization temperature of the granular starch to obtain a composition of a glucose syrup.

Recovery of microbial lipase

Disclosed is a method for causing the dissociation of microbial mycelium and extracellular lipase bound thereto and increasing the measurable activity of the lipase. The method involves treating an aqueous suspension of the mycelium with the anhydride of a dicarboxylic acid which results in dissociation of the mycelium and lipase thereby facilitating recovery of the lipase.

Improved fermentation using molasses

Extracting lipase from mammalian tissue

Recovery of microbial lipase

Abstract of the Disclosure Disclosed is a method for causing the dissocia-tion of microbial mycelium and extracellular lipase bound thereto and increasing the measurable activity of the lipase. The method involves treating an aqueous suspension of the mycelium with the anhydride of a dicarboxylic acid which results in dissociation of the mycelium and lipase thereby facilitating recovery of the lipase.

Heterologous expression of an aspergillus kawachi acid-stable alpha amylase and applications in granular starch hydrolysis

The present invention relates to a granular starch hydrolyzing enzyme composition comprising an acid stable alpha amylase (asAA) having granular starch hydrolyzing activity. The invention also relates to a one-step method for producing an alcohol which comprises contacting a granular starch substrate with an acid-stable alpha amylase (asAA) having granular starch hydrolyzing (GSH) activity and a glucoamylase (GA) in a fermentation step which also comprises ethanologenic microorganisms at a temperature of 25 -40~C to obtain a fermentation broth having 5 to 20% ethanol.

A process for hydrolyzing starch without pH adjustment

An alpha-amylase enzyme obtained from Bacillus acidocaldarius species is utilized to liquefy starch at a pH as low as 3.0 without the need to add thermostabilizing agents such as calcium. The alpha-amylase produces acceptable DE yields in a single liquefaction step and does not need to be inactivated prior to conducting saccharification which can proceed without adjustment of the pH of the liquefact. Alternatively, a secondary liquefaction process can be utilized, in which case two additions of the alpha-amylase are used resulting in a combined low dosage of the enzyme.

Ph adjustment free system for producing fermentable sugars and alcohol

The present invention relates to a process for producing downstream products, such as fermentable sugars (e.g., glucose) and alcohols (e.g., ethanol) from starch-containing material (e.g., grain) without a pH adjustment before or after the starch liquefaction step.

Methods for producing end-products from carbon substrates

The present invention provides means for the production of desired end-products of in vitro and/or in vivo bioconversion of biomass-based feed stock substrates, including but not limited to such materials as starch and cellulose. In particularly preferred embodiments, the methods of the present invention do not require gelatinization and/or liquefaction of the substrate.

Liquid lipase from animal origin and method of preparation

Disclosed is a method for the extraction of lipase from mammalian tissue with which it is associated which involves contacting the tissue with an alkaline, aqueous medium having a pH of from greater than 7.0 up to a level at which the alkalinity will deactivate the lipase.

Expression of granular starch hydrolyzing enzymes in Trichoderma and process for producing glucose from granular starch substrates

The present invention relates to filamentous fungal host cells and particularly Trichoderma host cells useful for the production of heterologous granular starch hydrolyzing enzymes having glucoamylase activity (GSHE). Further the invention relates to a method for producing a glucose syrup comprising contacting a granular starch slurry obtained from a granular starch substrate simultaneously with an alpha amylase and a GSHE at a temperature equal to or below the gelatinization temperature of the granular starch to obtain a composition of a glucose syrup.

Dry solids staging fermentation process

A dry solids staging fermentation process for producing an end-product, such as ethanol is disclosed said process including an initial fermentation step including combining a first fermentable substrate with one or more starch hydrolyzing enzymes and fermenting organisms in a fermentation vessel and a loading step which includes adding a second fermentable substrate to the fermentation vessel wherein the percent dry solids (% DS) of the fermentation broth increases over time.