Abstract: Provided are modified beta-glucosidase enzymes, derived from the Trichoderma reesei Cel3A beta-glucosidase, that exhibit improved stability at low pH, low pH and high aeration, low pH and high agitation, or low pH and elevated temperature. Also provided are genetic constructs comprising nucleotide sequences encoding for modified beta-glucosidase enzymes, methods for the production of modified beta-glucosidase enzymes from host strains and the use of the modified beta-glucosidase enzymes in the hydrolysis of cellulose.

Abstract: Provided is a process for the enzymatic hydrolysis of cellulose to produce glucose from a pretreated cellulosic feedstock. The process comprises providing an aqueous slurry of the pretreated cellulosic feedstock that has a water content that is less than about 140% of the maximum water holding capacity of the pretreated cellulosic feedstock. The aqueous slurry of the pretreated cellulosic feedstock is fed to one or more unmixed hydrolysis reactors and hydrolyzed with cellulase enzymes therein. In the unmixed hydrolysis reactor(s), the cellulase enzymes hydrolyze a portion of the cellulose to produce soluble sugars, thereby producing a mixture of partially hydrolyzed cellulose containing soluble sugars. The hydrolysis of the cellulose to glucose is continued by feeding the mixture of partially hydrolyzed cellulose to one or more mixed hydrolysis reactors. Also provided are systems for carrying out the foregoing enzymatic hydrolysis.

Abstract: A modified Trichoderma reesei Family 6 (TrCel6A) cellulase enzyme comprising amino acid substitutions at one or more positions selected from the group consisting of 129, 322, 363 and 410 of SEQ ID NO: 1 is provided. Genetic constructs and genetically modified microbes comprising nucleic sequences encoding the modified TrCel6a cellulase are also provided. The modified TrCel6A cellulase of the invention display at least a 15% decrease in inactivation by lignin relative to a parental TrCel6A cellulase from which the modified TrCel6A is derived. Such cellulases find use in a variety of applications in industry requiring enzymatic hydrolysis of cellulose in the presence of lignin, e.g., the hydrolysis of pretreated lignocellulosic feedstocks for the production of fermentable sugars, sugar alcohols and fuel alcohols.

Abstract: A system for collecting ligno-cellulosic biomass over a large area to enable the commercial refining of biomass from 2,500 to in excess of 50,000 tons of biomass per day to produce ethanol or other products. The biomass is collected at a series of collection points and then transported through a network of conduit “loops” interconnecting each of the collection points and the central refining plant. The water used to transport the biomass, as a slurry, is recovered and sequentially recycled in the same pipeline system to push the biomass slurry around the system in a “loop”.

Abstract: A modified Family 6 cellulase enzyme comprising a proline residue at position 413 is provided. Genetic constructs and genetically modified microbes comprising DNA sequences encoding the modified Family 6 cellulase are also provided. Family 6 cellulases of the invention display improved thermostability, thermophilicity, alkalophilicity, or a combination thereof, relative to the parent Family 6 cellulases. Such cellulases find use in a variety of applications in industry that require cellulase stability and activities at temperatures, pH values, or both, above that of the native enzyme.

Abstract: A fermentation process using hemicellulose-derived carbohydrates for the production of cellulase mixtures with a high proportion of cellulases relative to hemicellulases is provided. The cellulases produced by the process of the invention are further characterized by high specific productivity. The resulting cellulase mixtures comprise at least two times more cellulase than hemicellulase and are useful for the hydrolysis of cellulosic substrated, particularly, pretreated lignocellulosic substrate.

Abstract: A system for collecting ligno-cellulosic biomass over a large area to enable the commercial refining of biomass from 2,500 to in excess of 50,000 tons of biomass per day to produce ethanol or other products. The biomass is collected at a series of collection points and then transported through a network of conduit “loops” interconnecting each of the collection points and the central refining plant. The water used to transport the biomass, as a slurry, is recovered and sequentially recycled in the same pipeline system to push the biomass slurry around the system in a “loop”.

Abstract: Provided is an enzyme mixture for hydrolyzing a pretreated lignocellulosic feedstock to soluble sugars. The enzyme mixture comprises EG4 at a fractional concentration (fEG4) of about 0.25 to about 0.83 (w/w), Swollenin at a fractional concentration (fSwo1) of about 0 to about 0.66 (w/w), and Cip1 at a fractional concentration (fCip1) of 0 to about 0.33 measured relative to all accessory enzymes present in the enzyme mixture. Also provided are processes for converting a pretreated lignocellulosic feedstock to soluble sugars using the enzyme mixtures, and methods of using and producing such enzyme mixtures.

Abstract: The present invention is directed to a continuous process for pretreating a lignocellulosic feedstock. A feedstock slurry is provided, which has a solids concentration of 10% to 33% by weight. The feedstock slurry is pumped through a heating train of at least two stages, each stage including a pump to increase stage pressure and a direct steam injection to heat the feedstock slurry. Acid is also added to the feedstock slurry prior to the heating train, during a heating stage, or after leaving the heating train; the acid is added at a concentration of 0% to 12% weight of acid on weight of initial feedstock. A heated, acidified feedstock slurry is thus produced. The heated, acidified feedstock slurry is flowed through a pre-treatment reactor at a temperature of 160° C. to 280° C. for a time sufficient to increase efficiency of conversion of cellulose in the feedstock to glucose using cellulase enzymes.

Abstract: An enzyme mixture for the enzymatic hydrolysis of a pretreated lignocellulosic feedstock to soluble sugars is provided. Also provided is a process for hydrolyzing a pretreated lignocellulosic feedstock with an enzyme mixture. The enzymatic hydrolysis comprises adding a cellulase enzyme mixture to a pretreated lignocellulosic feedstock. The cellulase enzyme mixture comprises a primary cellulase mixture of CBH1 and CBH2, and EG1 and EG2. The CBH1 and CBH2 being present at greater than or equal to 55% and less than 85% of the primary cellulase mixture. Furthermore, CBH2 is present at a fraction relative to CBH1 and CBH2, and EG2 is present at a fraction relative to the EG1 and EG2.

Abstract: The present invention pertains to a method of converting cellulose to glucose by treating a pretreated lignocellulosic substrate with an enzyme mixture comprising cellulase enzyme and endoglucanase core proteins, wherein the endoglucanase core proteins are present in the enzyme mixture at an amount relative to all endoglucanases from about 35 wt. % to about 100 wt. % and wherein the endoglucanase cellulase enzymes are present in the enzyme mixture at an amount relative to the amount of CBH and EG enzymes from about 2 wt. % to about 50 wt. %. The pretreated lignocellulosic substrate is selected from the group consisting of agricultural residues, residues after starch or sugar removal, dedicated ethanol crops, forestry products, and pulp and paper products, or combinations thereof.

Abstract: A Family 6 cellulase variant enzyme comprising one or more than one amino acid substitution selected from a basic, polar or non-polar amino acid at position 103, a valine or isoleucine at position 136, a tyrosine at position 186, a glutamic acid or glutamine at position 365 and a glutamine at position 410 is provided (said position determined form alignment of the parental Family 6 with SEQ ID NO: 1). Genetic constructs and genetically modified microbes comprising DNA sequences encoding the Family 6 cellulase variant are also provided. Family 6 cellulases of the invention display reduced inhibition by glucose relative to the parent Family 6 cellulases. Such cellulases find use in a variety of applications in industry, e.g., in the hydrolysis of pretreated lignocellulosic feedstock, that require cellulose activity in the presence glucose concentrations that would otherwise inhibit the activity of the parental enzyme.

Abstract: The present invention relates to a method for producing a fermentation product from a sugar hydrolysate. The method comprises fermenting the sugar hydrolysate in a fermentation system with yeast to produce a fermentation broth comprising a fermentation product; introducing acid and an oxidant, such as chlorine dioxide, to the fermentation system so as to expose microbial contaminants in the fermentation system at one or more stages to chlorine dioxide and a pH of less than 3.0; and recovering the fermentation product. In one example of the invention, a yeast slurry obtained from a yeast recycle step is treated with acid and the oxidant.

Abstract: The invention relates to a process for obtaining a product sugar stream from a cellulosic biomass, comprising pretreating the cellulosic biomass to hydrolyze a portion of the cellulose and hemicellulose to produce glucose, acetic acid and a sugar monomer, adding base to the pretreated cellulosic biomass to produce inorganic salt and acetate salt; hydrolyzing the neutralized cellulosic biomass with cellulase enzymes and separating insoluble residue from the resulting sugar stream; treating the clarified sugar stream using exclusion chromatography at a pH of 5 to 10 to produce a product sugar stream.

Abstract: The present invention provides a method of producing xylose from lignocellulosic feedstock. The method comprises disrupting lignocellulosic feedstock; leaching the lignocellulosic feedstock by contacting the feedstock with at least one aqueous solution for a period greater than about 2 minutes to produce a leached feedstock and a leachate; removing the leachate from the leached feedstock; acidifying the leached feedstock to a pH between about 0.5 and about 3 to produce an acidified feedstock, and; reacting the acidified feedstock under conditions which disrupt fiber structure and hydrolyze a portion of hemicellulose and cellulose of the acidified feedstock, to produce a composition comprising xylose and a pretreated feedstock. The xylose may be purified from the pretreated feedstock or it may be converted to ethanol with the pretreated feedstock.

Abstract: A system for collecting ligno-cellulosic biomass over a large area to enable the commercial refining of biomass from 2,500 to in excess of 50,000 tons of biomass per day to produce ethanol or other products. The biomass is collected at a series of collection points and then transported through a network of conduit “loops” interconnecting each of the collection points and the central refining plant. The water used to transport the biomass, as a slurry, is recovered and sequentially recycled in the same pipeline system to push the biomass slurry around the system in a “loop.” The outgoing and return legs of each loop optionally are located adjacent each other.

Abstract: The present invention relates to novel xylose-fermenting yeast strains (for example, yeast of the genus Saccharomyces, e.g., S. cerevisiae) with an enhanced ability to ferment the xylose (and/or another pentose sugar) present in a lignocellulosic hydrolysate to a fermentation product(s) (for example, an alcohol (e.g., ethanol) or a sugar alcohol (e.g., xylitol)).

Abstract: The present invention provides a method of producing xylose from lignocellulosic feedstock. The method comprises disrupting lignocellulosic feedstock; leaching the lignocellulosic feedstock by contacting the feedstock with at least one aqueous solution for a period greater than about 2 minutes to produce a leached feedstock and a leachate; removing the leachate from the leached feedstock; acidifying the leached feedstock to a pH between about 0.5 and about 3 to produce an acidified feedstock, and; reacting the acidified feedstock under conditions which disrupt fiber structure and hydrolyze a portion of hemicellulose and cellulose of the acidified feedstock, to produce a composition comprising xylose and a pretreated feedstock. The xylose may be purified from the pretreated feedstock or it may be converted to ethanol with the pretreated feedstock.

Abstract: A modified Family 6 cellulase enzyme comprising a proline residue at position 413 is provided. Genetic constructs and genetically modified microbes comprising DNA sequences encoding the modified Family 6 cellulase are also provided. Family 6 cellulases of the invention display improved thermostability, thermophilicity, alkalophilicity, or a combination thereof, relative to the parent Family 6 cellulases. Such cellulases find use in a variety of applications in industry that require cellulase stability and activities at temperatures, pH values, or both, above that of the native enzyme.

Abstract: Provided are modified beta-glucosidase enzymes, derived from the Trichoderma reesei Cel3A beta-glucosidase, that exhibit improvements in one or more kinetic parameters (KG, KG2, kcat) comprising amino acid substitutions at one or more of positions 43, 101, 260 and 543. Also provided are genetic constructs comprising nucleotide sequences encoding for modified beta-glucosidase enzymes, methods for the production of modified beta-glucosidase enzymes from host strains and the use of the modified beta-glucosidase enzymes in the hydrolysis of cellulose.