The membrane bioreactor for simultaneous enzymatic cellulose hydrolysis and product separation is a vessel having a lower hydrolysis reaction chamber and an upper distilled water chamber separated by a semipermeable membrane attached to the top of the lower reaction chamber. The membrane is supported on a stainless steel mesh and sealed to the mesh by epoxy glue to prevent leakage. A peristaltic pump is connected to the reaction chamber and maintains a flow of distilled water through the membrane and the upper chamber, the effluent being collected in a beaker or other product collection vessel. The reaction chamber is agitated at a moderate rate by a magnetic stirrer, and the upper chamber is agitated more rigorously by a mechanical stirrer. A thermocouple and temperature controller and a buffer solution, respectively, maintain temperature and pH in the reaction chamber optimal for enzymatic hydrolysis of cellulose.

The present invention provides an endoglucanase having excellent heat resistance. Specifically, the present invention provides an endoglucanase satisfying characteristics (A) and (B) below: (A) having an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 1; and (B) having at least one amino acid substitution selected from the group consisting of K214E, D254E, and S309P.

The present invention provides an endoglucanase having excellent heat resistance. Specifically, the present invention provides an endoglucanase satisfying characteristics (A) and (B) below: (A) having an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 1; and (B) having at least one amino acid substitution selected from the group consisting of K214E, D254E, and S309P.

The present disclosure provides methods for generating sugars from a cellulosic biomass. The methods combine treatment of the biomass using a high-shear milling device and saccharification of the biomass to partially hydrolyze the biomass. The biomass can be saccharified either after or simultaneously with the high-shear milling treatement. The partially hydrolyzed biomass is then separated into a solids stream with saccharification enzymes, and a liquid stream with sugars. The solids stream and associated enzymes are further incubated under saccharification conditions to produce additional sugars, or are recycled and added to fresh biomass, which is saccharified under high-shear milling conditions. The methods result in improved conversion of cellulosic biomass to glucose.

The present disclosure provides methods for generating sugars from a cellulosic biomass. The methods combine treatment of the biomass using a high-shear milling device and saccharification of the biomass to partially hydrolyze the biomass. The biomass can be saccharified either after or simultaneously with the high-shear milling treatement. The partially hydrolyzed biomass is then separated into a solids stream with saccharification enzymes, and a liquid stream with sugars. The solids stream and associated enzymes are further incubated under saccharification conditions to produce additional sugars, or are recycled and added to fresh biomass, which is saccharified under high-shear milling conditions. The methods result in improved conversion of cellulosic biomass to glucose.

A Trichoderma reesei mutant strain has a function of a polypeptide consisting of the amino acid sequence represented by SEQ ID NO: 2 is reduced. A method of producing a protein includes a step of cultivating the Tricho-derma reesei mutant strain, and a method of producing a cellulase includes a step of cultivating the Trichoderma reesei mutant strain.

A technique for processing ancient, heritage and modern wheat, grains, seeds, beans, legumes, tuber and root vegetables create baking flours suitable for human consumption. The initial ingredient is incubated to initiate germination and activate internal enzymes and nutrient production for useful enzymes, proteins and nutrients. Germination is terminated and the product wet-milled to fracture or shear the outer hull, exposing the inner grain. The product is mixed with water at varying temperatures during which amylase is added. The mixture is incubated to facilitate saccharification of starches into sugars by the amylase enzymes. The mixture is pasteurized to denature the amylases and the mash pressed and/or strained to separate the liquid and solids. The solid phase is dried and milled into higher fiber, high protein, low carbohydrate flour. The liquid is carbohydrate-rich with substantial fiber, protein and other nutrients dissolved in the solution.

A technique for processing ancient, heritage and modern wheat, grains, seeds, beans, legumes, tuber and root vegetables create baking flours suitable for human consumption. The initial ingredient is incubated to initiate germination and activate internal enzymes and nutrient production for useful enzymes, proteins and nutrients. Germination is terminated and the product wet-milled to fracture or shear the outer hull, exposing the inner grain. The product is mixed with water at varying temperatures during which amylase is added. The mixture is incubated to facilitate saccharification of starches into sugars by the amylase enzymes. The mixture is pasteurized to denature the amylases and the mash pressed and/or strained to separate the liquid and solids. The solid phase is dried and milled into higher fiber, high protein, low carbohydrate flour. The liquid is carbohydrate-rich with substantial fiber, protein and other nutrients dissolved in the solution.

The present invention relates to an improved method and device for treating biomass in which thermally treated biomass is discharged from a pressurized reactor and introduced into a blow tank, wherein the absolute pressure in the blow tank is maintained below atmospheric pressure. The slurry of biomass separated in the blow tank is then enzymatically treated.

The present invention relates to an improved method and device for treating biomass in which thermally treated biomass is discharged from a pressurized reactor and introduced into a blow tank, wherein the absolute pressure in the blow tank is maintained below atmospheric pressure. The slurry of biomass separated in the blow tank is then enzymatically treated.