Provided herein is a process for hydrolyzing a cellulosic feedstock to produce sugar. The process comprises introducing a pretreated cellulosic slurry to an inlet region of a plug flow hydrolysis reactor using a slurry introduction device that reduces the axial momentum of the slurry at the surface of the reactor contents. The cellulosic feedstock slurry is hydrolyzed in the plug flow hydrolysis reactor by contacting the cellulosic feedstock with at least cellulase enzymes to produce glucose. Also provided herein is a vertically-oriented, unmixed downflow hydrolysis reactor for hydrolyzing a pretreated cellulosic feedstock slurry which comprises such a slurry introduction device disposed in a top region thereof.

A method for treating a forage to increase digestibility by a ruminant comprises providing a forage for a ruminant comprising a level of ammonia that is from 0.0% to about 1.4% of dry matter (wt/wt). An alkali is applied at a dosage of from about 0.25% to about 0.75% of dry matter (wt/wt) to the forage. At least one exogenous carbohydrase enzyme is applied to the forage after application of the alkali and the forage is subsequently fed to the ruminant.

The present disclosure provides mutant cells for the secretion of proteins and for the degradation of lignocellulosic biomass. Methods for the use of these cells are also provided. Specifically, the utility of combined genetic deletions of β-glucosidases and the catabolite repressor gene creA/cre-1 for protein secretion in fungal and yeast cells is disclosed.

Provided is a method for recombinant expression of β-glucosidase gene. Also provided is a recombinant expression vector comprising: (a) a coding sequence of an aspartic protease or active fragment thereof, (b) a coding sequence of β-glucosidase or active fragment thereof, and optionally (c) a linker sequence between (a) and (b). Further provided are the recombinant host cell and the recombinant cellulose-degrading microorganism comprising the recombinant expression vector, the preparation method and uses thereof.

The present invention provides process for treating crop kernels, comprising the steps of a) soaking kernels in water to produce soaked kernels; b) grinding the soaked kernels; c) treating the soaked kernels in the presence of an effective amount of an enzyme composition comprising: i) a protease, and ii) a cellulolytic composition, wherein step c) is performed before, during or after step b).

Provided is a mutant β-glucosidase capable of more efficiently saccharifying biomass. A mutant β-glucosidase comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 1, wherein the amino acid sequence has asparagine at one or more positions selected from the group consisting of positions corresponding to positions 787, 790, and 797 of SEQ ID NO: 1, and has β-glucosidase activity.

The present invention provides ionic liquid-tolerant cellulases and method of producing and using such cellulases. The cellulases of the invention are useful in saccharification reactions using ionic liquid treated biomass.

The disclosure discloses the preparation of thermophilic β-glucosidase and the application thereof, which belongs to the technical field of genetic engineering and fermentation engineering. The present disclosure heterologously expresses the β-glucosidase TpBgl3A derived from thermophilic fungus Talaromyces piceae by constructing a recombinant bacteria. The enzyme production of recombinant bacteria can reach 2324 U/mL in a 3.6 L fermenter. The resulting β-glucosidase TpBgl3A can produce gentioligosaccharides with a high conversion rate using glucose as a substrate at a lower enzyme amount added, which significantly reduces production costs. In the reaction system using glucose and cellobiose as substrates, the conversion rate of gentioligosaccharide reaches 26.2%, which has the potential for industrial production of gentioligosaccharide.

Methods of preparing highly purified steviol glycosides, particularly rebaudiosides A, D and M are described. The methods include utilizing recombinant microorganisms for converting various staring compositions to target steviol glycosides. In addition, novel steviol glycosides reb D2, reb M2, and reb I are disclosed, as are methods of preparing the same. The highly purified rebaudiosides are useful as non-caloric sweetener in edible and chewable compositions such as any beverages, confectioneries, bakery products, cookies, and chewing gums.

The present disclosure relates to a method for preparing an aglycone or hydrolyzed glycoside converted from a glycoside and, specifically, to a method for preparing an aglycone or hydrolyzed glycoside from a glycoside by converting a glycoside into an aglycone form or hydrolyzed glycoside by using a microorganism producing β-glycosidase, and then recovering the aglycone or hydrolyzed glycoside accumulated in the cells of the microorganism.