The present invention relates to cellobiohydrolase variants and carbohydrate binding module variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

The present invention relates to isolated polypeptides having cellobiohydrolase activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

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).

The present invention relates to cellobiohydrolase variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

The present invention relates to cellobiohydrolase variants and carbohydrate binding module variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

A heterologous gene cassette useful for creating Streptomyces species with enhanced capability of growing on a cellulosic polysaccharide substrate, wherein the cassette comprises at least two members of the following categories: a) a GH6 gene, b) an AA10 gene, c) a GH48 gene, d) a GH5 gene and e) either (i) a GH9 gene, (ii) a GH9 gene and a GH12 gene, or (iii) a GH12 gene is disclosed.

A thermostable cellobiohydrolase, having a cellobiohydrolase catalytic domain including: (A) a polypeptide including the amino acid sequence represented by SEQ ID NO: 1, (B) a polypeptide including an amino acid sequence in which at least one amino acid has been deleted, substituted, or added in the amino acid sequence represented by SEQ ID NO: 1, and having hydrolysis activity against a substrate of phosphoric acid swollen Avicel at least under conditions of 65° C. and pH 6, or (C) a polypeptide including an amino acid sequence having 80% or greater sequence identity with the amino acid sequence represented by SEQ ID NO: 1, and having hydrolysis activity against a substrate of phosphoric acid swollen Avicel at least under conditions of 65° C. and pH 6.

A thermostable cellobiohydrolase, having a cellulose-binding motif domain including (A1) a polypeptide including an amino acid sequence represented by SEQ ID NO: 1, (B1) a polypeptide including an amino acid sequence in which at least one amino acid has been deleted, substituted, or added in the amino acid sequence represented by SEQ ID NO: 1, and having a cellulose-binding function, or (C1) a polypeptide including an amino acid sequence having 70% or greater sequence identity with the amino acid sequence represented by SEQ ID NO: 1, and having a cellulose-binding function, and also having a cellobiohydrolase catalytic domain, wherein the thermostable cellobiohydrolase exhibits hydrolysis activity against a substrate of phosphoric acid swollen Avicel at least under conditions of 95° C. and pH 5.5.

The present invention is directed to a yeast strain, or strains, secreting a full suite, or any subset of that full suite, of enzymes to hydrolyze corn starch, corn fiber, lignocellulose, (including enzymes that hydrolyze linkages in cellulose, hemicellulose, and between lignin and carbohydrates) and to utilize pentose sugars (xylose and arabinose). The invention is also directed to the set of proteins that are well expressed in yeast for each category of enzymatic activity. The resulting strain, or strains can be used to hydrolyze starch and cellulose simultaneously. The resulting strain, or strains can be also metabolically engineered to produce less glycerol and uptake acetate. The resulting strain, or strains can also be used to produce ethanol from granular starch without liquefaction. The resulting strain, or strains, can be further used to reduce the amount of external enzyme needed to hydrolyze a biomass feedstock during an Simultaneous Saccharification and Fermentation (SSF) process, or to increase the yield of ethanol during SSF at current saccharolytic enzyme loadings. In addition, multiple enzymes of the present invention can be co-expressed in cells of the invention to provide synergistic digestive action on biomass feedstock. In some aspects, host cells expressing different heterologous saccharolytic enzymes can also be co-cultured together and used to produce ethanol from biomass feedstock.

A strategy for eliminating or greatly reducing the need for physical/chemical treatments or the use of whole microbes for lignocellulosic biomass and organopollutant degradation is disclosed. The soybean is a practical, cost-efficient and sustainable bioreactor for the production of lignin-degrading and cellulose-degrading enzymes. The use of soybean as a transgenic overexpression platform provides advantages that no other industrial scale enzyme expression system can match. Availability of a battery of related plant biomass degrading enzymes in separate transgenic soybean lines provides unprecedented flexibility in industrial and bioremediation processes. Depending upon the particular application, selected soybean-derived powdered enzyme formulations can be used, and their sequential addition can be orchestrated. Manufacturing enzymes using transgenic soybeans wherein these enzymes are capable of lignocellulose and organopollutant degradation into useful or nontoxic products will dramatically change biomass engineering schemes and environmental remediation practices. This technology has a sum of advantages that other protein expression system cannot duplicate, including the manufacturing of individual enzymes in a cost-effective manner that allows flexibility in cocktail composition, ease of application, and long term storage in the absence of a cold chain.