DANISCO US INC.

The instant disclosure is generally related to microbial fermentation, grain processing and ethanol production. Certain embodiments of the disclosure are therefore related to starch-containing materials and the microbial fermentation of such starch-containing materials for the production ethanol. More particularly, certain embodiments are related to novel protease mixtures for use in processing such starch-containing materials in starch to ethanol fermentation processes described herein.

Described are compositions and methods relating to a coated granule comprising a core coated with cross-linkable polymer, where the cross-linkable polymer is applied using a feed suspension that includes an acid-neutralized volatile base and a calcium salt insoluble at the pH of the neutralized feed suspension. Volatilization of the base decreases the pH of the feed suspension, thereby solubilizing the calcium salt, and inducing in situ ion-mediated cross-linking of the cross-linkable polymer.

Described is a process for releasing soluble starch hydrolysates or hydrolysed starch syrup from substantially intact corn kernels during high temperature steeping at a temperature at or above the gelatinization temperature of starch in the corn kernels, and the recovery of protein and oil-enhanced, carbohydrate-depleted residuals that are ideally suited for use as high quality animal feed or other food-grade products.

The present disclosure provides AprL-clade protease enzymes, including variant AprL-clade protease enzymes, nucleic acids encoding same, and compositions and methods related to the production and use thereof, including an AprL-clade variant subtilisin enzyme that has improved stability and/or soil removal compared to a parent AprL-clade subtilisin enzyme.

The present invention provides variant subtilisins and compositions comprising at least one variant subtilisin set forth herein, as well as methods for using these variants and compositions. In some embodiments, the present invention provides variant subtilisins suitable for laundry cleaning applications.

Certain embodiments of the disclosure are related to, inter alia, mutant and/or modified Bacillus cells (strains) comprising enhanced protein productivity phenotypes, mutant and/or modified cells comprising enhanced/increased ilvE messenger RNA (mRNA) levels, mutant and/or modified cells comprising enhanced/increased carbon yields (carbon yield efficiencies) of heterologous proteins produced and the like. As generally described herein, certain embodiments of the disclosure are related to mutant and/or modified Bacillus strains comprising variant ilvE genes, which mutant and/or modified Bacillus strains are particularly useful for the enhanced production of proteins of interests.

A method of identifying a heterologous polypeptide having enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA in cytosol of a yeast cell comprising: a) providing a mutated yeast cell comprising a deletion of at least one gene of the by-pass, selected from the genes encoding the enzymes pyruvate decarboxylase, acetaldehyde dehydrogenase, and acetylCoA synthetase; b) transforming said mutated yeast cell with an expression vector comprising a heterologous nucleotide sequence encoding a candidate polypeptide having potential enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA; c) testing said recombinant mutated yeast cell for its ability to grown on minimal medium containing glucose as sole carbon source, and d) identifying said candidate polypeptide as a heterologous polypeptide having enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA in the cytosol of said yeast cell when growth of said cell is observed.

Methods and compositions are provided for modifying the genome of Bacillus sp. cells without the use of a selectable marker and without the use of a guided Cas endonuclease. The disclosure includes methods for integrating donor DNA sequences into the genome of a Bacillus sp. cell without the use of a selectable marker and without the use of Cas endonucleases into said genome, as well as methods for deleting genes of interest and/or providing point mutations into the genome of Bacillus sp. cells.

The present disclosure is generally related to mutant and genetically modified filamentous fungal cells and methods thereof for use in the production of proteins of interest. More particularly, as described herein, the mutant and/or modified fungal cells (strains) of the disclosure are well-suited for use in industrial scale fermentation processes for the enhanced expression/production of proteins of interest in the absence and/or in the presence of an inducing substrate.

Described are compositions and methods relating to modified yeast that over-express the glycerol-deficient suppressor, GDS1. The yeast produces an increased amount of ethanol and a deceased amount of acetate compared to parental cells. Such yeast is particularly useful for large-scale ethanol production from starch substrates where acetate in an undesirable end product.