Provided are methods for protein engineering, such as engineering proteases or kinases. The methods may utilize yeast display and/or ER sequestration of proteins or substrates. In some aspects, TEV proteases with altered substrate specificity, potency, and/or efficiency are provided.

Recombinant nucleic acid constructs for expression of heterologous cytokines or chemokines in C. albicans, and genetically modified C. albicans strains comprising the recombinant nucleic acid constructs, are described. The recombinant nucleic acid molecules include a C. albicans gene promoter sequence, a nucleic acid sequence encoding a C. albicans secreted protein signal sequence and a heterologous open reading frame (ORF) of a cytokine or chemokine gene. Also described are a method of expressing a heterologous cytokine or chemokine protein in a subject, and a method of treating or inhibiting the development of candidiasis in a subject. These methods include administering a genetically modified C. albicans containing a recombinant nucleic acid construct for expression of a heterologous cytokine or chemokine. Exemplary cytokines or chemokines include IL8, IL17A, CXCL1, CXCL2 and CXCL5.

Recombinant nucleic acid constructs for expression of heterologous cytokines or chemokines in C. albicans, and genetically modified C. albicans strains comprising the recombinant nucleic acid constructs, are described. The recombinant nucleic acid molecules include a C. albicans gene promoter sequence, a nucleic acid sequence encoding a C. albicans secreted protein signal sequence and a heterologous open reading frame (ORF) of a cytokine or chemokine gene. Also described are a method of expressing a heterologous cytokine or chemokine protein in a subject, and a method of treating or inhibiting the development of candidiasis in a subject. These methods include administering a genetically modified C. albicans containing a recombinant nucleic acid construct for expression of a heterologous cytokine or chemokine. Exemplary cytokines or chemokines include IL8, IL17A, CXCL1, CXCL2 and CXCL5.

The invention relates to the field of bakery ingredients. More specifically, the invention relates to a variant of a parent polypeptide, wherein the difference between the variant polypeptide and the parent polypeptide is that a KEX2 protease cleavage site which is present in the parent polypeptide, is absent in the variant polypeptide. The invention further relates to a process for preparing a dough wherein a variant polypeptide as disclosed herein is used and baked product prepared from the dough.

The invention relates to the field of bakery ingredients. More specifically, the invention relates to a variant of a parent polypeptide, wherein the difference between the variant polypeptide and the parent polypeptide is that a KEX2 protease cleavage site which is present in the parent polypeptide, is absent in the variant polypeptide. The invention further relates to a process for preparing a dough wherein a variant polypeptide as disclosed herein is used and baked product prepared from the dough.

The present disclosure relates to proteases for improving alcoholic fermentation. The proteases are expressed from a recombinant host cell. The present disclosure also provides a population of recombinant host cells expressing an heterologous protease that can be used in combination with recombinant host cells expressing an heterologous glucoamylase and/or an heterologous glycerol reduction system.

The present disclosure relates to proteases for improving alcoholic fermentation. The proteases are expressed from a recombinant host cell. The present disclosure also provides a population of recombinant host cells expressing an heterologous protease that can be used in combination with recombinant host cells expressing an heterologous glucoamylase and/or an heterologous glycerol reduction system.

The present disclosure concerns a process for fermenting a biomass with a reduced dose of a purified exogenous enzyme (which can be, for example a purified exogenous glucoamylase). The process comprises contacting a biomass (which may comprise starch) with a recombinant yeast host cell. The recombinant yeast host cell has a genetic modification for expressing a heterologous polypeptide having starch or dextrin hydrolase activity (which may be, for example, from a glucoamylase). The nucleic acid molecule encoding the heterologous polypeptide having starch or dextrin hydrolase activity comprises allowing the secretion of the heterologous polypeptide.

The present disclosure concerns a process for fermenting a biomass with a reduced dose of a purified exogenous enzyme (which can be, for example a purified exogenous glucoamylase). The process comprises contacting a biomass (which may comprise starch) with a recombinant yeast host cell. The recombinant yeast host cell has a genetic modification for expressing a heterologous polypeptide having starch or dextrin hydrolase activity (which may be, for example, from a glucoamylase). The nucleic acid molecule encoding the heterologous polypeptide having starch or dextrin hydrolase activity comprises allowing the secretion of the heterologous polypeptide.

Provided herein, inter alia, are fusion DNA constructs comprising improved protease recognition sequences for expressing and purifying one or more polypeptides of interest as well as methods for producing one or more polypeptides of interest in a recombinant host cell.