The present invention provides for a mechanism to reduce glycerol production and increase nitrogen utilization and ethanol production of recombinant microorganisms. One aspect of this invention relates to strains of S. cerevisiae with reduced glycerol productivity that get a kinetic benefit from higher nitrogen concentration without sacrificing ethanol yield. A second aspect of the invention relates to metabolic modifications resulting in altered transport and/or intracellular metabolism of nitrogen sources present in corn mash.

The present invention relates to a fusion protein, comprising the structure N-PCS.sup.Y-degSig.sub.N-M-PCS.sup.X-degSig.sub.CC; wherein N represents the N-terminus; PCS.sup.Y and PCS.sup.X each represent a protease cleavage site (PCS), which differ from each other in at least one amino acid residue; degSig.sub.N represents a degradation signal which promotes degradation of the fusion protein in a host pot cell if PCS.sup.Y is cleaved by a protease such that the first amino acid of degSig.sub.N becomes the new N-terminus of the remaining fusion; M represents a cytoplasmic selection marker; and degSig.sub.C represents a second degradation signal which promotes degradation of the fusion protein in a host cell if PCS.sup.X is not cleaved by a protease; and C represents the C-terminus. Further provided is a nucleic acid construct, comprising a nucleic acid sequence coding for said fusion protein, a nucleic acid expression construct library, comprising a plurality of such nucleic acid expression constructs in diversified form, and methods using the fusion protein and nucleic acid constructs coding therefor. Finally, the present invention provides variants of bdSUMO and bdSENP1 which have been identified by the methods of the present disclosure, and which exhibit improved properties over existing orthogonal protease/protease cleavage site-pairs which are currently used with wild-type bdSUMO and wildtype bdSENP1.

The present invention relates to a fusion protein, comprising the structure N-PCS.sup.Y-degSig.sub.N-M-PCS.sup.X-degSig.sub.CC; wherein N represents the N-terminus; PCS.sup.Y and PCS.sup.X each represent a protease cleavage site (PCS), which differ from each other in at least one amino acid residue; degSig.sub.N represents a degradation signal which promotes degradation of the fusion protein in a host pot cell if PCS.sup.Y is cleaved by a protease such that the first amino acid of degSig.sub.N becomes the new N-terminus of the remaining fusion; M represents a cytoplasmic selection marker; and degSig.sub.C represents a second degradation signal which promotes degradation of the fusion protein in a host cell if PCS.sup.X is not cleaved by a protease; and C represents the C-terminus. Further provided is a nucleic acid construct, comprising a nucleic acid sequence coding for said fusion protein, a nucleic acid expression construct library, comprising a plurality of such nucleic acid expression constructs in diversified form, and methods using the fusion protein and nucleic acid constructs coding therefor. Finally, the present invention provides variants of bdSUMO and bdSENP1 which have been identified by the methods of the present disclosure, and which exhibit improved properties over existing orthogonal protease/protease cleavage site-pairs which are currently used with wild-type bdSUMO and wildtype bdSENP1.

The present invention is directed to antibodies and fragments thereof and humanized versions thereof having binding specificity for IL-6. Another embodiment of this invention relates to the antibodies described herein, and binding fragments thereof, comprising the sequences of the V.sub.H, V.sub.L and CDR polypeptides described herein, and the polynucleotides encoding them. The invention also contemplates conjugates of anti-IL-6 antibodies and binding fragments thereof conjugated to one or more functional or detectable moieties. The invention also contemplates methods of making said anti-IL-6 antibodies and binding fragments thereof. Embodiments of the invention also pertain to the use of anti-IL-6 antibodies, and binding fragments thereof, for the diagnosis, assessment and treatment of diseases and disorders associated with IL-6. These antibodies may bind at least one of soluble IL-6, cell surface expressed IL-6, IL-6/IL-6R and/or prevent the association of IL-6 and IL-6R, the association of IL-6/IL-6R and gp130 and or the formation of IL-6/IL-6R/gp130 multimers and thereby inhibit a biological effect associated with any of the foregoing.

The present invention is directed to antibodies and fragments thereof and humanized versions thereof having binding specificity for IL-6. Another embodiment of this invention relates to the antibodies described herein, and binding fragments thereof, comprising the sequences of the V.sub.H, V.sub.L and CDR polypeptides described herein, and the polynucleotides encoding them. The invention also contemplates conjugates of anti-IL-6 antibodies and binding fragments thereof conjugated to one or more functional or detectable moieties. The invention also contemplates methods of making said anti-IL-6 antibodies and binding fragments thereof. Embodiments of the invention also pertain to the use of anti-IL-6 antibodies, and binding fragments thereof, for the diagnosis, assessment and treatment of diseases and disorders associated with IL-6. These antibodies may bind at least one of soluble IL-6, cell surface expressed IL-6, IL-6/IL-6R and/or prevent the association of IL-6 and IL-6R, the association of IL-6/IL-6R and gp130 and or the formation of IL-6/IL-6R/gp130 multimers and thereby inhibit a biological effect associated with any of the foregoing.

A vaccine is disclosed that promotes CD4+ T cell-independent host defense mechanisms to defend against infection by fungi such as Pneumocystis spp. The vaccine may be used to prevent or to treat fungal infections. The novel vaccine can provide protective immunity, even for immunocompromised individuals such as HIV patients having reduced levels of CD4+ T cells.

A vaccine is disclosed that promotes CD4+ T cell-independent host defense mechanisms to defend against infection by fungi such as Pneumocystis spp. The vaccine may be used to prevent or to treat fungal infections. The novel vaccine can provide protective immunity, even for immunocompromised individuals such as HIV patients having reduced levels of CD4+ T cells.

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.

The present invention relates to a method for preparing pregabalin by a biological enzyme method. In particular, the method comprises producing pregabalin B and an R-configuration compound C by using a compound A as a raw material under the action of a biological enzyme; performing configuration inversion of the separated and recovered R-configuration compound C under the action of an isomerase to produce an S-configuration compound D; and producing pregabalin B from the compound D under the action of a biological enzyme

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