The present invention relates to trehalase variants of the Myceliophthora sepedonium GH37 trehalase. 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 disclosure concerns a recombinant yeast host cell exhibiting higher stability and, in some embodiments, higher fermentation performance. The recombinant yeast host cell stability has a limited ability to express an hydrolase during its propagation phase. In return, this limits the cleavage of a yeast cellular component during or after propagation which may be detrimental to the stability and/or fermentation performances. The recombinant yeast host cell expresses a heterologous hydrolase under the control of a heterologous promoter (for limiting the expression of the heterologous hydrolase during propagation and favoring the expression of the heterologous hydrolase during fermentation).

The present invention relates to genetically engineered yeasts having a heterologous trehalase gene and fermentation processes for using such yeasts. The yeasts can express trehalase in a quantity sufficient to convert significant amounts of trehalose to glucose, thereby improving the yield of the product in a fermentation, and/or reducing or eliminating the need to add exogenous trehalase to the fermentation. The yeasts can also include other heterologous genes for expressing enzymes useful for improving yield and/or for reducing or eliminating the need to add exogenous enzymes to the fermentation.

The present invention relates to genetically engineered yeasts having a heterologous trehalase gene and fermentation processes for using such yeasts. The yeasts can express trehalase in a quantity sufficient to convert significant amounts of trehalose to glucose, thereby improving the yield of the product in a fermentation, and/or reducing or eliminating the need to add exogenous trehalase to the fermentation. The yeasts can also include other heterologous genes for expressing enzymes useful for improving yield and/or for reducing or eliminating the need to add exogenous enzymes to the fermentation.

Described herein are recombinant fermenting organisms having a heterologous polynucleotide encoding an alpha-amylase and/or a heterologous polynucleotide encoding a trehalase. Also described are processes for producing a fermentation product, such as ethanol, from starch or cellulosic-containing material with the recombinant fermenting organisms.

The present invention relates to genetically engineered yeasts having a heterologous trehalase gene and fermentation processes for using such yeasts. The yeasts can express trehalase in a quantity sufficient to convert significant amounts of trehalose to glucose, thereby improving the yield of the product in a fermentation, and/or reducing or eliminating the need to add exogenous trehalase to the fermentation. The yeasts can also include other heterologous genes for expressing enzymes useful for improving yield and/or for reducing or eliminating the need to add exogenous enzymes to the fermentation.

The present disclosure relates to recombinant yeast host cells having (i) a first genetic modification for reducing the production of one or more native enzymes that function to produce glycerol or regulating glycerol synthesis and/or allowing the production of an heterologous glucoamylase and (ii) a second genetic modification for reducing the production of one or more native enzymes that function to produce trehalose or regulating trehalose synthesis and/or allowing the expression of an heterologous trehalase. The recombinant yeast host cells can be used to limit the production of (yeast-produced) trehalose (particularly extracellular trehalose) during fermentation and, in some embodiments, can increase the production of a fermentation product (such as, for example, ethanol).

The present disclosure relates to recombinant yeast host cells having (i) a first genetic modification for reducing the production of one or more native enzymes that function to produce glycerol or regulating glycerol synthesis and/or allowing the production of an heterologous glucoamylase and (ii) a second genetic modification for reducing the production of one or more native enzymes that function to produce trehalose or regulating trehalose synthesis and/or allowing the expression of an heterologous trehalase. The recombinant yeast host cells can be used to limit the production of (yeast-produced) trehalose (particularly extracellular trehalose) during fermentation and, in some embodiments, can increase the production of a fermentation product (such as, for example, ethanol).

The present disclosure relates to a method referred to herein as the split trehalase assay biosensor (also referred to herein as STIGA) is based on the use of engineered E. coli trehalase to detect analytes such as antibodies in a sample. The trehalase is engineered in a way such that the enzyme is split into two inactive fragments (N-terminal fragment H and C-terminal fragment A) with antigens fused to both fragments. When bivalent antibodies react specifically with the fused antigens, two inactive trehalase fragments are brought together in close proximity to restore the activity of trehalase. The restored trehalase will hydrolyze trehalose into two glucose molecules that can be measured using existing glucose detection methods such as glucometer, Benedict's reagent, or ACCU-CHEK AVIVA glucose test strips.

The present invention relates to trehalase variants of the Myceliophthora sepedonium GH37 trehalase. 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.