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).

Compositions and methods to treat biofilms are disclosed based on the discovery of the role of the disaccharide trehalose in microbial biofilm development. In various embodiments to treat body-borne biofilms systemically and locally, the method includes administering trehalase, the enzyme which degrades trehalose, in combination with other saccharidases for an exposition time sufficient to adequately degrade the biofilm gel matrix at the site of the biofilm. The method also includes administering a combination of other enzymes such as proteolytic, fibrinolytic, and lipolytic enzymes to break down proteins and lipids present in the biofilm, and administering antimicrobials for the specific type(s) of infectious pathogen(s) underlying the biofilm. Additionally, methods are disclosed to address degradation of biofilms on medical device surfaces and biofilms present in industrial settings.

The present invention relates to polypeptides having trehalase activity, particularly derived from Talaromyces. 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 for the production of ethanol.

Disclosed is a trehalose solution for use with urinary glucose measurement kits, in in vitro diagnostic (biochemical medical diagnosis) field, to measure urinary trehalase enzyme activity as a marker of kidney damage, and a measurement method where the solution is used.

Provided is an application of trehalase in fermentative production. The trehalase has amino acid sequences shown in SEQ ID NO.6, SEQ ID NO.7, and SEQ ID NO.8. Provided are methods for producing and applying trehalase, particularly being applied in the production and fermentation of alcohol and an amino acid.

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.

Compositions and methods to treat biofilms are disclosed based on the discovery of the role of the disaccharide trehalose in microbial biofilm development. In various embodiments to treat body-borne biofilms systemically and locally, the method includes administering trehalase, the enzyme which degrades trehalose, in combination with other saccharidases for an exposition time sufficient to adequately degrade the biofilm gel matrix at the site of the biofilm. The method also includes administering a combination of other enzymes such as proteolytic, fibrinolytic, and lipolytic enzymes to break down proteins and lipids present in the biofilm, and administering antimicrobials for the specific type(s) of infectious pathogen(s) underlying the biofilm. Additionally, methods are disclosed to address degradation of biofilms on medical device surfaces and biofilms present in industrial settings.

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 in close proximity to restore the activity of trehalase. The restored trehalase will hydrolyze trehalose into two glucose molecules which can be measured using existing glucose detection methods such as glucometer, Benedict's reagent, or ACCU-CHEK AVIVA? glucose test strips.

dsRNA generated from D. citri trehalase gene is effective in reducing fitness and/or survival of D. citri. Thus genetically altered plants expressing the dsRNA and plants to which dsRNA solutions are applied increase D. citri mortality and reduce D. citri infestation. With reduced D. citri population, the spread of microorganisms for which D. citri is a vector is reduced. Such microorganisms include, but are not limited to, C. Liberibacter species, including: CLas, CLam, and CLaf. Thus, applying of the D. citri trehalase dsRNA to a plant reduces disease and/or microorganism transmission by killing D. citri that feed on the treated plant.