The present disclosure relates a polypeptide having UGT activity, which polypeptide comprises an amino acid sequence which, when aligned with a polypeptide having UGT activity comprising the sequence set out in SEQ ID NO: 2, comprises at least one substitution of an amino acid corresponding to any of amino acids at positions 35, 189, 280, 284, 285, 334 or 373, said positions being defined with reference to SEQ ID NO: 2 and wherein the polypeptide has one or more modified properties as compared with a reference polypeptide having UGT activity. A polypeptide according to the disclosure may be used in a recombinant cell for the production of steviol or a steviol glycoside.

The invention provides methods for glycosylation and preparation of compounds. The compounds include galactosylated, sialylated, fucosylated, and N-acetylglucosaminylated compounds from simple animal-derived, plant-derived, or microbe-derived oligosaccharides and sugars. In certain embodiments, the invention provides trinucleotide-free enzymatic production of oligosaccharides starting from simple sugars that include plant-based sugars. The invention also provides the enzymatic production of fucosylated oligosaccharides and fucosylated antibody-glycan conjugates from common sugars. The production may be a cell-free, one-pot synthesis using enzymes, and in some embodiments, immobilized enzymes. The synthesis is a highly customizable and highly efficient cell-free manufacturing process. In some embodiments, lactose derivatives and human milk oligosaccharides (HMOs) are produced.

The present invention provides engineered glycosyltransferase (GT) enzymes, polypeptides having GT activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. The present invention provides engineered sucrose synthase (SuS) enzymes, polypeptides having SuS activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. The present invention also provides compositions comprising the GT enzymes and methods of using the engineered GT enzymes to make products with -glucose linkages. The present invention further provides compositions and methods for the production of rebaudiosides (e.g., rebaudioside M, rebaudioside A, rebaudioside I, and rebaudioside D). The present invention also provides compositions comprising the SuS enzymes and methods of using them. Methods for producing GT and SuS enzymes are also provided.

The present invention provides engineered glycosyltransferase (GT) enzymes, polypeptides having GT activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. The present invention provides engineered sucrose synthase (SuS) enzymes, polypeptides having SuS activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. The present invention also provides compositions comprising the GT enzymes and methods of using the engineered GT enzymes to make products with -glucose linkages. The present invention further provides compositions and methods for the production of rebaudiosides (e.g., rebaudioside M, rebaudioside A, rebaudioside I, and rebaudioside D). The present invention also provides compositions comprising the SuS enzymes and methods of using them. Methods for producing GT and SuS enzymes are also provided.

Heparosan synthase variants having improved expression levels, enhanced thermal stability, and/or reduced reverse glycosylation activity are provided. Methods for making oligosaccharides and polysaccharides, including heparin analogs and heparan sulfate analogs, are also described.

Methods of preparing highly purified steviol glycosides, particularly rebaudiosides A, D and Mare described. The methods include utilizing recombinant microorganisms for converting various staring compositions to target steviol glycosides. In addition, novel steviol glycosides reb D2, reb M2, and reb I are disclosed, as are methods of preparing the same. The highly purified rebaudiosides are useful as non-caloric sweetener in edible and chewable compositions such as any beverages, confectioneries, bakery products, cookies, and chewing gums.

The present invention provides host cells and methods for making mogrol glycosides, including Mogroside V (Mog. V), Mogroside VI (Mog. VI), Iso-Mogroside V (Isomog. V), and glycosylation products that are minor products in Siraitia grosvenorii. The invention provides engineered enzymes and engineered host cells for producing mogrol glycosylation products, such as Mog, V. Mog. VI, and Isomog. V, at high purity and/or yield. The present technology further provides methods of making products containing mogrol glycosides, such as Mog. V, Mog. VI, and Isomog. V, including food products, beverages, oral care products, sweeteners, and flavoring products.

An enzymatically produced soluble -glucan fiber composition is provided suitable for use as a digestion resistant fiber in food and feed applications. The soluble -glucan fiber composition can be blended with one or more additional food ingredients to produce fiber-containing compositions. Methods for the production and use of compositions comprising the soluble -glucan fiber are also provided.

The present invention relates to a host cell comprising an rhlA gene or an ortholog thereof that is capable of producing hydroxyalkanoyloxy alkanoic acid (HAA) and achieving an HAA concentration of more than 1 g L.sup.1 when cultured. The invention further relates to methods of producing such a host cell and to the use of said host cell for producing HAA. The present invention also relates to methods of producing HAA using said host cell, HAA compositions produced by these methods, as well as methods of producing fatty acid compositions, fatty alcohol compositions, or hydrocarbon compositions comprising producing HAA using said host cell, and fatty acid compositions, fatty alcohol compositions, or hydrocarbon compositions produced by said methods.

Provided is a method for preparing rebaudioside N using an enzymatic method, comprising using rebaudioside A or rebaudioside J as a substrate, and making the substrate, in the presence of a glycosyl donor, react under the catalysis of a UDP-glycosyl-transferase and/or a UDP-glycosyltransferase-containing recombinant cell to generate rebaudioside N.