The present invention relates to a method for producing trehalose, comprising the steps of mixing and reacting, in any order, (i) at least one alpha-phosphorylase capable of catalyzing the production of alpha-D-glucose 1-phosphate intermediate from a saccharide raw material, and from at least one phosphorus source; (ii) at least one trehalose phosphorylase capable of catalyzing the production of trehalose from an alpha-D-glucose 1-phosphate intermediate and a glucose substrate, wherein the trehalose phosphorylase is a trehalose phosphorylase variant with an amino acid sequence which differs from the amino acid sequence of a wild type trehalose phosphorylase in at least one amino acid position, (iii) at least one saccharide raw material which produces an alpha-D-glucose 1-phosphate intermediate and a co-product by catalytic action of the alpha-phosphorylase; and (iv) at least one phosphorus source selected from the group consisting of a phosphoric acids and an inorganic salt thereof.

The present invention provides engineered sucrose phosphorylase (SP) enzymes, polypeptides having SP activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. Methods for producing SP enzymes are also provided. The present invention further provides compositions comprising the SP enzymes and methods of using the engineered SP enzymes. The present invention finds particular use in the production of pharmaceutical compounds.

Provided herein are methods for the enzymatic production of beta-cyclodextrin from sucrose. In some cases, the methods involve contacting sucrose with one or more enzymes to convert sucrose to amylose, followed by contacting the amylose with one or more enzymes to convert the amylose to beta-cyclodextrin. In some cases, the methods produce higher yields of beta-cyclodextrin relative to alpha-cyclodextrin, gamma-cyclodextrin, or both.

The present invention relates to a method for producing a stable hydrogel-cell composition comprising the steps of: a) providing a reaction mixture comprising at least one polymerizable substrate and at least one enzyme capable of polymerizing said at least one substrate, and cells, and b) incubating the mixture of step a) to form a stable hydrogel-cell composition.

The present invention provides engineered sucrose phosphorylase (SP) enzymes, polypeptides having SP activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. Methods for producing SP enzymes are also provided. The present invention further provides compositions comprising the SP enzymes and methods of using the engineered SP enzymes. The present invention finds particular use in the production of pharmaceutical compounds.

Provided herein are methods for the enzymatic production of gamma-cyclodextrin from sucrose. In some cases, the methods involve contacting sucrose with one or more enzymes to convert sucrose to amylose, followed by contacting the amylose with one or more enzymes to convert the amylose to gamma-cyclodextrin. In some cases, the methods produce higher yields of gamma-cyclodextrin relative to alpha-cyclodextrin, beta-cyclodextrin, or both.

A method by which a cellooligosaccharide can be produced inexpensively at high percentage yield is provided. In the method for producing a cellooligosaccharide according to an embodiment, at least one primer selected from the group consisting of glucose, cellobiose, and derivatives in which anomeric positions of glucose and cellobiose are modified and sucrose are subjected to, in the presence of phosphoric acid, actions of sucrose phosphorylase and cellodextrin phosphorylase. In this case, in the reaction system, the concentration of phosphoric acid is set to 3 mol/m.sup.3 or more and 120 mol/m.sup.3 or less.