Reversible natural product glycosyltransferase-catalyzed reactions, compounds and related methods

A method of generating a biomolecule A having a sugar moiety X and a biomolecule B having a sugar moiety Y from a hiomolecule B having the sugar moiety X and a biomolecule A having the sugar moiety Y comprising the steps of:

• (a) incubating the biomolecule A having the sugar moiety Y, biomolecule B having the sugar moiety X, and a nucleotide diphosphate in the presence of a glycosyltransferase wherein
• (i) the sugar inoiety X of the biomolecule B is excised from the biomolecule B, thereby generating a sugar moiety X-nucleotide diphosphate and a biomolecule aglycon B;
• (ii) the sugar moiety Y of the biomolecule A is excised from the biomolecule A, thereby generating sugar moiety Y-nucieotide diphosphate and a biomolecule aglycon A;
• and
• (iii) the sugar moiety X-nueleotide diphosphate and the biomolecule A are ligated and the sugar moiety Y-nueleotide diphosphate and the biomolecule B are ligated, thereby generating the biomolecule A having the sugar moiety X and biomolecule B having the sugar moiety Y;
• and
• (b) isolating the biomoleeule A having the sugar moiety X and the biomolecule B having from the sugar moiety Y from step (a)(iii),
• wherein the glygotransferase is selected from the group consisting of Micromonospora echlnospora calicheamicin glycosyltransferase 1 (CalG1), Micromonospora echinospora calicheamicin glycosyltransferase 2 (CalG2), Micromonospora echinspora calicheamicin glycosyltransferase 3 (CalG3), Micromonospora echinospora calicheamicin glycosyltransferase 4 (CalG4), Amycolatopsis orientalis glycosyltransferase D (GtfD), Amycolatopsis orientalis glycosyltransferase E (GtfE), Saccharopolvspora erythraea erythromycin glycosyltransferase BV (EryBV), and Streptomyces avermitilis avermectin glycosyltransferase (AveBI), and
• wherein the biomolecule is an enediyne, a vancomycin, a bleomycin, an anthracycline, a macrolide, a pluramycin, an aureolic acid, an indolocarbazole, an aminglycoside, a glycopeptide, a polyene, a coumarin a benzoisochromanequinone, a calicheamicin, an erythromycin, an avermectin, an ivermectin, an angueycline, a cardiac glycoside, a steroid or a flavinoid.

A method of generating a library of isolated glycosylated biomolecules comprising a biomolecule A having sugar moiety X, wherein the method comprises transferring the sugar moiety X from a biomolecule B having sugar moiety X (a natural glycoside biomolecule or a natural biomolecule having sugar moiety X) to the biomolecule A, the method comprising the steps of:

• (a) incubating the biomolecule A, the biomolecule B and a nucleotide diphosphate in the presence of a glycosyltransferase wherein
• (i) the sugar moiety X of the biomolecule B is excised from the biomolecule B, thereby generating an independent sugar moiety X and a biomolecule aglycon B, whereby a nucleotide diphosphate (NDP)-sugar intermediate is formed;
• and
• (ii) the independent sugar moiety X and the biomolecule A are ligated, thereby generating the biomolecule A having the sugar moiety X;
• and
• (b) isolating the biomolecule A having sugar moiety X from step (a),
• wherein glycosyltransferase selected from the group consisting of Micromonospora echinospora calicheamicin glycosyltransferase 1 (CalG1), Micromonospora echinospora calicheamicin glycosyltransferase 2 (CalG2), Micromonospora echinospora calicheamicin glycosyltransferase 3 (CalG3), Micromonospora echinospora calicheamicin glycosyltransferase 4 (CalG4), Amvcolatopsis orientalis glycosyltransferase D (GtfD), Amvcolatopsis orientalis glycosyltransferase E (GtfE), Saccharopolwpora erythraea erythromycin glycosyltransferase BV (EryBV), and Streptomvces avermitilis avermectin glycosyltransferase (AveBI) and
• wherein the biomolecule is an enediyne, a vancomycin, a bleomycin, an anthracycline, a macrolide, a pluramycin, an aureolic acid, an indolocarbazole, an aminglycoside, a glycopeptide, a polyene, a coumarin, a benzoisochromanequinone, a calicheamicin, an erythromycin, an avermectin, an ivermectin, an anguecycline, a cardiac glycoside, a steroid or a flavonoid.