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

摘要

The present invention relates to methods of use of glycosyltransferases and related novel compounds. The invention exploits the reversibility of glycosyltransferases to generate new sugars, unnatural biomolecules and numerous one-pot reactions for generation of new biomolecules having varied backbones such as enediynes, vancomycins, bleomycins, anthracyclines, macrolides, pluramycins, aureolic acids, indolocarbazoles, aminglycosides, glycopeptides, polyenes, coumarins, benzoisochromanequinones, calicheamicins, erythromycin, avermectins, ivermectins, angucyclines, cardiac glycosides, steroids or flavinoids. In preferred embodiments, the invention specifically relates to biosynthesis of anticancer (the enediyne calicheamicin, CLM), anthelmintic agents (the macrolides avermectin, ivermectin and erythromycin) and antibiotic (the glycopeptide vancomycin, VCM) natural product-based drugs developed by reversible, bidirectional glycosyltransferase-catalyzed reactions.

主权项

1. A method of generating a biomolecule A, in situ, having a sugar moiety X from a biomolecule B having the sugar moiety X, 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 sugar moiety X; and (b) isolating the biomolecule A having sugar moiety X from step (a), wherein the glycotransferase is selected from the group consisting of Micromonospora echinospora calicheamicin glycosyltransferase 1 (CalG1), Micromonospora echinospora calicheamicin glycosyltransferase 2 (CalG2), Micromonospora echinospora calicheamicin glycosyltransferas 3 (CalG3), Micromonospora echinospora calicheamicin glycosyltransferase 4 (CalG4), Amycolaotopsis orientalis glycosyltransferase D (GtfD), Amycolatopsis orientalis glycosyltransferase E (GtfE), Saccharopolyspora erythraea erythromycin glycosyltransferase BV (EryBv), and Streptmyces avermitilis ayermectin 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 henzoisochromancquinone, a calicheamicin, an erythromycin, an avermectin, an ivermectin, an angucycline, a cardiac glycoside a steroid or a flavinoid.