Glycorandomization and production of novel vancomycin analogs
The present invention provides combinatorial methods for rapidly generating a diverse library of glycorandomized structures, comprising incubating one or more aglycons and a pool of NDP-sugars in the presence of a glycosyltransferase. The glycosyltransferase may be one that is associated with or involved in production of natural secondary metabolites, or one which is putatively associated with or involved in production of natural secondary metabolites. The glycosyltransferase may show significant flexibility with respect to its NDP-sugar donors and/or its aglycons. NDP-sugar donors may be commercially available, or may be produced by utilizing mutant or wild type nucleotidyltransferases significant flexibility with respect to their substrates.
1. A method of producing a chemoselectively ligated vancomycin analog comprising incubating at least one glycosylated vancomycin comprising a glycosyl moiety selected from the group consisting of:
with a chemoselectively ligatable agent comprising an alkyne moiety, wherein the glycosyl moiety and chemoselectively ligatable agent react to provide a chemoselectively ligated vancomycin analog having a structure selected from the group consisting of:
wherein at least one chemoselectively ligated vancomycin analog is produced.
2. A method according to claim 1 , further, wherein the glycosylated vancomycin is initially produced by incubating vancomycin and at least one thymidine or uridine nucleotide diphosphosugar comprising a sugar structure selected from the group consisting of:
in the presence of at least one first glycosyltransferase wherein the at least glycosylated vancomycin is produced.