title: Redesign of polyene macrolide glycosylation: engineered biosynthesis of 19-(O)-perosaminyl-amphoteronolide B.
abstract: Most polyene macrolide antibiotics are glycosylated with mycosamine (3,6-dideoxy-3-aminomannose). In the amphotericin B producer, Streptomyces nodosus, mycosamine biosynthesis begins with AmphDIII-catalyzed conversion of GDP-mannose to GDP-4-keto-6-deoxymannose. This is converted to GDP-3-keto-6-deoxymannose, which is transaminated to GDP-mycosamine by the AmphDII protein. The glycosyltransferase AmphDI transfers mycosamine to amphotericin aglycones (amphoteronolides). The aromatic heptaene perimycin is unusual among polyenes in that the sugar is perosamine (4,6-dideoxy-4-aminomannose), which is synthesized by direct transamination of GDP-4-keto-6-deoxymannose. Here, we use the Streptomyces aminophilus perDII perosamine synthase and perDI perosaminyltransferase genes to engineer biosynthesis of perosaminyl-amphoteronolide B in S. nodosus. Efficient production required a hybrid glycosyltransferase containing an N-terminal region of AmphDI and a C-terminal region of PerDI. This work will assist efforts to generate glycorandomized amphoteronolides for drug discovery.
authors.affiliation: School of Biomolecular and Biomedical Science and Centre for Synthesis and Chemical Biology, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: Eve
authors.initials: E
authors.lastname: Hutchinson
authors.forename: Barry
authors.initials: B
authors.lastname: Murphy
authors.forename: Terence
authors.initials: T
authors.lastname: Dunne
authors.forename: Ciaran
authors.initials: C
authors.lastname: Breen
authors.forename: Bernard
authors.initials: B
authors.lastname: Rawlings
authors.forename: Patrick
authors.initials: P
authors.lastname: Caffrey
mesh_terms: D000666:Amphotericin B / Q000031:analogs & derivatives* / Q000096:biosynthesis; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology; D001483:Base Sequence; D002238:Carbohydrate Epimerases / Q000235:genetics / Q000378:metabolism; D015195:Drug Design; D005798:Genes, Bacterial; D005818:Genetic Engineering; D006031:Glycosylation; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism; D018942:Macrolides / Q000737:chemistry* / Q000494:pharmacology; D008969:Molecular Sequence Data; D005810:Multigene Family; D011090:Polyenes / Q000737:chemistry*; D013302:Streptomyces / Q000201:enzymology; D000637:Transaminases / Q000235:genetics / Q000378:metabolism
chemical_list: C549790:19-(O)-perosaminyl-amphoteronolide B; D000900:Anti-Bacterial Agents; D018942:Macrolides; D011090:Polyenes; D000666:Amphotericin B; D016695:Glycosyltransferases; D000637:Transaminases; D002238:Carbohydrate Epimerases; C061272:perosamine synthetase