title: Engineered biosynthesis and characterisation of disaccharide-modified 8-deoxyamphoteronolides.
abstract: Several polyene macrolides are potent antifungal agents that have severe side effects. Increased glycosylation of these compounds can improve water solubility and reduce toxicity. Three extending glycosyltransferases are known to add hexoses to the mycosaminyl sugar residues of polyenes. The Actinoplanes caeruleus PegA enzyme catalyses attachment of a D-mannosyl residue in a β-1,4 linkage to the mycosamine of the aromatic heptaene 67-121A to form 67-121C. NppY from Pseudonocardia autotrophica adds an N-acetyl-D-glucosamine to the mycosamine of 10-deoxynystatin. NypY from Pseudonocardia sp. P1 adds an extra hexose to a nystatin, but the identity of the sugar is unknown. Here, we express the nypY gene in Streptomyces nodosus amphL and show that NypY modifies 8-deoxyamphotericins more efficiently than C-8 hydroxylated forms. The modified heptaene was purified and shown to be mannosyl-8-deoxyamphotericin B. This had the same antifungal activity as amphotericin B but was slightly less haemolytic. Chemical modification of this new disaccharide polyene could give better antifungal antibiotics.
authors.affiliation: Department of Chemistry, University of Leicester, University Road, Leicester, LE1 7RH, UK.
authors.forename: Simon
authors.initials: S
authors.lastname: Walmsley
authors.affiliation: School of Biomolecular and Biomedical Science, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: Eimear
authors.initials: E
authors.lastname: De Poire
authors.affiliation: Department of Chemistry, University of Leicester, University Road, Leicester, LE1 7RH, UK.
authors.forename: Bernard
authors.initials: B
authors.lastname: Rawlings
authors.affiliation: School of Biomolecular and Biomedical Science, University College Dublin, Belfield, Dublin 4, Ireland. patrick.caffrey@ucd.ie.
authors.forename: Patrick
authors.identifier: http://orcid.org/0000-0002-0132-2410
authors.initials: P
authors.lastname: Caffrey
mesh_terms: D000117:Acetylglucosamine / Q000737:chemistry; D039903:Actinobacteria / Q000235:genetics / Q000378:metabolism; D000666:Amphotericin B / Q000009:adverse effects / Q000737:chemistry* / Q000494:pharmacology; D000935:Antifungal Agents / Q000009:adverse effects / Q000737:chemistry* / Q000494:pharmacology; D002176:Candida albicans / Q000187:drug effects*; D016695:Glycosyltransferases; D006595:Hexosamines / Q000737:chemistry; D007891:Leishmania / Q000187:drug effects*; D007896:Leishmaniasis / Q000188:drug therapy; D018942:Macrolides / Q000737:chemistry / Q000378:metabolism*; D009181:Mycoses / Q000188:drug therapy; D009761:Nystatin / Q000737:chemistry; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*
chemical_list: D000935:Antifungal Agents; D006595:Hexosamines; D018942:Macrolides; D009761:Nystatin; C014692:mycosamine; D000666:Amphotericin B; D016695:Glycosyltransferases; D000117:Acetylglucosamine
keywords: Amphotericin disaccharides; Glycosyltransferases; Streptomyces nodosus