title: Myxovirescin A biosynthesis is directed by hybrid polyketide synthases/nonribosomal peptide synthetase, 3-hydroxy-3-methylglutaryl-CoA synthases, and trans-acting acyltransferases.
abstract: Myxococcus xanthus DK1622 is shown to be a producer of myxovirescin (antibiotic TA) antibiotics. The myxovirescin biosynthetic gene cluster spans at least 21 open reading frames (ORFs) and covers a chromosomal region of approximately 83 kb. In silico analysis of myxovirescin ORFs in conjunction with genetic studies suggests the involvement of four type I polyketide synthases (PKSs; TaI, TaL, TaO, and TaP), one major hybrid PKS/NRPS (Ta-1), and a number of monofunctional enzymes similar to the ones involved in type II fatty-acid biosynthesis (FAB). Whereas deletion of either taI or taL causes a dramatic drop in myxovirescin production, deletion of both genes (DeltataIL) leads to the complete loss of myxovirescin production. These results suggest that both TaI and TaL PKSs might act in conjunction with a methyltransferase, reductases, and a monooxygenase to produce the 2-hydroxyvaleryl-S-ACP starter that is proposed to act as the biosynthetic primer in the initial condensation reaction with glycine. Polymerization of the remaining 11 acetates required for lactone formation is directed by 12 modules of Ta-1, TaO, and TaP megasynthetases. All modules, except for the first module of TaL, lack cognate acyltransferase (AT) domains. Furthermore, deletion of a discrete tandem AT-encoded by taV-blocks myxovirescin production; this suggests an "in trans" mode of action. To embellish the macrocycle with methyl and ethyl moieties, assembly of the myxovirescin scaffold is proposed to switch twice from PKS to 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA)-like biochemistry during biosynthesis. Disruption of the S-adenosylmethionine (SAM)-dependent methyltransferase, TaQ, shifts production toward two novel myxovirescin analogues, designated myxovirescin Q(a) and myxovirescin Q(c). NMR analysis of purified myxovirescin Q(a) revealed the loss of the methoxy carbon atom. This novel analogue lacks bioactivity against E. coli.
authors.affiliation: Pharmaceutical Biotechnology, Saarland University, Im Stadtwald, 66123 Saarbrücken, Germany.
authors.forename: Vesna
authors.initials: V
authors.lastname: Simunovic
authors.forename: Josef
authors.initials: J
authors.lastname: Zapp
authors.forename: Shwan
authors.initials: S
authors.lastname: Rachid
authors.forename: Daniel
authors.initials: D
authors.lastname: Krug
authors.forename: Peter
authors.initials: P
authors.lastname: Meiser
authors.forename: Rolf
authors.initials: R
authors.lastname: Müller
mesh_terms: D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000737:chemistry; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D017124:Conserved Sequence; D006904:Hydroxymethylglutaryl-CoA Synthase / Q000235:genetics / Q000378:metabolism*; D007783:Lactones / Q000737:chemistry / Q000378:metabolism; D008780:Methyltransferases / Q000378:metabolism; D008969:Molecular Sequence Data; D015394:Molecular Structure; D005810:Multigene Family / Q000235:genetics; D016941:Myxococcus xanthus / Q000737:chemistry / Q000378:metabolism; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism; D010453:Peptide Synthases / Q000235:genetics / Q000378:metabolism*; D048630:Polyketide Synthases / Q000235:genetics / Q000378:metabolism*; D012270:Ribosomes / Q000201:enzymology
chemical_list: D000900:Anti-Bacterial Agents; D007783:Lactones; D048630:Polyketide Synthases; C036969:myxovirescin A; D008780:Methyltransferases; D000217:Acyltransferases; D006904:Hydroxymethylglutaryl-CoA Synthase; D010453:Peptide Synthases; D010100:Oxygen