title: Functional identification of the gene encoding the enzyme involved in mannosylation in ramoplanin biosynthesis in Actinoplanes sp.
abstract: Ramoplanin is a lipopeptide antibiotic active against multi-drug-resistant, Gram-positive pathogens. Structurally, it contains a di-mannose moiety attached to the peptide core at Hpg(11). The biosynthetic gene cluster of ramoplanin has already been reported and the assembly of the depsipeptide has been elucidated but the mechanism of transferring sugar moiety to the peptide core remains unclear. Sequence analysis of the biosynthetic gene cluster indicated ramo-orf29 was a mannosyltransferase candidate. To investigate the involvement of ramo-orf29 in ramoplanin biosynthesis, gene inactivation and complementation have been conducted in Actinoplanes sp. ATCC 33076 by homologous recombination. Metabolite analysis revealed that the ramo-orf29 inactivated mutant produced no ramoplanin but the ramoplanin aglycone. Thus, ramo-orf29 codes for the mannosyltransferase in the ramoplanin biosynthesis pathway. This lays the foundation for further exploitation of the ramoplanin mannosyltransferase and aglycone in combinatorial biosynthesis.
authors.affiliation: State Key Lab of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, 1320 West Beijing Road, Shanghai, 200040, China.
authors.forename: Jun-Sheng
authors.initials: JS
authors.lastname: Chen
authors.forename: Yuan-Xi
authors.initials: YX
authors.lastname: Wang
authors.forename: Lei
authors.initials: L
authors.lastname: Shao
authors.forename: Hai-Xue
authors.initials: HX
authors.lastname: Pan
authors.forename: Ji-An
authors.initials: JA
authors.lastname: Li
authors.forename: Hui-Min
authors.initials: HM
authors.lastname: Lin
authors.forename: Xiao-Jing
authors.initials: XJ
authors.lastname: Dong
authors.forename: Dai-Jie
authors.initials: DJ
authors.lastname: Chen
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D047630:Depsipeptides / Q000096:biosynthesis*; D055786:Gene Knockout Techniques; D005816:Genetic Complementation Test; D059765:Homologous Recombination; D008358:Mannose / Q000378:metabolism*; D008364:Mannosyltransferases / Q000235:genetics* / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D008848:Micromonosporaceae / Q000201:enzymology* / Q000235:genetics*; D016366:Open Reading Frames; D017422:Sequence Analysis, DNA; D013211:Staphylococcus aureus / Q000187:drug effects
chemical_list: D000900:Anti-Bacterial Agents; D047630:Depsipeptides; C041438:ramoplanin; D008364:Mannosyltransferases; D008358:Mannose