title: Genetic manipulation revealing an unusual N-terminal region in a stand-alone non-ribosomal peptide synthetase involved in the biosynthesis of ramoplanins.
abstract: Ramoplanins produced by Actinoplanes are new structural class of lipopeptide and are currently in phase III clinical trials for the prevention of vancomycin-resistant enterococcal infections. The depsipeptide structures of ramoplanins are synthesized by non-ribosomal peptide synthetases (NRPS). Romo-orf17, a stand-alone NRPS, is responsible for the recruitment of Thr into the linear NRPS pathways for which the corresponding adenylation domain is absent. Here, systematical gene inactivation and complementation have been carried out in a Actinoplanes sp. using homologous recombination and site-specific integration methods. A hybrid gene coding for the N-terminal region of the stand-alone NRPS and the A-PCP domains of a heterologous NRPS restored production of ramoplanins. The results elucidate the unusual N-terminal region which is essential for the biosynthesis of ramoplanins.
authors.affiliation: State Key Lab of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, Shanghai, China.
authors.forename: Hai-Xue
authors.initials: HX
authors.lastname: Pan
authors.forename: Ji-An
authors.initials: JA
authors.lastname: Li
authors.forename: Lei
authors.initials: L
authors.lastname: Shao
authors.forename: Chun-Bao
authors.initials: CB
authors.lastname: Zhu
authors.forename: Jun-Sheng
authors.initials: JS
authors.lastname: Chen
authors.forename: Gong-Li
authors.initials: GL
authors.lastname: Tang
authors.forename: Dai-Jie
authors.initials: DJ
authors.lastname: Chen
mesh_terms: D000192:Actinomycetales / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular / Q000379:methods*; D019295:Computational Biology; D047630:Depsipeptides / Q000096:biosynthesis*; D005285:Fermentation; D017353:Gene Deletion; D005818:Genetic Engineering; D006023:Glycoproteins / Q000096:biosynthesis*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010453:Peptide Synthases / Q000235:genetics* / Q000378:metabolism
chemical_list: D047630:Depsipeptides; D006023:Glycoproteins; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase; C416073:ramoplanin A2