title: Production of the Ramoplanin Activity Analogue by Double Gene Inactivation.
abstract: Glycopeptides such as vancomycin and telavancin are essential for treating infections caused by Gram-positive bacteria. But the dwindling availability of new antibiotics and the emergence of resistant bacteria are making effective antibiotic treatment increasingly difficult. Ramoplanin, an inhibitor of bacterial cell wall biosynthesis, is a highly effective antibiotic against a wide range of Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus, vancomycin-intermediate resistant Clostridium difficile and vancomycin-resistant Enterococcus sp. Here, two tailoring enzyme genes in the biosynthesis of ramoplanin were deleted by double in-frame gene knockouts to produce new ramoplanin derivatives. The deschlororamoplanin A2 aglycone was purified and its structure was identified with LC-MS/MS. Deschlororamoplanin A2 aglycone and ramoplanin aglycone showed similar activity to ramoplanin A2. The results showed that α-1,2-dimannosyl disaccharide at Hpg11 and chlorination at Chp17 in the ramoplanin structure are not essential for its antimicrobial activity. This work provides new precursor compounds for the semisynthetic modification of ramoplanin.
authors.affiliation: State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, 1320 West Beijing Rd., Shanghai 200040, China.
authors.forename: Jungang
authors.initials: J
authors.lastname: Han
authors.affiliation: State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, 1320 West Beijing Rd., Shanghai 200040, China.|School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Rd., Minhang District, Shanghai 200240, China.
authors.forename: Junsheng
authors.initials: J
authors.lastname: Chen
authors.affiliation: State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, 1320 West Beijing Rd., Shanghai 200040, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Shao
authors.affiliation: State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, 1320 West Beijing Rd., Shanghai 200040, China.
authors.forename: Junliang
authors.initials: J
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, 1320 West Beijing Rd., Shanghai 200040, China.
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Dong
authors.affiliation: State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, 1320 West Beijing Rd., Shanghai 200040, China.
authors.forename: Pengyu
authors.initials: P
authors.lastname: Liu
authors.affiliation: State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, 1320 West Beijing Rd., Shanghai 200040, China.
authors.forename: Daijie
authors.initials: D
authors.lastname: Chen
mesh_terms: D000192:Actinomycetales / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D047630:Depsipeptides / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D013293:Enterococcus faecalis / Q000187:drug effects; D005285:Fermentation; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D006023:Glycoproteins / Q000378:metabolism / Q000494:pharmacology; D055624:Methicillin-Resistant Staphylococcus aureus / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D016133:Polymerase Chain Reaction; D020713:Vancomycin Resistance
chemical_list: D000900:Anti-Bacterial Agents; D047630:Depsipeptides; D006023:Glycoproteins; C472958:ramoplanose aglycon; C041438:ramoplanin; C416073:ramoplanin A2