title: Investigation of halogenation during the biosynthesis of ramoplanin in Actinoplanes sp. ATCC33076.
abstract: Ramoplanin and enduracidin are lipopeptide antibiotics effective against Gram-positive pathogens, which share close similarity in structure and biosynthetic pathway. Both compounds have chlorine atoms attached to 4-hydroxyphenylglycine (Hpg) but with different chlorinating sites and levels. Here, to probe the factor affecting the site and level of halogenation, gene inactivation and heterologous expression were carried out in Actinoplanes sp. ATCC33076 by homologous recombination. Metabolite analysis confirmed that ram20 encodes the only halogenase in ramoplanin biosynthetic pathway, and enduracidin halogenase End30 could heterologously complement the ram20-deficient mutant. Additionally, the mannosyltransferase-deficient mutant produces a dichlorinated ramoplanin aglycone with the halogenation site at Hpg(13). This study has refined our understanding of how halogenation occurs in ramoplanin biosynthetic pathway, and lays the foundation for further exploitation of ramoplanin and enduracidin halogenase in combinatorial biosynthesis.
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Rd., Minhang District, Shanghai, 200240, China.|State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, 1320 West Beijing Rd., Shanghai, 200040, China.|Engineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, SJTU, 800 Dongchuan Rd., Minhang District, Shanghai, 200240, China.
authors.forename: Jun-Sheng
authors.initials: JS
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: Min
authors.initials: M
authors.lastname: Su
authors.affiliation: State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, 1320 West Beijing Rd., Shanghai, 200040, China. shaolei00@gmail.com.
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: Yuan-Xi
authors.initials: YX
authors.lastname: Wang
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: Hui-Min
authors.initials: HM
authors.lastname: Lin
authors.affiliation: State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, 1320 West Beijing Rd., Shanghai, 200040, China. chen_daijie@yahoo.com.
authors.forename: Dai-Jie
authors.initials: DJ
authors.lastname: Chen
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D002713:Chlorine / Q000378:metabolism*; D047630:Depsipeptides / Q000096:biosynthesis*; D015870:Gene Expression; D055786:Gene Knockout Techniques; D005816:Genetic Complementation Test; D054879:Halogenation; D059765:Homologous Recombination; D008848:Micromonosporaceae / Q000378:metabolism*; D010456:Peptides, Cyclic / Q000096:biosynthesis
chemical_list: D000900:Anti-Bacterial Agents; D047630:Depsipeptides; D010456:Peptides, Cyclic; C505063:enduracidin A; C041438:ramoplanin; D002713:Chlorine
keywords: Antibiotics; Biosynthesis; Enduracidin; Halogenase; Ramoplanin