title: Inactivation and identification of three genes encoding glycosyltransferase required for biosynthesis of nogalamycin.
abstract: Nogalamycin is an anthracycline antitumor antibiotic, consisting of the aromatic aglycone attached with a nogalose and a nogalamine. At present, the biosynthesis pathway of nogalamycin, especially the glycosylation mechanism of the two deoxysugar moieties, had still not been extensively investigated in vivo. In this study, we inactivated the three glycotransferase genes in the nogalamycin-produced strain, and investigated the function of these genes by analyzing the metabolites profiles in the fermentation broth. The in-frame deletion of snogD and disruption of snogE abolished the production of nogalamycin completely, indicating that the gene products of snogD and snogE are essential to the biosynthesis of nogalamycin. On the other hand, in-frame deletion of snogZ does not abolish the production of nogalamycin, but production yield was reduced to 28% of the wild type, implying that snogZ gene may involved in the activation of other glycotransferases in nogalamycin biosynthesis. This study laid the foundation of modification of nogalamycin biosynthesis/production by genetic engineering methods.
authors.affiliation: State Key Lab of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, Shanghai, People's Republic of China.
authors.forename: Lei
authors.initials: L
authors.lastname: Shao
authors.affiliation: State Key Lab of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, Shanghai, People's Republic of China.
authors.forename: Xuanwen
authors.initials: X
authors.lastname: Shi
authors.affiliation: State Key Lab of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, Shanghai, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.affiliation: State Key Lab of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, Shanghai, People's Republic of China.
authors.forename: Xiaorong
authors.initials: X
authors.lastname: Gao
authors.affiliation: State Key Lab of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, Shanghai, People's Republic of China.
authors.forename: Tian
authors.initials: T
authors.lastname: Pu
authors.affiliation: Department of Traditional Chinese Medicine, Agronomy College of Henan Agricultural University, Zhengzhou, People's Republic of China.
authors.forename: Bingji
authors.initials: B
authors.lastname: Ma
authors.affiliation: College of Life Science, Nanjing, People's Republic of China.
authors.forename: Siyuan
authors.initials: S
authors.lastname: Wang
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D020868:Gene Silencing; D005818:Genetic Engineering / Q000379:methods*; D016695:Glycosyltransferases / Q000172:deficiency* / Q000235:genetics* / Q000378:metabolism; D009621:Nogalamycin / Q000096:biosynthesis*; D016415:Sequence Alignment; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism
chemical_list: D016695:Glycosyltransferases; D009621:Nogalamycin
keywords: biosynthesis; gene disruption; glycosyltransferase (GTs); homologous recombination; nogalamycin