期刊
MICROBIOLOGY-UK
卷 146, 期 -, 页码 611-619出版社
SOC GENERAL MICROBIOLOGY
DOI: 10.1099/00221287-146-3-611
关键词
Streptomyces noursei; gene cluster; macrolide antibiotic
类别
Streptomyces noursei ATCC 11455 produces the antifungal polyene antibiotic nystatin containing the deoxysugar moiety mycosamine. Part of the deoxythymidyl diphosphate (TDP)-glucose dehydratase gene (gdhA) known to be involved in deoxysugar biosynthesis was amplified by PCR from genomic DNA of S, noursei ATCC 11455, A gene library for S, noursei was made and screened with the gdhA probe. Several overlapping phage clones covering about 30 kb of the S, noursei genome were physically mapped, A partial DNA sequencing analysis of this region resulted in the identification of several putative genes typical of macrolide antibiotic biosynthetic gene clusters, A gene-transfer system for S, noursei has been established, and gene deletion or disruption experiments within the putative biosynthetic gene cluster were performed, All of the knock-out mutants retained the ability to produce nystatin, suggesting that the identified gene cluster is not involved in biosynthesis of this antibiotic. Culture extracts from the wild-type strain and three knock-out mutants were analysed by TLC followed by a bioassay against Micrococcus luteus. Two antibacterial compounds were found to be synthesized by the wild-type strain while only one was produced by the mutants, This provided evidence for the involvement of the identified gene cluster in the biosynthesis of a presumably novel antibacterial macrolide antibiotic in S, noursei.
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