Identification of the Novobiocin Biosynthetic Gene Cluster of Streptomyces spheroides NCIB 11891

作者：Marion Steffensky, Agnes Mühlenweg, Zhaoxin Wang, Shu-Ming Li, Lutz E. Heide · 发表于：Antimicrobial Agents and Chemotherapy · 年份：2000 · DOI：10.1128/aac.44.5.1214-1222.2000 · 被引用次数：196 · 研究领域：Microbial Natural Products and Biosynthesis、Enzyme Structure and Function、Biochemical and Molecular Research

The novobiocin biosynthetic gene cluster from Streptomyces spheroides NCIB 11891 was cloned by using homologous deoxynucleoside diphosphate (dNDP)-glucose 4,6-dehydratase gene fragments as probes. Double-stranded sequencing of 25.6 kb revealed the presence of 23 putative open reading frames (ORFs), including the gene for novobiocin resistance, gyrB(r), and at least 11 further ORFs to which a possible role in novobiocin biosynthesis could be assigned. An insertional inactivation experiment with a dNDP-glucose 4, 6-dehydratase fragment resulted in abolishment of novobiocin production, since biosynthesis of the deoxysugar moiety of novobiocin was blocked. Heterologous expression of a key enzyme of novobiocin biosynthesis, i.e., novobiocic acid synthetase, in Streptomyces lividans TK24 further confirmed the involvement of the analyzed genes in the biosynthesis of the antibiotic.