Identification of the novobiocin biosynthetic gene cluster of Streptomyces spheroides NCIB 11891
Identification of the novobiocin biosynthetic gene cluster of Streptomyces spheroides NCIB 11891
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DOI：
10.1128/aac.44.5.1214-1222.2000
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发表时间：
2000-05-01
影响因子：
4.9
通讯作者：
Heide, L
中科院分区：
文献类型：
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作者：
Steffensky, M;Mühlenweg, A;Heide, L
以同源脱氧核苷二磷酸(dNDP)-葡萄糖4,6-脱水酶基因片段为探针，克隆了球链霉菌NCIB 11891的新生物素生物合成基因簇。25.6 kb的双链测序显示存在23个假定的开放阅读框（orf），包括新生物素抗性基因gyrB(x)，以及至少11个可能在新生物素生物合成中起作用的orf。dndp -葡萄糖4,6-脱水酶片段的插入失活实验导致新生物素生产的消除，因为新生物素脱氧糖部分的生物合成被阻断。lividans TK24中新生素合成关键酶新生酸合成酶的异源表达进一步证实了所分析的基因参与了抗生素的生物合成。
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(x), 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.