Cloning of the staurosporine biosynthetic gene cluster from Streptomyces sp TP-A0274 and its heterologous expression in Streptomyces lividans
Cloning of the staurosporine biosynthetic gene cluster from Streptomyces sp TP-A0274 and its heterologous expression in Streptomyces lividans
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DOI：
10.7164/antibiotics.55.1063
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发表时间：
2002-12-01
影响因子：
3.3
通讯作者：
Furumai, T
中科院分区：
文献类型：
--
作者：
Onaka, H;Taniguchi, S;Furumai, T
星形孢菌素是吲哚咔唑类抗生素的代表性成员。从链霉菌TP-A0274中克隆了跨度为20-kb的整个星形孢菌素生物合成和调控基因簇，并进行了测序。该基因簇由14个ORF组成，氨基酸序列同源性搜索显示，它包含三个基因，staO，staD和staP，编码参与吲哚并咔唑糖苷配基生物合成的酶，两个基因，staG和staN，用于糖苷配基和脱氧糖之间的键形成，八个基因，staA，staB，staE，staJ，staI，staK，staMA和staMB，对于脱氧糖的生物合成和一个基因，星星是一个转录调节因子。将含有完整的星形孢菌素生物合成基因的38-kb片段克隆到pTOYAMAcos中进行异源基因表达，证实了其在星形孢菌素生物合成中的作用。此外，色吡咯酸合成酶的基因的分布，吲哚咔唑苷元的生物合成的关键酶，放线菌进行了调查，rebD同源物只存在于菌株产生吲哚咔唑抗生素。
Staurosporine is a representative member of indolocarbazole antibiotics. The entire staurosporine biosynthetic and regulatory gene cluster spanning 20-kb was cloned from Streptomyces sp. TP-A0274 and sequenced.. The gene cluster consists of 14 ORFs and the amino acid sequence homology search revealed that it contains three genes, staO, staD, and staP, coding for the enzymes involved in the indolocarbazole aglycone biosynthesis, two genes, staG and staN, for the bond formation between the aglycone and deoxysugar, eight genes, staA, staB, staE, staJ, staI, staK, staMA, and staMB, for the deoxysugar biosynthesis and one gene, staR is a transcriptional regulator. Heterologous gene expression of a 38-kb fragment containing a complete set of the biosynthetic genes for staurosporine cloned into pTOYAMAcos confirmed its role in staurosporine biosynthesis. Moreover, the distribution of the gene for chromopyrrolic acid synthase, the key enzyme for the biosynthesis of indolocarbazole aglycone, in actinomycetes was investigated, and rebD homologs were shown to exist only in the strains producing indolocarbazole antibiotics.