Identification and Manipulation of the Caprazamycin Gene Cluster Lead to New Simplified Liponucleoside Antibiotics and Give Insights into the Biosynthetic Pathway
Identification and Manipulation of the Caprazamycin Gene Cluster Lead to New Simplified Liponucleoside Antibiotics and Give Insights into the Biosynthetic Pathway
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DOI：
10.1074/jbc.m901258200
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发表时间：
2009-05-29
影响因子：
4.8
通讯作者：
Gust, Bertolt
中科院分区：
文献类型：
--
作者：
Kaysser, Leonard;Lutsch, Liane;Gust, Bertolt
卡普扎霉素是从链霉菌中分离到的一种有效的抗分枝杆菌脂核苷类抗生素。MK730-62F2，属于转移酶I抑制剂家族。它们的络合物结构是由5‘-(β-O-氨基)-甘草酸衍生而来，并含有一个独特的N-甲基二氮杂环。生物合成基因簇已经被鉴定、克隆和测序，这是转移酶I抑制剂的第一个基因簇。序列分析揭示了23个开放阅读框的存在，推测与卡普扎霉素的输出、抗性、调节和生物合成有关。基因簇在天蓝色链霉菌M512中的异源表达导致了非糖化生物活性卡普扎霉素衍生物的产生。一组基因缺失验证了cpz21簇的边界，cpz21的失活导致了新型简化的脂核苷抗生素的积累，这些抗生素缺乏3-甲基戊二酰基部分。因此，Cpz21被指定为卡普扎霉素生物合成中的一个酰基转移酶。在体内和在电子计算机中，对卡普扎霉素生物合成基因簇的分析可以首次提出生物合成途径，并为相关尿苷类抗生素的生物合成提供见解。
Caprazamycins are potent anti-mycobacterial liponucleoside antibiotics isolated from Streptomyces sp. MK730-62F2 and belong to the translocase I inhibitor family. Their complex structure is derived from 5'-(beta-O-aminoribosyl)-glycyluridine and comprises a unique N-methyldiazepanone ring. The biosynthetic gene cluster has been identified, cloned, and sequenced, representing the first gene cluster of a translocase I inhibitor. Sequence analysis revealed the presence of 23 open reading frames putatively involved in export, resistance, regulation, and biosynthesis of the caprazamycins. Heterologous expression of the gene cluster in Streptomyces coelicolor M512 led to the production of non-glycosylated bioactive caprazamycin derivatives. A set of gene deletions validated the boundaries of the cluster and inactivation of cpz21 resulted in the accumulation of novel simplified liponucleoside antibiotics that lack the 3-methylglutaryl moiety. Therefore, Cpz21 is assigned to act as an acyltransferase in caprazamycin biosynthesis. In vivo and in silico analysis of the caprazamycin biosynthetic gene cluster allows a first proposal of the biosynthetic pathway and provides insights into the biosynthesis of related uridyl-antibiotics.