里氏链霉菌DSM 40489香豆霉素A（1）生物合成基因簇的鉴定
Identification of the coumermycin A(1) biosynthetic gene cluster of Streptomyces rishiriensis DSM 40489.
作者信息
Wang Z X, Li S M, Heide L
机构信息
Pharmazeutische Biologie, Pharmazeutisches Institut, Eberhard-Karls-Universität Tübingen, 72076 Tübingen, Germany.
出版信息
Antimicrob Agents Chemother. 2000 Nov;44(11):3040-8. doi: 10.1128/AAC.44.11.3040-3048.2000.
Abstract
The biosynthetic gene cluster of the aminocoumarin antibiotic coumermycin A(1) was cloned by screening of a cosmid library of Streptomyces rishiriensis DSM 40489 with heterologous probes from a dTDP-glucose 4,6-dehydratase gene, involved in deoxysugar biosynthesis, and from the aminocoumarin resistance gyrase gene gyrB(r). Sequence analysis of a 30.8-kb region upstream of gyrB(r) revealed the presence of 28 complete open reading frames (ORFs). Fifteen of the identified ORFs showed, on average, 84% identity to corresponding ORFs in the biosynthetic gene cluster of novobiocin, another aminocoumarin antibiotic. Possible functions of 17 ORFs in the biosynthesis of coumermycin A(1) could be assigned by comparison with sequences in GenBank. Experimental proof for the function of the identified gene cluster was provided by an insertional gene inactivation experiment, which resulted in an abolishment of coumermycin A(1) production.
摘要
通过用来自参与脱氧糖生物合成的dTDP - 葡萄糖4,6 - 脱水酶基因以及氨基香豆素抗性gyrase基因gyrB(r)的异源探针筛选里氏链霉菌DSM 40489的黏粒文库，克隆了氨基香豆素抗生素香豆霉素A(1)的生物合成基因簇。对gyrB(r)上游30.8 kb区域的序列分析揭示了28个完整的开放阅读框（ORF）的存在。在已鉴定的ORF中，有15个与另一种氨基香豆素抗生素新生霉素生物合成基因簇中的相应ORF平均具有84%的同一性。通过与GenBank中的序列比较，可以确定17个ORF在香豆霉素A(1)生物合成中的可能功能。插入基因失活实验为所鉴定基因簇的功能提供了实验证据，该实验导致香豆霉素A(1)的产生被消除。
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