Niu Guoqing, Li Lei, Wei Junhong, Tan Huarong
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
Chem Biol. 2013 Jan 24;20(1):34-44. doi: 10.1016/j.chembiol.2012.10.017.
Gougerotin, a peptidyl nucleoside antibiotic, possesses antitumor, antiviral, antibacterial, antimycoplasma, anthelmintic, and acaricidal activities. Here, we report the cloning of a complete gougerotin biosynthetic gene cluster from Streptomyces graminearus and heterologous production of gougerotin in Streptomyces coelicolor. Sequence analysis of a 28.7 kb DNA fragment indicated that the cluster consists of 25 open reading frames (ORFs). Gene disruption and genetic complementation experiments revealed that 15 of the 25 ORFs are required for gougerotin biosynthesis. A gougerotin biosynthetic pathway was proposed based on the analyses of bioinformatics and intermediates accumulated in selected gene inactivation mutants. These studies substantially promoted our understanding of gougerotin biosynthesis and provide "building blocks" for combinatorial biosynthesis using genes encoding different enzymes in nucleoside antibiotics.
谷氏菌素是一种肽基核苷类抗生素，具有抗肿瘤、抗病毒、抗菌、抗支原体、驱虫和杀螨活性。在此，我们报道了从禾本科链霉菌中克隆完整的谷氏菌素生物合成基因簇，并在天蓝色链霉菌中异源生产谷氏菌素。对一个28.7 kb DNA片段的序列分析表明，该基因簇由25个开放阅读框（ORF）组成。基因破坏和遗传互补实验表明，25个ORF中的15个是谷氏菌素生物合成所必需的。基于生物信息学分析和选定基因失活突变体中积累的中间体，提出了谷氏菌素生物合成途径。这些研究极大地促进了我们对谷氏菌素生物合成的理解，并为使用核苷类抗生素中编码不同酶的基因进行组合生物合成提供了“构件”。
