Enhanced production of validamycin A in Streptomyces hygroscopicus 5008 by engineering validamycin biosynthetic gene cluster
Enhanced production of validamycin A in Streptomyces hygroscopicus 5008 by engineering validamycin biosynthetic gene cluster
复制标题
DOI：
10.1007/s00253-014-5943-9
复制
发表时间：
2014-07
影响因子：
5
通讯作者：
Tan-Che Zhou;Byung-gee Kim;J. Zhong
中科院分区：
文献类型：
--
作者：
Tan-Che Zhou;Byung-gee Kim;J. Zhong
井冈霉素 A (VAL-A) 是一种广泛使用的抗真菌抗生素，用于治疗水稻和其他植物的纹枯病。它可由农工业副产品由吸水链霉菌5008生产。为了提高其生产滴度，在这项工作中，整个val基因簇在S中串联扩增。 hygrooscopyus5008通过将zouA介导的DNA扩增系统整合到valgene簇的两个边界之间，产生valgene簇的多个拷贝（主要是3到5个）。通过Southern印迹和发酵实验证实了扩增拷贝的遗传稳定性。在摇瓶发酵中，与野生型菌株相比，重组菌株 (TC03) 的 VAL-A 生产滴度提高了 34%，而 TC03 中中间体有效木胺 A 的积累减少。此外，TC03 中的结构基因转录水平和 ValG 酶活性均显着增加。这项工作表明，valgene 簇的扩增是提高 S 的 VAL-A 产量的有效策略。 hygrooscopyus5008，所获得的信息将有助于通过基因簇扩增工程设计其他有趣的抗生素生物合成。
Validamycin A (VAL-A) is a widely used antifungal antibiotic for the treatment of sheath blight disease of rice and other plants. It can be produced from agro-industrial by-products byStreptomyces hygroscopicus5008. To enhance its production titer, in this work, the entirevalgene cluster was amplified in tandem inS. hygroscopicus5008 by integrating the zouA-mediated DNA amplification system into between the two boundaries ofvalgene cluster, resulting in multiple copies (mainly three to five) of thevalgene cluster. The genetic stability of the amplified copies was confirmed by Southern blot and fermentation experiments. In shake flask fermentation, the recombinant strain (TC03) led to a 34 % enhancement of VAL-A production titer compared to that of the wild-type strain, while the accumulation of intermediate validoxylamine A was decreased in TC03. Additionally, both the structural gene transcription levels and the ValG enzyme activity were significantly increased in TC03. This work demonstrated that the amplification of thevalgene cluster was an efficient strategy to enhance VAL-A production byS. hygroscopicus5008, and the information obtained would be helpful for engineering other interesting antibiotic biosynthesis by gene cluster amplification.