Frontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Yaguan Road 135, Jinnan District, Tianjin, 300350, People's Republic of China.
SynBio Research Platform, Collaborative Innovation Centre of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Yaguan Road 135, Jinnan District, Tianjin, 300350, People's Republic of China.
J Ind Microbiol Biotechnol. 2020 Jul;47(6-7):551-562. doi: 10.1007/s10295-020-02275-7. Epub 2020 Jun 3.
Polymyxins are used as the last-line therapy against multidrug-resistant bacteria. However, their further clinical development needs to solve problems related to the presence of heterogeneous analogs, but there is still no platform or methods that can regulate the biosynthesis of polymyxin analogs. In this study, we present an approach to swap domains in the polymyxin gene cluster to regulate the production of different analogs. Following adenylation domain swapping, the proportion of polymyxin B1 increased from 41.36 to 52.90%, while that of B1-1 decreased from 18.25 to 3.09%. The ratio of polymyxin B1 and B3 following starter condensation domain swapping changed from 41.36 and 16.99 to 55.03 and 6.39%, respectively. The two domain-swapping strains produced 62.96% of polymyxin B1, 6.70% of B3 and 3.32% of B1-1. This study also revealed the presence of overflow fluxes between acetoin, 2,3-butanediol and polymyxin. To our best knowledge, this is the first report of engineering the polymyxin synthetase gene cluster in situ to regulate the relative proportions of polymyxin analogs. This research paves a way for regulating lipopeptide analogs and will facilitate the development of novel lipopeptide derivatives.
多黏菌素被用作治疗多重耐药菌的最后一线药物。然而，其进一步的临床开发需要解决与存在异质类似物相关的问题，但目前仍没有可以调节多黏菌素类似物生物合成的平台或方法。在本研究中，我们提出了一种在多黏菌素基因簇中交换结构域的方法来调节不同类似物的产生。在腺嘌呤酰化结构域交换后，多黏菌素 B1 的比例从 41.36%增加到 52.90%，而 B1-1 的比例从 18.25%降低至 3.09%。起始缩合结构域交换后，多黏菌素 B1 和 B3 的比例从 41.36%和 16.99%分别变为 55.03%和 6.39%。两株结构域交换菌株共产生了 62.96%的多黏菌素 B1、6.70%的 B3 和 3.32%的 B1-1。本研究还揭示了乙酰基-2,3-丁二醇和多黏菌素之间存在溢流通量。据我们所知，这是首次在原位工程化多黏菌素合成酶基因簇以调节多黏菌素类似物相对比例的报道。这项研究为调节脂肽类似物开辟了道路，并将有助于新型脂肽衍生物的开发。
