State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, China.
Shandong Provincial Key Laboratory of Synthetic Biology, CAS Key Laboratory of Biofuels at Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266101, China.
Mar Drugs. 2021 Jan 11;19(1):29. doi: 10.3390/md19010029.
Bafilomycin A is the representative compound of the plecomacrolide natural product family. This 16-membered ring plecomacrolide has potent antifungal and vacuolar H-ATPase inhibitory activities. In our previous work, we identified a bafilomycin biosynthetic gene cluster () from the marine bacterium ATCC BAA-1276, wherein a family regulatory gene and an family regulatory gene were revealed based on bioinformatics analysis. In this study, the positive regulatory roles of and for bafilomycin biosynthesis are characterized through gene inactivation and overexpression. Compared to the wild-type strain, the knockout of either or completely abolished the production of bafilomycins. The overexpression of or led to 1.3- and 0.5-fold increased production of bafilomycins, respectively. A genetically engineered strain (SLO-08) with overexpression and inactivation of the biosynthetic genes and , solely produced bafilomycin A with the titer of 535.1 ± 25.0 mg/L in an optimized fermentation medium in shaking flasks. This recombinant strain holds considerable application potential in large-scale production of bafilomycin A for new drug development.
巴弗洛霉素 A 是聚醚类天然产物家族的代表性化合物。这种 16 元环聚醚类化合物具有很强的抗真菌和液泡 H+-ATP 酶抑制活性。在我们之前的工作中，我们从海洋细菌 ATCC BAA-1276 中鉴定出一个巴弗洛霉素生物合成基因簇 ()，通过生物信息学分析揭示了一个 家族调控基因 和一个 家族调控基因 。在这项研究中，通过基因失活和过表达来表征 和 对巴弗洛霉素生物合成的正调控作用。与野生型 菌株相比，敲除 或 完全消除了巴弗洛霉素的产生。过表达 或 分别导致巴弗洛霉素产量增加了 1.3 倍和 0.5 倍。通过过表达 和失活生物合成基因 和 ，构建了一株遗传工程 菌株 (SLO-08)，该菌株在优化的摇瓶发酵培养基中仅产生巴弗洛霉素 A，产量为 535.1 ± 25.0 mg/L。该重组菌株在大规模生产巴弗洛霉素 A 用于新药开发方面具有很大的应用潜力。
