Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Provincial Engineering Technology Research Center of Seafood, Guangdong Province Engineering Laboratory for Marine Biological Products, Key Laboratory of Advanced Processing of Aquatic Product of Guangdong Higher Education Institution, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, China.
Collaborative Innovation Center of Seafood Deep Processing, Dalian Polytechnic University, Dalian 116034, China.
Molecules. 2022 Oct 12;27(20):6822. doi: 10.3390/molecules27206822.
Dissolved oxygen (DO) is an key factor for lipopeptide fermentation. To better understand the link between oxygen supply and lipopeptide productivity in CMT-6, the mechanism of DO on the synthesis of antimicrobial lipopeptides by CMT-6 was examined. The production of surfactin and iturin of CMT-6 was detected by liquid chromatography-mass spectrometer (LC-MS) under different DO conditions and transcriptome analysis was performed. At 100 and 200 rpm, the lipopeptides productions were 2753.62 mg/L and 3452.90 mg/L, respectively. There was no significant change in the yield of iturin but that of surfactin increased by 64.14%. Transcriptome analysis revealed that the enriched differential genes were concentrated in the GO term of oxidation-reduction process. The marked enrichment of the lipopeptides synthesis pathway, including microbial metabolism in diverse environments and carbon metabolism in the two-component system, were observed. More importantly, the expression levels of the four surfactin synthetase genes increased at higher DO, however, the iturin synthetase gene expression did not. Furthermore, modular surfactin synthetase was overexpressed (between 9- and 49-fold) at 200 rpm but not at 100 rpm, which is suggestive of efficient surfactin assembly resulting in surfactin overproduction. This study provides a theoretical basis for constructing engineering strains with high lipopeptide production to adapt to different DO.
溶解氧(DO)是脂肽发酵的关键因素。为了更好地理解供氧与 CMT-6 中脂肽生产力之间的联系,研究了 DO 对 CMT-6 合成抗菌脂肽的作用机制。通过液相色谱-质谱联用仪(LC-MS)在不同 DO 条件下检测到 CMT-6 产生的表面活性素和伊枯草菌素,同时进行了转录组分析。在 100 和 200 rpm 下,脂肽的产量分别为 2753.62 和 3452.90 mg/L。伊枯草菌素的产量没有显著变化,但表面活性素的产量增加了 64.14%。转录组分析表明,富集的差异基因集中在氧化还原过程的 GO 术语中。观察到脂肽合成途径的明显富集,包括微生物在各种环境中的代谢和双组分系统中的碳代谢。更重要的是,在较高 DO 下,四个表面活性素合成酶基因的表达水平增加,但伊枯草菌素合成酶基因的表达没有增加。此外,在 200 rpm 下,模块化表面活性素合成酶的表达(9 到 49 倍之间)增加,但在 100 rpm 下没有增加,这表明表面活性素的组装效率更高,导致表面活性素过量产生。这项研究为构建适应不同 DO 的高产脂肽工程菌株提供了理论基础。