Key Laboratory of Carbohydrate Chemistry and Biotechnology of Ministry of Education, School of Life Sciences and Health Engineering, Jiangnan University, Wuxi 214122, PR China; School of Chemical and Material Engineering, Jiangnan University, Wuxi 214122, PR China.
Key Laboratory of Carbohydrate Chemistry and Biotechnology of Ministry of Education, School of Life Sciences and Health Engineering, Jiangnan University, Wuxi 214122, PR China; National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, School of Biotechnology, Jiangnan University, Wuxi 214122, PR China.
Bioresour Technol. 2024 Dec;414:131492. doi: 10.1016/j.biortech.2024.131492. Epub 2024 Sep 27.
Although Bacillus subtilis shows promise as a potential microbial cell factory for phospholipase D (PLD) expression, its production capacity remains insufficient. In this study, a secretory expression system, by co-optimization the promoter and signal peptides and employing a fed-batch fermentation strategy, was constructed to enhance expression of PLD from separate sources. The highest PLD production of 4056.9 U/mL was observed using this system, with a PLD production efficiency of 52.0 U/mL/h. Finally, a phosphatidic acid (PA) biosynthesis system was established using the constructed PLD as a catalyst, which achieved a PA yield of 219.1 g/L. This is the highest PLD production and PA yield reported globally to date. The protocol has significant potential for application for industrial PLD production as well as enzymatic phospholipids modification and also provides a valuable reference for overexpressing proteins in B. subtilis.
尽管枯草芽孢杆菌作为一种有潜力的微生物细胞工厂，可用于表达磷脂酶 D（PLD），但其生产能力仍然不足。在本研究中，构建了一种分泌表达系统，通过对启动子和信号肽进行协同优化，并采用分批补料发酵策略，以提高来自不同来源的 PLD 的表达量。使用该系统观察到最高的 PLD 产量为 4056.9 U/mL，PLD 生产效率为 52.0 U/mL/h。最后，使用构建的 PLD 作为催化剂建立了一个磷脂酸（PA）生物合成系统，实现了 219.1 g/L 的 PA 产量。这是迄今为止全球报道的最高的 PLD 产量和 PA 产量。该方案具有在工业上生产 PLD 以及酶法修饰磷脂的巨大应用潜力，也为枯草芽孢杆菌中过表达蛋白提供了有价值的参考。
