文献类型：期刊文献

英文题名：Metabolic engineering of Bacillus subtilis for high-yield surfactin production

作者：Wang, Yong[1];Li, Hanqing[1];Mao, Yue[1];Li, Pengfei[1,2];Wu, Haizhen[1,2];Ye, Jiang[1];Zhang, Huizhan[1]

机构：[1]East China Univ Sci & Technol, Sch Biotechnol, Dept Appl Biol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Fermentat Engn Expt Teaching Demonstrat Ctr, Shanghai 200237, Peoples R China

年份：2026

卷号：19

期号：1

外文期刊名：BIOTECHNOLOGY FOR BIOFUELS AND BIOPRODUCTS

收录：;EI(收录号：20262020725795);WOS:【SCI-EXPANDED(收录号:WOS:001765848600001)】；

基金：This work was supported by the National Natural Science Foundation of China, 41530318.

语种：英文

外文关键词：Surfactin; Bacillus subtilis; Metabolic engineering; P43 promoter; Tolerance engineering

摘要：The industrial application of surfactin has been constrained by the low production capacity of Bacillus subtilis. This study developed a high-yield, high-tolerance surfactin-producing strain through a multi-level metabolic engineering strategy. First, we constructed an enhanced P43 promoter (P43LN) by extending its upstream region from 260 to 520 bp and pairing it with a cognate 5 ' UTR, which exhibited more than threefold higher activity than the commonly used core P43 promoter, providing a versatile tool for stable and high-level expression of key genes. Based on this, the native srfA promoter in the oilfield-derived B. subtilis TD7 was replaced with the P43 promoter, yielding the engineered strain TP1. Subsequently, systematic knockout of competing antimicrobial peptide synthesis gene clusters (pps, bac, and pks) increased surfactin titer to 2.78 g/L. Further synergistic enhancement of NADPH regeneration (via zwf overexpression) and cellular tolerance (via efflux pump gene yerP overexpression) resulted in the final strain TS8, which achieved a shake-flask surfactin titer of 3.51 g/L, representing a 102.7% increase in per-OD600 productivity. This work successfully integrates promoter engineering, metabolic flux redistribution, and tolerance modules, not only significantly improving surfactin production but also providing a powerful expression and systematic optimization toolkit for B. subtilis, laying a technical foundation for the efficient biomanufacturing of complex natural products.