Zhou Siyan, Zhao Linlin, Zuo Wenjie, Zheng Yilin, Zhang Ping, Sun Yanan, Wang Yang, Du Guocheng, Kang Zhen
The Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China.
The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China.
Synth Syst Biotechnol. 2024 Jan 25;9(1):165-175. doi: 10.1016/j.synbio.2024.01.006. eCollection 2024 Mar.
The probiotic bacterium Nissle 1917 (EcN) holds significant promise for use in clinical and biological industries. However, the reliance on antibiotics to maintain plasmid-borne genes has overshadowed its benefits. In this study, we addressed this issue by engineering the endogenous cryptic plasmids pMUT1 and pMUT2. The non-essential elements were removed to create more stable derivatives pMUT1NR△ and pMUT2HBC△. Synthetic promoters by integrating binding motifs on sigma factors were further constructed and applied for expression of heparinase III and the biosynthesis of ectoine. Compared to traditional antibiotic-dependent expression systems, our newly constructed antibiotic-free expression systems offer considerable advantages for clinical and synthetic biology applications.
益生菌菌株Nissle 1917（EcN）在临床和生物产业中具有巨大的应用潜力。然而，依赖抗生素来维持质粒携带的基因掩盖了它的优势。在本研究中，我们通过对内生隐蔽质粒pMUT1和pMUT2进行工程改造来解决这个问题。去除了非必需元件以创建更稳定的衍生物pMUT1NR△和pMUT2HBC△。通过整合sigma因子上的结合基序构建了合成启动子，并将其应用于肝素酶III的表达和四氢嘧啶的生物合成。与传统的依赖抗生素的表达系统相比，我们新构建的无抗生素表达系统在临床和合成生物学应用中具有显著优势。
