Achieving robust synthetic tolerance in industrial E. coli through negative auto-regulation of a DsrA-Hfq module

IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Synthetic and Systems Biotechnology Pub Date : 2024-04-11 DOI:10.1016/j.synbio.2024.04.003
Xiaofeng Yang , Jingduan Yang , Haozheng Huang , Xiaofang Yan , Xiaofan Li , Zhanglin Lin
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Abstract

In industrial fermentation processes, microorganisms often encounter acid stress, which significantly impact their productivity. This study focused on the acid-resistant module composed of small RNA (sRNA) DsrA and the sRNA chaperone Hfq. Our previous study had shown that this module improved the cell growth of Escherichia coli MG1655 at low pH, but failed to obtain this desired phenotype in industrial strains. Here, we performed a quantitative analysis of DsrA-Hfq module to determine the optimal expression mode. We then assessed the potential of the CymR-based negative auto-regulation (NAR) circuit for industrial application, under different media, strains and pH levels. Growth assay at pH 4.5 revealed that NAR-05D04H circuit was the best acid-resistant circuit to improve the cell growth of E. coli MG1655. This circuit was robust and worked well in the industrial lysine-producing strain E. coli SCEcL3 at a starting pH of 6.8 and without pH control, resulting in a 250 % increase in lysine titer and comparable biomass in shaking flask fermentation compared to the parent strain. This study showed the practical application of NAR circuit in regulating DsrA-Hfq module, effectively and robustly improving the acid tolerance of industrial strains, which provides a new approach for breeding industrial strains with tolerance phenotype.

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通过 DsrA-Hfq 模块的负自动调节实现工业大肠杆菌的稳健合成耐受性
在工业发酵过程中,微生物经常会遇到酸胁迫,从而严重影响其生产率。这项研究的重点是由小核糖核酸(sRNA)DsrA和sRNA伴侣蛋白Hfq组成的耐酸模块。我们之前的研究表明,该模块改善了大肠杆菌 MG1655 在低 pH 值条件下的细胞生长,但在工业菌株中却未能获得这种理想的表型。在此,我们对 DsrA-Hfq 模块进行了定量分析,以确定最佳表达模式。然后,我们评估了基于 CymR 的负自动调节(NAR)电路在不同培养基、菌株和 pH 值条件下的工业应用潜力。在 pH 值为 4.5 的条件下进行的生长试验表明,NAR-05D04H 电路是改善大肠杆菌 MG1655 细胞生长的最佳耐酸电路。在起始 pH 值为 6.8 且无 pH 值控制的情况下,该电路在工业赖氨酸生产菌株大肠杆菌 SCEcL3 中运行良好,与母菌株相比,在摇瓶发酵中赖氨酸滴度提高了 250%,生物量也相当可观。这项研究表明,NAR回路在调控DsrA-Hfq模块方面的实际应用,能有效、稳健地提高工业菌株的耐酸性,为培育具有耐酸表型的工业菌株提供了一种新方法。
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来源期刊
Synthetic and Systems Biotechnology
Synthetic and Systems Biotechnology BIOTECHNOLOGY & APPLIED MICROBIOLOGY-
CiteScore
6.90
自引率
12.50%
发文量
90
审稿时长
67 days
期刊介绍: Synthetic and Systems Biotechnology aims to promote the communication of original research in synthetic and systems biology, with strong emphasis on applications towards biotechnology. This journal is a quarterly peer-reviewed journal led by Editor-in-Chief Lixin Zhang. The journal publishes high-quality research; focusing on integrative approaches to enable the understanding and design of biological systems, and research to develop the application of systems and synthetic biology to natural systems. This journal will publish Articles, Short notes, Methods, Mini Reviews, Commentary and Conference reviews.
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