Minimizing endogenous cryptic plasmids to construct antibiotic-free expression systems for Escherichia coli Nissle 1917

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology Pub Date : 2024-01-25 DOI:10.1016/j.synbio.2024.01.006

Siyan Zhou , Linlin Zhao , Wenjie Zuo , Yilin Zheng , Ping Zhang , Yanan Sun , Yang Wang , Guocheng Du , Zhen Kang

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引用次数: 0

Abstract

The probiotic bacterium Escherichia coli 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 Bacteroides thetaiotaomicron 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.

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尽量减少内源性隐性质粒，构建大肠杆菌 Nissle 1917 的无抗生素表达系统

益生菌大肠埃希氏菌 Nissle 1917（EcN）在临床和生物工业中的应用前景十分广阔。然而，依赖抗生素来维持质粒携带的基因却掩盖了它的益处。在本研究中，我们通过对内源性隐性质粒 pMUT1 和 pMUT2 进行工程化处理来解决这一问题。去除非必要元素后，我们创造出了更稳定的衍生物 pMUT1NR△ 和 pMUT2HBC△。通过整合 sigma 因子上的结合基序，进一步构建了合成启动子，并将其用于表达 Bacteroides thetaiotaomicron 肝素酶 III 和外氨酸的生物合成。与传统的抗生素依赖型表达系统相比，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方面具有相当大的优势。

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来源期刊

Synthetic and Systems Biotechnology BIOTECHNOLOGY & APPLIED MICROBIOLOGY-

CiteScore

6.90

自引率

12.50%

发文量

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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