Identification and characterization of anaerobically activated promoters in Escherichia coli

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology Pub Date : 2025-03-04 DOI:10.1016/j.jbiotec.2025.03.002

Sen Yang , Chao-Hao Guo , Wen-Yue Tong , Xiao-Yun Liu , Jing-Chen Li , Ming Kang

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引用次数: 0

Abstract

Anaerobically activated promoters in Escherichia coli play crucial roles in transcriptional regulation during cellular responses to decreased oxygen concentrations and serve as essential tools for implementing dynamic regulation in metabolic engineering. These promoters exhibit transcriptional activity only under low-oxygen or anaerobic conditions. To discover novel anaerobically activated promoters, this study selected 11 native promoters from E. coli databases and characterized their activities using flow cytometry. Subsequently, we optimized the key elements of these promoters and re-evaluated their activities to investigate the impact of functional elements on promoter performance. Furthermore, we verified the regulatory mechanisms of these promoters by knocking out host regulatory genes. Finally, we characterized the promoters' responsiveness to aerobic-anaerobic transitions by rapidly switching cultivation environments during host growth. This study identified several novel anaerobically activated promoters and comprehensively characterized their performance and features from multiple aspects. The identified promoters provide new tools for oxygen-limited or anaerobic production in metabolic engineering, while the findings from promoter element optimization offer valuable references for the design of anaerobically activated promoters.

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大肠杆菌厌氧激活启动子的鉴定与表征。

大肠杆菌的厌氧激活启动子在细胞对低氧浓度的转录调控中起着至关重要的作用，是代谢工程中实现动态调控的重要工具。这些启动子仅在低氧或厌氧条件下表现出转录活性。为了发现新的厌氧激活启动子，本研究从大肠杆菌数据库中选择了11个天然启动子，并利用流式细胞术对其活性进行了表征。随后，我们优化了这些启动子的关键元素，并重新评估了它们的活性，以研究功能元素对启动子性能的影响。此外，我们通过敲除宿主调控基因验证了这些启动子的调控机制。最后，我们通过在宿主生长过程中快速切换培养环境来表征启动子对好氧-厌氧转变的响应性。本研究鉴定了几种新型的厌氧激活启动子，并从多个方面对其性能和特点进行了综合表征。所鉴定的启动子为代谢工程中的限氧或厌氧生产提供了新的工具，而启动子元件优化的研究结果为厌氧激活启动子的设计提供了有价值的参考。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of biotechnology 工程技术-生物工程与应用微生物

CiteScore

8.90

自引率

2.40%

发文量

190

审稿时长

45 days

期刊介绍： The Journal of Biotechnology has an open access mirror journal, the Journal of Biotechnology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. The Journal provides a medium for the rapid publication of both full-length articles and short communications on novel and innovative aspects of biotechnology. The Journal will accept papers ranging from genetic or molecular biological positions to those covering biochemical, chemical or bioprocess engineering aspects as well as computer application of new software concepts, provided that in each case the material is directly relevant to biotechnological systems. Papers presenting information of a multidisciplinary nature that would not be suitable for publication in a journal devoted to a single discipline, are particularly welcome.