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基于群体感应系统的肺炎克雷伯氏菌代谢通量调控。

The metabolic flux regulation of Klebsiella pneumoniae based on quorum sensing system.

机构信息

College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, PR China.

Centre for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao 266580, PR China.

出版信息

Sci Rep. 2016 Dec 7;6:38725. doi: 10.1038/srep38725.

Abstract

Quorum-sensing (QS) systems exist universally in bacteria to regulate multiple biological functions. Klebsiella pneumoniae, an industrially important bacterium that produces bio-based chemicals such as 2,3-butanediol and acetoin, can secrete a furanosyl borate diester (AI-2) as the signalling molecule mediating a QS system, which plays a key regulatory role in the biosynthesis of secondary metabolites. In this study, the molecular regulation and metabolic functions of a QS system in K. pneumoniae were investigated. The results showed that after the disruption of AI-2-mediated QS by the knockout of luxS, the production of acetoin, ethanol and acetic acid were relatively lower in the K. pneumoniae mutant than in the wild type bacteria. However, 2,3-butanediol production was increased by 23.8% and reached 54.93 g/L. The observed enhancement may be attributed to the improvement of the catalytic activity of 2,3-butanediol dehydrogenase (BDH) in transforming acetoin to 2,3-butanediol. This possibility is consistent with the RT-PCR-verified increase in the transcriptional level of budC, which encodes BDH. These results also demonstrated that the physiological metabolism of K. pneumoniae was adversely affected by a QS system. This effect was reversed through the addition of synthetic AI-2. This study provides the basis for a QS-modulated metabolic engineering study of K. pneumoniae.

摘要

群体感应 (QS) 系统普遍存在于细菌中,用于调节多种生物学功能。肺炎克雷伯氏菌是一种具有工业重要性的细菌,能够产生生物基化学品,如 2,3-丁二醇和乙酰基。该菌可以分泌呋喃基硼酸二酯 (AI-2) 作为介导 QS 系统的信号分子,该系统在次生代谢物的生物合成中发挥着关键的调节作用。在本研究中,研究了肺炎克雷伯氏菌中 QS 系统的分子调控和代谢功能。结果表明,通过敲除 luxS 破坏 AI-2 介导的 QS 后,与野生型细菌相比,肺炎克雷伯氏菌突变体中乙酰基、乙醇和乙酸的产量相对较低。然而,2,3-丁二醇的产量增加了 23.8%,达到 54.93 g/L。观察到的增强可能归因于 2,3-丁二醇脱氢酶 (BDH) 将乙酰基转化为 2,3-丁二醇的催化活性提高。这一可能性与 RT-PCR 验证的 budC 转录水平增加一致,budC 编码 BDH。这些结果还表明,肺炎克雷伯氏菌的生理代谢受到 QS 系统的不利影响。通过添加合成 AI-2 可以逆转这种影响。本研究为肺炎克雷伯氏菌的 QS 调节代谢工程研究提供了基础。

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