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[敲除肺炎克雷伯菌2,3-丁二醇合成关键酶基因对1,3-丙二醇产量的影响]

[Effects of knockout of 2,3-butanediol synthesis key enzyme genes on 1,3-propandediol production in Klebsiella pneumoniae].

作者信息

Guo Xinkun, Fang Huiying, Zhuge Bin, Zong Hong, Song Jian, Zhuge Jian

机构信息

Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, Jiangsu, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2013 Sep;29(9):1290-300.

Abstract

2,3-butanediol (2,3-BD) is a major byproduct of 1,3-propandediol (1,3-PDO) fermentation by Klebsiella pneumoniae. To decrease the formation of 2,3-BD, the budC and budA gene, coding two key enzymes of 2,3-BD synthetic pathway in K. pneumoniae, were knocked out using Red recombination technology. The growth of the two mutants were suppressed in different level. The budC deficient strain fermentation results showed that 1,3-PDO concentration increased to 110% and 2,3-butanediol concentration dropped to 70% of the parent strain. However, the budA deficient strain did not produce 1,3-PDO and 2,3-BD, and the final titer of lactic acid, succinic acid, ethanol and acetic acid increased remarkably compared with the parent strain. Further analysis of budC deficient strain fermentation inferred that K. pneumoniae possessed the 2,3-BD cycle as a replenishment pathway. The consequence provided a new evidence for reforming low-byproduct K. pneumoniae.

摘要

2,3-丁二醇(2,3-BD)是肺炎克雷伯菌发酵生产1,3-丙二醇(1,3-PDO)的主要副产物。为减少2,3-丁二醇的生成,利用Red重组技术敲除了肺炎克雷伯菌中编码2,3-丁二醇合成途径两种关键酶的budC和budA基因。两种突变体的生长受到不同程度的抑制。budC缺陷菌株的发酵结果表明,1,3-丙二醇浓度提高到亲本菌株的110%,2,3-丁二醇浓度降至亲本菌株的70%。然而,budA缺陷菌株不产生1,3-丙二醇和2,3-丁二醇,与亲本菌株相比,乳酸、琥珀酸、乙醇和乙酸的最终产量显著增加。对budC缺陷菌株发酵的进一步分析推断,肺炎克雷伯菌具有2,3-丁二醇循环作为补充途径。该结果为改造低副产物肺炎克雷伯菌提供了新的证据。

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