中国农学通报2024,Vol.40Issue(3) :95-102.

ptsG基因敲除对肺炎克雷伯氏菌CCR效应的缓解及发酵产2,3-BD的影响

ptsG Knockout:The Impact on CCR Effect of Klebsiella pneumoniae and Its Fermentation Production of 2,3-BD

毛亮阳 李娜 葛菁萍
中国农学通报2024,Vol.40Issue(3) :95-102.

ptsG基因敲除对肺炎克雷伯氏菌CCR效应的缓解及发酵产2,3-BD的影响

ptsG Knockout:The Impact on CCR Effect of Klebsiella pneumoniae and Its Fermentation Production of 2,3-BD

毛亮阳 1李娜 1葛菁萍1
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作者信息

  • 1. 黑龙江大学,生命科学学院,农业微生物技术教育部工程研究中心,黑龙江省区植物基因与生物发酵重点实验室,黑龙江省普通高校微生物重点实验室,哈尔滨 150080
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摘要

旨在解决Klebsiella pneumoniae在利用混合碳源发酵生产2,3-丁二醇(2,3-butanediol,简称2,3-BD)时会产生碳分解代谢抑制(Carbon catabolite repression,CCR)效应,导致生产效率下降的问题.本研究以Cm抗性基因为标记,采用λRed同源重组技术成功构建ptsG基因缺失菌株K.pneumoniae HD79-N.此外,在利用葡萄糖与木糖混合碳源(葡萄糖:木糖=2:1)发酵的结果显示,K.pneumoniae HD79-N菌株由于敲除ptsG基因显著减弱了 CCR效应,使其能够同步利用葡萄糖与木糖生产2,3-BD且最终产量达 9.81±0.38 g/L.同时,K pneumoniae HD79-N[0.23±0.01 g/(L·h)]菌株木糖利用率也比 K.pneumoniae HD79[0.15±0.00 g/(L·h)]菌株提高了 57.82%.本研究结果为缓解ptsG基因对K.pneumoniae菌株的CCR作用及提升2,3-BD的产量提供技术参考.

Abstract

The objective is to address the issue of Carbon Catabolite Repression(CCR)in Klebsiella pneumoniae during mixed carbon source fermentation for 2,3-butanediol(2,3-BD)production,which leads to a decrease in production efficiency.In this study,a Cm resistance gene was used as a marker,and the ptsG gene deletion strain of K.pneumoniae HD79-N was successfully constructed using the λRed homologous recombination technique.Furthermore,the fermentation results using a mixed carbon source of glucose and xylose(glucose:xylose=2:1)demonstrated that the K.pneumoniae HD79-N strain,with the ptsG gene deletion,significantly alleviated CCR,enabling simultaneous utilization of glucose and xylose for 2,3-BD production with a final yield of 9.81±0.38 g/L.Moreover,the xylose utilization rate of K.pneumoniae HD79-N strain[0.23± 0.01 g/(L·h)]was also increased by 57.82%compared to that of K.pneumoniae HD79 strain[0.15±0.00 g/(L·h)].The findings of this study provide technical insights into alleviating the CCR effect caused by the ptsG gene in K.pneumoniae strains and enhancing the production of 2,3-BD.

关键词

2,3-丁二醇/基因敲除/ptsG基因/同源重组/肺炎克雷伯氏菌

Key words

2,3-butanediol/gene knockout/ptsG gene/homologous recombination/Klebsiella pneumoniae

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基金项目

黑龙江省自然科学基金重点项目(ZD2020C008)

黑龙江省生态环境厅2022年度生态环境保护科研项目(HST2022TR004)

2023年黑龙江省高校基本科研业务费()

出版年

2024
中国农学通报
中国农学会

中国农学通报

CSTPCD
影响因子:0.891
ISSN:1000-6850
参考文献量4
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