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

作　　者：毛亮阳 李娜 葛菁萍[1] MAO Liangyang;LI Na;GE Jingping(Engineering Research Center of Agricultural Microbiology Technology,Ministry of Education/Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region/Key Laboratory of Microbiology,College of Heilongjiang Province/School of Life Sciences,Heilongjiang University,Harbin 150080)

机构地区：[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

出　　处：《中国农学通报》2024年第3期95-102,共8页Chinese Agricultural Science Bulletin

基　　金：黑龙江省自然科学基金重点项目“乙酸代谢与副干酪乳杆菌群体感应互作探究及对细菌素产生的影响”(ZD2020C008);黑龙江省生态环境厅2022年度生态环境保护科研项目“人工重组细菌菌群对黑土磷障碍的解控机制”(HST2022TR004);2023年黑龙江省高校基本科研业务费“群体感应介导下重组根圈细菌菌群对土壤磷养分循环的调控机制”。

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。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 theλRed 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 

分 类 号：S182[农业科学—农业基础科学]