大肠杆菌乙醇工程菌mglB基因的敲除对混合糖发酵木糖利用效率的影响

doi:10.13560/j.cnki.biotech.bull.1985.2018-0962

生物技术通报 ›› 2019, Vol. 35 ›› Issue (6): 83-90.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0962

大肠杆菌乙醇工程菌mglB基因的敲除对混合糖发酵木糖利用效率的影响

许琼丹, 王永泽, 王金华, 赵筱

湖北工业大学生物工程与食品学院武汉 430068

收稿日期:2018-11-08 出版日期:2019-06-26 发布日期:2019-07-08
作者简介:许琼丹,女,硕士研究生,研究方向：发酵工程;E-mail：920705172@qq.com
基金资助:
湖北省自然科学基金项目（2015CFB637）,湖北工业大学博士科研启动基金项目（BSQD12143）

Influence of mglB Gene Knockout in Engineered Escherichia coli Producing Ethanol on the Xylose Utilization of Mix Sugar Fermentation

XU Qiong-dan, WANG Yong-ze, WANG Jin-hua, ZHAO Xiao

School of Food and Biological Engineering,Hubei University of Technology,Wuhan 430068

Received:2018-11-08 Published:2019-06-26 Online:2019-07-08

摘要/Abstract

摘要： 为明晰葡萄糖非PTS转运系统相关基因对木糖利用效率的影响,探讨葡萄糖PTS和非PTS转运系统是否对木糖利用存在协同影响,以大肠杆菌工程菌SZ470和SZ470P为出发菌株,通过RED同源重组技术敲除葡萄糖转运基因mglB,构建mglB单缺陷菌SZ470M和ptsG/mglB双缺陷菌SZ470PM。比较四株菌的混合糖（3%葡萄糖+2%木糖）发酵情况以及木糖转运代谢相关基因的转录水平。实验结果表明,SZ470M相较于出发菌株SZ470,其发酵性能无明显变化;SZ470PM的木糖消耗速度为0.37 g/L,乙醇产量为23.25 g/L,转化率为82.6%,相比于出发菌株SZ470P分别提高了32%,9.8%和5.8%。基因转录水平的分析也表明菌株SZ470P和SZ470PM的木糖转运与代谢基因的转录水平上调。综上,ptsG和mglB基因的双敲除对木糖利用效率的提高有协同影响,为促进木质纤维素作为发酵原料时木糖的高效利用提供理论依据。

关键词: 乙醇发酵, mglB基因, 混合糖发酵, 木糖利用

Abstract: To investigate the influence of glucose non-PTS transferring gene and its synergistic effect with PTS transferring gene on xylose utilization,a glucose transferring gene mglB was knocked out from engineered Escherichia coli SZ470 and its ptsG mutant SZ470P via RED homologous recombination technique,thus mglB-deleted SZ470M and ptsG/mglB-deleted SZ470P were constructed. The fermentations of mix sugar（3% glucose +2% xylose）by 4 strains were compared,and the transcriptional changes of genes related to xylose transferring and metabolism through all 4 strains were also investigated. The results showed that there were no obvious differences between SZ470M and SZ470 in fermentation. The xylose consumption rate,yield and conversion rate of ethanol by SZ470PM were 0.37 g/L,23.25 g/L and 82.6%,respectively,which increased by 32%,9.8% and 5.8% than those in starting strain SZ470P,respectively. The transcriptional analysis also demonstrated that the transcriptional levels of genes related to xylose transferring and metabolism rose in both SZ470M and SZ470PM rose. In sum,the knockout of both ptsG and mglB synergistically increase the xylose utilization rate,which contributes to the efficient utilization of xylose when using lignocelluloses materials for fermentation.

Key words: ethanol fermentation, mglB gene, mix sugar fermentation, xylose utilization

许琼丹, 王永泽, 王金华, 赵筱. 大肠杆菌乙醇工程菌mglB基因的敲除对混合糖发酵木糖利用效率的影响[J]. 生物技术通报, 2019, 35(6): 83-90.

XU Qiong-dan, WANG Yong-ze, WANG Jin-hua, ZHAO Xiao. Influence of mglB Gene Knockout in Engineered Escherichia coli Producing Ethanol on the Xylose Utilization of Mix Sugar Fermentation[J]. Biotechnology Bulletin, 2019, 35(6): 83-90.

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大肠杆菌乙醇工程菌mglB基因的敲除对混合糖发酵木糖利用效率的影响

Influence of mglB Gene Knockout in Engineered Escherichia coli Producing Ethanol on the Xylose Utilization of Mix Sugar Fermentation

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