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利用木糖对大肠杆菌进行代谢工程改造以生产γ-氨基丁酸。

Metabolic engineering of Escherichia coli to produce gamma-aminobutyric acid using xylose.

作者信息

Zhao Anqi, Hu Xiaoqing, Wang Xiaoyuan

机构信息

State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 xLihu Avenue, Wuxi, 214122, China.

School of Biotechnology, Jiangnan University, Wuxi, 214122, China.

出版信息

Appl Microbiol Biotechnol. 2017 May;101(9):3587-3603. doi: 10.1007/s00253-017-8162-3. Epub 2017 Feb 11.

Abstract

Biomass-derived xylose is an economically interesting substrate for the sustainable microbial production of value-added compounds. Escherichia coli could barely use xylose to directly produce gamma-aminobutyric acid. In this study, E. coli strains that could directly produce gamma-aminobutyric acid were developed through the deletion of eight genes sucA, puuE, gabT, gabP, xylA, xylB, waaC, and waaF, and the overexpression of two E. coli genes gadB and gdhA, as well as five Caulobacter crescent genes CcxylA, CcxylB, CcxylC, CcxylD, and CcxylX. Both E. coli strains W3110 and JM109 could directly produce gamma-aminobutyric acid from xylose after either overexpression of the seven genes or deletion of the eight genes. Overexpression of the seven genes of in the multiple deletion mutants further increased gamma-aminobutyric acid production. Among the 28 recombinant E. coli strains constructed in this study, the highest gamma-aminobutyric acid was produced by JWZ08/pWZt7-g3/pWZt7-xyl. JWZ08/pWZt7-g3/pWZt7-xyl could produce 3.95 g/L gamma-aminobutyric acid in flask cultivation, using xylose as the sole carbon source.

摘要

生物质衍生的木糖是用于可持续微生物生产增值化合物的具有经济吸引力的底物。大肠杆菌几乎不能利用木糖直接生产γ-氨基丁酸。在本研究中,通过缺失8个基因sucA、puuE、gabT、gabP、xylA、xylB、waaC和waaF,以及过表达两个大肠杆菌基因gadB和gdhA,还有5个新月柄杆菌基因CcxylA、CcxylB、CcxylC、CcxylD和CcxylX,构建了能够直接生产γ-氨基丁酸的大肠杆菌菌株。在过表达这7个基因或缺失那8个基因后,大肠杆菌菌株W3110和JM109都能够从木糖直接生产γ-氨基丁酸。在多重缺失突变体中过表达这7个基因进一步提高了γ-氨基丁酸的产量。在本研究构建的28株重组大肠杆菌菌株中,JWZ08/pWZt7-g3/pWZt7-xyl产生的γ-氨基丁酸最多。JWZ08/pWZt7-g3/pWZt7-xyl在以木糖为唯一碳源的摇瓶培养中能够产生3.95 g/L的γ-氨基丁酸。

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