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

Metabolic engineering of Escherichia coli to produce gamma-aminobutyric acid using xylose
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大肠杆菌利用木糖生产γ-氨基丁酸的代谢工程

DOI:
10.1007/s00253-017-8162-3
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发表时间:
2017-02
影响因子:
5
通讯作者:
Wang Xiaoyuan
Wang Xiaoyuan
中科院分区:
工程技术2区
文献类型:
--
作者:
Zhao Anqi;Hu Xiaoqing;Wang Xiaoyuan

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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经济上令人感兴趣的底物。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,E.通过缺失sucA、puuE、gabT、gabP、xylA、xylB、waaC和waaF等8个基因,并过量表达2个大肠杆菌,获得了能直接生产γ-氨基丁酸的大肠杆菌菌株。大肠杆菌基因gadB和gdhA,以及五个柄杆菌新月体基因CcxylA、CcxylB、CcxylC、CcxylD和CcxylX。两个E.大肠杆菌菌株W3110和JM 109在过表达或缺失这8个基因后,都能直接从木糖中产生γ-氨基丁酸。在多重缺失突变体中的7个基因的过表达进一步增加了γ-氨基丁酸的产生。在28株重组E.大肠杆菌菌株JWZ 08/pWZt 7-g3/pWZt 7-xyl的γ-氨基丁酸产量最高。JWZ 08/pWZt 7-g3/pWZt 7-xyl以木糖为唯一碳源进行摇瓶发酵,γ-氨基丁酸产量可达3.95 g/L。
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.
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