Engineering Clostridium beijerinckii with the Cbei_4693 gene knockout for enhanced ferulic acid tolerance
Engineering Clostridium beijerinckii with the Cbei_4693 gene knockout for enhanced ferulic acid tolerance
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DOI：
10.1016/j.jbiotec.2016.04.052
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发表时间：
2016-07-10
影响因子：
4.1
通讯作者：
Ying, Hanjie
中科院分区：
文献类型：
--
作者：
Liu, Jun;Guo, Ting;Ying, Hanjie
利用大气压辉光释放和高通量筛选，获得了一株对阿魏酸耐受性高达0.9 g/L的beijerinckii M11突变株。比较基因组分析显示，该菌株携带Cbei_4693基因突变，该基因编码一种假设的蛋白质，怀疑是nadph依赖的FMN还原酶。利用ClosTron group II内含子基因失活系统对C. beijerinckii NCIMB 8052中的Cbei_4693基因进行失活后，获得了Cbei_4693基因失活突变株C. beijerinckii 4693::int。与C. beijerinckii NCIMB 8052相比，4693::int在含有0.5 g/L阿魏酸的P2培养基中产生6.23 g/L的丁醇，阿魏酸耐受性也显著提高，达到0.8 g/L。这些数据首次表明，Cbei_4693基因在贝氏弧菌发酵阿魏酸耐受性调控中起重要作用。(c) 2016 Elsevier B.V.版权所有
A mutant strain of Clostridium beijerinckii NCIMB 8052, C. beijerinckii M11, which exhibited ferulic acid tolerance up to 0.9 g/L, was generated using atmospheric pressure glow discharge and high-throughput screening. Comparative genomic analysis revealed that this strain harbored a mutation of the Cbei_4693 gene, which encodes a hypothetical protein suspected to be an NADPH-dependent FMN reductase. After disrupting the Cbei_4693 gene in C. beijerinckii NCIMB 8052 using the ClosTron group II intron-based gene inactivation system, we obtained the Cbei_4693 gene inactivated mutant strain, C. beijerinckii 4693::int. Compared with C. beijerinckii NCIMB 8052, 6.23 g/L of butanol was produced in P2 medium containing 0.5 g/L of ferulic acid by 4693::int, and the ferulic acid tolerance was also significantly increased up to 0.8 g/L. These data showed, for the first time, that the Cbei_4693 gene plays an important role in regulating ferulic acid tolerance in ABE fermentation by C. beijerinckii. (c) 2016 Elsevier B.V. All rights reserved.