title: Engineering Clostridium beijerinckii with the Cbei_4693 gene knockout for enhanced ferulic acid tolerance.
abstract: A mutant strain of Clostridium beijerinckii NCIMB 8052, C. beijerinckii M11, which exhibited ferulic acid tolerance up to 0.9g/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, 6.23g/L of butanol was produced in P2 medium containing 0.5g/L of ferulic acid by 4693::int, and the ferulic acid tolerance was also significantly increased up to 0.8g/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.
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech. University, No. 30, Puzhu South Road, Nanjing 211816, China; National Engineering Technique Research Center for Biotechnology, Nanjing 211816, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
authors.affiliation: Guangzhou Sugarcane Industry Research Institute, Guangdong Key Laboratory of Sugarcane Improvement and Biorefinery, Guangdong Engineering Research & Development Center for Comprehensive Utilization of Plant Fiber, Guangzhou Key Laboratory for Comprehensive Utilization of Plant Fiber, No. 10, Shiliu Gang Road, Guangzhou 510316, China. Electronic address: guoting@hotmail.com.
authors.forename: Ting
authors.initials: T
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech. University, No. 30, Puzhu South Road, Nanjing 211816, China; National Engineering Technique Research Center for Biotechnology, Nanjing 211816, China.
authors.forename: Xiaoning
authors.initials: X
authors.lastname: Shen
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech. University, No. 30, Puzhu South Road, Nanjing 211816, China; National Engineering Technique Research Center for Biotechnology, Nanjing 211816, China.
authors.forename: Jiahui
authors.initials: J
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech. University, No. 30, Puzhu South Road, Nanjing 211816, China; National Engineering Technique Research Center for Biotechnology, Nanjing 211816, China.
authors.forename: Junzhi
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech. University, No. 30, Puzhu South Road, Nanjing 211816, China; National Engineering Technique Research Center for Biotechnology, Nanjing 211816, China.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech. University, No. 30, Puzhu South Road, Nanjing 211816, China; National Engineering Technique Research Center for Biotechnology, Nanjing 211816, China.
authors.forename: Dong
authors.initials: D
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech. University, No. 30, Puzhu South Road, Nanjing 211816, China; National Engineering Technique Research Center for Biotechnology, Nanjing 211816, China.
authors.forename: Huanqing
authors.initials: H
authors.lastname: Niu
authors.affiliation: Guangzhou Sugarcane Industry Research Institute, Guangdong Key Laboratory of Sugarcane Improvement and Biorefinery, Guangdong Engineering Research & Development Center for Comprehensive Utilization of Plant Fiber, Guangzhou Key Laboratory for Comprehensive Utilization of Plant Fiber, No. 10, Shiliu Gang Road, Guangzhou 510316, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech. University, No. 30, Puzhu South Road, Nanjing 211816, China; National Engineering Technique Research Center for Biotechnology, Nanjing 211816, China. Electronic address: yinghanjie@njtech.edu.cn.
authors.forename: Hanjie
authors.initials: H
authors.lastname: Ying
mesh_terms: D046970:Clostridium beijerinckii / Q000235:genetics* / Q000378:metabolism; D003373:Coumaric Acids / Q000032:analysis / Q000378:metabolism*; D055786:Gene Knockout Techniques; D005818:Genetic Engineering / Q000379:methods*; D009249:NADP / Q000378:metabolism
chemical_list: D003373:Coumaric Acids; D009249:NADP; C004999:ferulic acid
keywords: Cbei_4693; Clostridium beijerinckii; Ferulic acid tolerance; NADPH