caa a22 4500 60550511X CHVBK 20210128100620.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00253-014-6249-7 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6249-7 Engineering Clostridium acetobutylicum with a histidine kinase knockout for enhanced n -butanol tolerance and production [Elektronische Daten] [Mengmeng Xu, Jingbo Zhao, Le Yu, I-Ching Tang, Chuang Xue, Shang-Tian Yang] Clostridium acetobutylicum JB200, a mutant strain of C. acetobutylicum ATCC 55025 obtained through strain evolution in a fibrous bed bioreactor, had high butanol tolerance and produced up to ~21g/L butanol from glucose in batch fermentation, an improvement of ~67% over the parental strain (~12.6g/L). Comparative genomic analysis revealed a single-base deletion in the cac3319 gene leading to C-terminal truncation in its encoding histidine kinase (HK) in JB200. To study the effects of cac3319 mutation on cell growth and fermentation, the cac3319 gene in ATCC 55025 was disrupted using the ClosTron group II intron-based gene inactivation system. Compared to ATCC 55025, the cac3319 HK knockout mutant, HKKO, produced 44.4% more butanol (18.2 ± 1.3 vs. 12.6 ± 0.2g/L) with a 90% higher productivity (0.38 ± 0.03 vs. 0.20 ± 0.02g/Lh) due to increased butanol tolerance, confirming, for the first time, that cac3319 plays an important role in regulating solvent production and tolerance in C. acetobutylicum. This work also provides a novel metabolic engineering strategy for generating high-butanol-tolerant and high-butanol-producing strains for industrial applications. Springer-Verlag Berlin Heidelberg, 2014 ABE fermentation nationallicence Butanol tolerance nationallicence C. acetobutylicum nationallicence Histidine kinase nationallicence Gene knockout nationallicence Metabolic engineering nationallicence Xu Mengmeng William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W. 19th Ave, 43210, Columbus, OH, USA aut Zhao Jingbo William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W. 19th Ave, 43210, Columbus, OH, USA aut Yu Le William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W. 19th Ave, 43210, Columbus, OH, USA aut Tang I-Ching Bioprocessing Innovative Company, 4734 Bridle Path Ct, 43017, Dublin, OH, USA aut Xue Chuang Department of Life Science and Biotechnology, Dalian University of Technology, 116024, Dalian, China aut Yang Shang-Tian William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W. 19th Ave, 43210, Columbus, OH, USA aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 1011-1022 0175-7598 99:2<1011 2015 99 253 https://doi.org/10.1007/s00253-014-6249-7 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6249-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Xu Mengmeng William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W. 19th Ave, 43210, Columbus, OH, USA aut NATIONALLICENCE 700 1- Zhao Jingbo William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W. 19th Ave, 43210, Columbus, OH, USA aut NATIONALLICENCE 700 1- Yu Le William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W. 19th Ave, 43210, Columbus, OH, USA aut NATIONALLICENCE 700 1- Tang I-Ching Bioprocessing Innovative Company, 4734 Bridle Path Ct, 43017, Dublin, OH, USA aut NATIONALLICENCE 700 1- Xue Chuang Department of Life Science and Biotechnology, Dalian University of Technology, 116024, Dalian, China aut NATIONALLICENCE 700 1- Yang Shang-Tian William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W. 19th Ave, 43210, Columbus, OH, USA aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 1011-1022 0175-7598 99:2<1011 2015 99 253