caa a22 4500 606213317 CHVBK 20210128101047.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s10295-015-1656-z doi (NATIONALLICENCE)springer-10.1007/s10295-015-1656-z Production of C4 and C5 branched-chain alcohols by engineered Escherichia. coli [Elektronische Daten] [Xiaoyan Chen, Jingliang Xu, Liu Yang, Zhenhong Yuan, Shiyuan Xiao, Yu Zhang, Cuiyi Liang, Minchao He, Ying Guo] Higher alcohols, longer chain alcohols, contain more than 3 carbon atoms, showed close energy advantages as gasoline, and were considered as the next generation substitution for chemical fuels. Higher alcohol biosynthesis by native microorganisms mainly needs gene expression of heterologous keto acid decarboxylase and alcohol dehydrogenases. In the present study, branched-chain α-keto acid decarboxylase gene from Lactococcus lactis subsp. lactis CICC 6246 (Kivd) and alcohol dehydrogenases gene from Zymomonas mobilis CICC 41465 (AdhB) were transformed into Escherichia coli for higher alcohol production. SDS-PAGE results showed these two proteins were expressed in the recombinant strains. The resulting strain was incubated in LB medium at 37°C in Erlenmeyer flasks and much more 3-methyl-1-butanol (104mg/L) than isobutanol (24mg/L) was produced. However, in 5g/L glucose-containing medium, the production of two alcohols was similar, 156 and 161mg/L for C4 (isobutanol) and C5 (3-methyl-1-butanol) alcohol, respectively. Effects of fermentation factors including temperature, glucose content, and α-keto acid on alcohol production were also investigated. The increase of glucose content and the adding of α-keto acids facilitated the production of C4 and C5 alcohols. The enzyme activities of pure Kivd on α-ketoisovalerate and α-ketoisocaproate were 26.77 and 21.24μmolmin−1mg−1, respectively. Due to its ability on decarboxylation of α-ketoisovalerate and α-ketoisocaproate, the recombinant E. coli strain showed potential application on isoamyl alcohol and isobutanol production. Society for Industrial Microbiology and Biotechnology, 2015 3-methyl-1-butanol nationallicence Isobutanol nationallicence α-keto acid decarboxylase nationallicence Alcohol dehydrogenase nationallicence Chen Xiaoyan Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut Xu Jingliang Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut Yang Liu Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut Yuan Zhenhong Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut Xiao Shiyuan Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut Zhang Yu Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut Liang Cuiyi Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut He Minchao Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut Guo Ying Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/11(2015-11-01), 1473-1479 1367-5435 42:11<1473 2015 42 10295 https://doi.org/10.1007/s10295-015-1656-z 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/s10295-015-1656-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Chen Xiaoyan Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut NATIONALLICENCE 700 1- Xu Jingliang Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut NATIONALLICENCE 700 1- Yang Liu Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut NATIONALLICENCE 700 1- Yuan Zhenhong Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut NATIONALLICENCE 700 1- Xiao Shiyuan Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut NATIONALLICENCE 700 1- Zhang Yu Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut NATIONALLICENCE 700 1- Liang Cuiyi Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut NATIONALLICENCE 700 1- He Minchao Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut NATIONALLICENCE 700 1- Guo Ying Key Laboratory of Renewable Energy and Gas Hydrate, Chinese Academy of Sciences, 510640, Guangzhou, China aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/11(2015-11-01), 1473-1479 1367-5435 42:11<1473 2015 42 10295