文献类型：期刊文献

英文题名：Efficient Biosynthesis of Isopropanol from Ethanol by Metabolically Engineered Escherichia coli

作者：Wang, Yuying[1];Lu, Juefeng[1];Xu, Mingcheng[1];Qian, Chen[1];Zhao, Fei[1];Luo, Yuanchan[1];Wu, Hui[1,2,3]

机构：[1]East China Univ Sci & Technol, Sch Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg Technol, Shanghai 200237, Peoples R China;[3]Key Lab Biobased Mat Engn China Natl Light Ind Cou, Shanghai 200237, Peoples R China

年份：2022

卷号：10

期号：41

起止页码：13857

外文期刊名：ACS SUSTAINABLE CHEMISTRY & ENGINEERING

收录：;EI(收录号：20224112870489);WOS:【SCI-EXPANDED(收录号:WOS:000868883100001)】；

基金：? ACKNOWLEDGMENTS This work was supported by the National Key R&D Program of China (2021YFC2103500) and the Science and Technology Commission of Shanghai Municipality (21DZ1209100) and partially supported by the Open Funding Project of the State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：ethanol utilization; isopropanol; metabolic engineering; dynamic regulation; Escherichia coli

摘要：Ethanol, a promising nonfood carbon source from syngas fermentation, has recently been used in the biosynthesis of bulk chemicals in industrial biomanufacturing. In this study, the bifunctional oxidoreductase mutant AdhEA267T/E568K was intro-duced into Escherichia coli MG1655 to construct an ethanol utilization pathway that allows its aerobic growth on ethanol. The engineered strain with ethanol utilization capacity could consume approximately 8.0 g/L of ethanol within 48 h when using 10 g/L of ethanol as the carbon source. Then, the ethanol utilization pathway was coupled with the isopropanol biosynthesis pathway to synthesize isopropanol from ethanol. The results showed that the strain carrying the ethanol-isopropanol biosynthesis pathway successfully produced 2.01 g/L of isopropanol with 10 g/L of ethanol as the sole carbon source within 48 h, and the yield was 0.20 g/g. Subsequently, this research explored two metabolic engineering strategies to enhance the productivity of isopropanol: controlling the key gene of the TCA cycle (icd) through growth-phase-dependent promoters GPPs (PrpsL/rpsJ) to dynamically regulate the growth phase and increasing NADPH supply through overexpression of pntAB. The titers of isopropanol increased to 2.44 and 4.41 g/L, respectively, with a corresponding yield of 0.24 and 0.44 g/g. The engineered E. coli strain constructed in this study can efficiently synthesize isopropanol from ethanol, providing a nonfood resource biorefinery route for industrial biomanufacturing.