caa a22 4500 605498806 CHVBK 20210128100548.0 cr unu---uuuuu 210128e20151001xx s 000 0 eng 10.1007/s00253-015-6732-9 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6732-9 Metabolic engineering of Escherichia coli to enhance acetol production from glycerol [Elektronische Daten] [Ruilian Yao, Qing Liu, Hongbo Hu, Thomas Wood, Xuehong Zhang] Acetol, a C3 keto alcohol, is an important intermediate used to produce polyols and acrolein. To enhance acetol production from glycerol by Escherichia coli, a mutant (HJ02) was constructed by replacing the native glpK gene with the allele from E. coli Lin 43 and overexpression of yqhD, which encodes aldehyde oxidoreductase YqhD that converts methylglyoxal to acetol. Compared to the control strain without the glpK replacement, HJ02 had 5.5 times greater acetol production and a 53.4% higher glycerol consumption rate. Then, glucose was added as a co-substrate to enhance NADPH availability and the ptsG gene was deleted in HJ02 (HJ04) to alleviate carbon catabolite repression, which led to a 30% increase in the NADPH level and NADPH/NADP+. Consequently, HJ04 accumulated up to 1.20g/L of acetol, which is 69.0% higher than that of HJ02. Furthermore, the gapA gene in HJ04 was silenced by antisense RNA (HJ05) to further enhance acetol production. The acetol concentration produced by HJ05 reached 1.82g/L, which was 2.1 and 1.5 times higher than that of HJ02 and HJ04. Real-time PCR analysis indicates that glucose catabolism was rerouted from glycolysis to the oxidative pentose phosphate pathway in HJ05. Springer-Verlag Berlin Heidelberg, 2015 Acetol nationallicence Glycerol nationallicence GlpK nationallicence NADPH nationallicence GapA nationallicence Yao Ruilian Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 200240, Shanghai, China aut Liu Qing Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 200240, Shanghai, China aut Hu Hongbo Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 200240, Shanghai, China aut Wood Thomas Department of Chemical Engineering, Pennsylvania State University, 16802-4400, University Park, PA, USA aut Zhang Xuehong Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 200240, Shanghai, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/19(2015-10-01), 7945-7952 0175-7598 99:19<7945 2015 99 253 https://doi.org/10.1007/s00253-015-6732-9 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/s00253-015-6732-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Yao Ruilian Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 200240, Shanghai, China aut NATIONALLICENCE 700 1- Liu Qing Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 200240, Shanghai, China aut NATIONALLICENCE 700 1- Hu Hongbo Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 200240, Shanghai, China aut NATIONALLICENCE 700 1- Wood Thomas Department of Chemical Engineering, Pennsylvania State University, 16802-4400, University Park, PA, USA aut NATIONALLICENCE 700 1- Zhang Xuehong Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 200240, Shanghai, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/19(2015-10-01), 7945-7952 0175-7598 99:19<7945 2015 99 253