Metabolic engineering of Escherichia coli W3110 to produce L-malate.
A four-carbon dicarboxylic acid L-malate has recently attracted attention due to its potential applications in the fields of medicine and agriculture. In this study, Escherichia coli W3110 was engineered and optimized for L-malate production via one-step L-malate synthesis pathway. First, deletion of the genes encoding lactate dehydrogenase (ldhA), pyruvate oxidase (poxB), pyruvate formate lyase (pflB), phosphotransacetylase (pta), and acetate kinase A (ackA) in pta-ackA pathway led to accumulate 20.9 g/L pyruvate. Then, overexpression of NADP+-dependent malic enzyme C490S mutant in this multi-deletion mutant resulted in the direct conversion of pyruvate into L-malate (3.62 g/L). Next, deletion of the genes responsible for succinate biosynthesis further enhanced L-malate production up to 7.78 g/L. Finally, L-malate production was elevated to 21.65 g/L with the L-malate yield to 0.36 g/g in a 5 L bioreactor by overexpressing the pos5 gene encoding NADH kinase in the engineered E. coli F0931 strain. This study demonstrates the potential utility of one-step pathway for efficient L-malate production. Biotechnol. Bioeng. 2017;114: 656-664. © 2016 Wiley Periodicals, Inc.
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.|Laboratory of Food Microbial-Manufacturing Engineering, Jiangnan University, Wuxi, China.
Xiaoxiang
X
Dong
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.|Laboratory of Food Microbial-Manufacturing Engineering, Jiangnan University, Wuxi, China.
Xiulai
X
Chen
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.|Laboratory of Food Microbial-Manufacturing Engineering, Jiangnan University, Wuxi, China.
Yuanyuan
Y
Qian
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.|Laboratory of Food Microbial-Manufacturing Engineering, Jiangnan University, Wuxi, China.
Yuancai
Y
Wang
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.|Laboratory of Food Microbial-Manufacturing Engineering, Jiangnan University, Wuxi, China.
Li
L
Wang
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.|Laboratory of Food Microbial-Manufacturing Engineering, Jiangnan University, Wuxi, China.
Weihua
W
Qiao
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.|Laboratory of Food Microbial-Manufacturing Engineering, Jiangnan University, Wuxi, China.
Liming
L
Liu
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019149:Bioreactors; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D008291:Malate Dehydrogenase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008293:Malates / Q000032:analysis / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods*; D009243:NAD / Q000378:metabolism; D019289:Pyruvic Acid; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D008293:Malates; D011994:Recombinant Proteins; D009243:NAD; C030298:malic acid; D019289:Pyruvic Acid; D008291:Malate Dehydrogenase
Escherichia coli; L-malate; malic enzyme; metabolic engineering