Metabolic engineering of Escherichia coli W3110 to produce L‐malate
- 17 October 2016
- journal article
- research article
- Published by Wiley in Biotechnology & Bioengineering
- Vol. 114 (3), 656-664
- https://doi.org/10.1002/bit.26190
Abstract
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. 2016;9999: 1–9.Keywords
Funding Information
- National Natural Science Foundation of China (21676118, 21422602, 31300027)
This publication has 43 references indexed in Scilit:
- Characterization of Pyruvate Uptake in Escherichia coli K-12PLOS ONE, 2013
- The Escherichia coliSLC26 homologue YchM (DauA) is a C4‐dicarboxylic acid transporterMolecular Microbiology, 2012
- One-Step Sequence- and Ligation-Independent Cloning as a Rapid and Versatile Cloning Method for Functional Genomics StudiesApplied and Environmental Microbiology, 2012
- Oxaloacetate and malate production in engineered Escherichia coli by expression of codon-optimized phosphoenolpyruvate carboxylase2 gene from Dunaliella salinaBioprocess and Biosystems Engineering, 2012
- Malic acid production from thin stillage by Aspergillus speciesBiotechnology Letters, 2011
- Engineered ketol-acid reductoisomerase and alcohol dehydrogenase enable anaerobic 2-methylpropan-1-ol production at theoretical yield in Escherichia coliMetabolic Engineering, 2011
- l -Malate Production by Metabolically Engineered Escherichia coliApplied and Environmental Microbiology, 2011
- l-Malic acid production within a microreactor with surface immobilised fumaraseMicrofluidics and Nanofluidics, 2010
- Automated design of synthetic ribosome binding sites to control protein expressionNature Biotechnology, 2009
- Malic Acid Production by Saccharomyces cerevisiae : Engineering of Pyruvate Carboxylation, Oxaloacetate Reduction, and Malate ExportApplied and Environmental Microbiology, 2008