Combining metabolic engineering and metabolic evolution to develop nonrecombinant strains of Escherichia coli C that produce succinate and malate
- 30 October 2007
- journal article
- research article
- Published by Wiley in Biotechnology & Bioengineering
- Vol. 99 (5), 1140-1153
- https://doi.org/10.1002/bit.21694
Abstract
Derivatives of Escherichia coli C were engineered to produce primarily succinate or malate in mineral salts media using simple fermentations (anaerobic stirred batch with pH control) without the addition of plasmids or foreign genes. This was done by a combination of gene deletions (genetic engineering) and metabolic evolution with over 2,000 generations of growth-based selection. After deletion of the central anaerobic fermentation genes (ldhA, adhE, ackA), the pathway for malate and succinate production remained as the primary route for the regeneration of NAD+. Under anaerobic conditions, ATP production for growth was obligately coupled to malate dehydrogenase and fumarate reductase by the requirement for NADH oxidation. Selecting strains for improved growth co-selected increased production of these dicarboxylic acids. Additional deletions were introduced as further improvements (focA, pflB, poxB, mgsA). The best succinate biocatalysts, strains KJ060(ldhA, adhE, ackA, focA, pflB) and KJ073(ldhA, adhE, ackA, focA, pflB, mgsA, poxB), produce 622–733 mM of succinate with molar yields of 1.2–1.6 per mole of metabolized glucose. The best malate biocatalyst, strain KJ071(ldhA, adhE, ackA, focA, pflB, mgsA), produced 516 mM malate with molar yields of 1.4 per mole of glucose metabolized. Biotechnol. Bioeng. 2008;99: 1140–1153.Keywords
This publication has 53 references indexed in Scilit:
- Production of l-alanine by metabolically engineered Escherichia coliApplied Microbiology and Biotechnology, 2007
- A new process for the continuous production of succinic acid from glucose at high yield, titer, and productivityBiotechnology & Bioengineering, 2007
- Low salt medium for lactate and ethanol production by recombinant Escherichia coli BBiotechnology Letters, 2006
- Methylglyoxal Bypass Identified as Source of Chiral Contamination in l(+) and d(−)-lactate Fermentations by Recombinant Escherichia coliBiotechnology Letters, 2006
- Batch culture characterization and metabolic flux analysis of succinate-producing Escherichia coli strainsMetabolic Engineering, 2006
- Betaine Tripled the Volumetric Productivity of d(-)-lactate by Escherichia coli Strain SZ132 in Mineral Salts MediumBiotechnology Letters, 2006
- Fermentation of 12% (w/v) Glucose to 1.2 m Lactate by Escherichia coli Strain SZ194 using Mineral Salts MediumBiotechnology Letters, 2006
- Development of Industrial‐Medium‐Required Elimination of the 2,3‐Butanediol Fermentation Pathway To Maintain Ethanol Yield in an Ethanologenic Strain of KlebsiellaoxytocaBiotechnology Progress, 2005
- Functional Replacement of the Escherichia coli d- (−)-Lactate Dehydrogenase Gene ( ldhA ) with the l- (+)-Lactate Dehydrogenase Gene ( ldhL) from Pediococcus acidilacticiApplied and Environmental Microbiology, 2003
- The regulation of Escherichia coli methylglyoxal synthase; a new control site in glycolysis?FEBS Letters, 1971