Enhanced succinic acid production by Mannheimia employing optimal malate dehydrogenase.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 32327663.
- Also identified by DOI 10.1038/s41467-020-15839-z and PMC identifier 7181634.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
Succinic acid (SA), a dicarboxylic acid of industrial importance, can be efficiently produced by metabolically engineered Mannheimia succiniciproducens. Malate dehydrogenase (MDH) is one of the key enzymes for SA production, but has not been well characterized. Here we report biochemical and structural analyses of various MDHs and development of hyper-SA producing M. succiniciproducens by introducing the best MDH. Corynebacterium glutamicum MDH (CgMDH) shows the highest specific activity and least substrate inhibition, whereas M. succiniciproducens MDH (MsMDH) shows low specific activity at physiological pH and strong uncompetitive inhibition toward oxaloacetate (ki of 67.4 and 588.9 μM for MsMDH and CgMDH, respectively). Structural comparison of the two MDHs reveals a key residue influencing the specific activity and susceptibility to substrate inhibition. A high-inoculum fed-batch fermentation of the final strain expressing cgmdh produces 134.25 g L<sup>-1</sup> of SA with the maximum productivity of 21.3 g L<sup>-1</sup> h<sup>-1</sup>, demonstrating the importance of enzyme optimization in strain development.
Medical subject headings
- Bacterial Proteins
- Malate Dehydrogenase
- Pasteurellaceae
- Succinic Acid