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Updated: Aug 9, 2026

Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins
Published on: June 20, 2019
Protein engineering of formate dehydrogenase
Vladimir I Tishkov1, Vladimir O Popov
1Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Moscow 119992, Russia. vit@enz.chem.msu.ru
Enzyme engineering improves formate dehydrogenase (FDH) for NADH regeneration in chiral compound synthesis. Modifications enhance stability and reduce costs, enabling wider industrial application of this biocatalyst.
Area of Science:
- Biocatalysis and Enzyme Engineering
- Organic Synthesis
- Biotechnology
Background:
- NAD+-dependent formate dehydrogenase (FDH) is crucial for NADH regeneration in synthesizing optically active compounds.
- Native FDHs exhibit limitations in operational stability and high production costs, hindering commercial use.
- Engineering efforts focus on overcoming these limitations for industrial applications.
Purpose of the Study:
- To review engineering strategies for bacterial and yeast FDHs.
- To enhance enzyme stability (chemical and thermal), catalytic activity, and coenzyme specificity.
- To facilitate overexpression in Escherichia coli for cost-effective production.
Main Methods:
- Enzyme engineering techniques applied to bacterial and yeast FDHs.
- Directed evolution and rational design approaches.
- Overexpression systems in Escherichia coli.
Main Results:
- Improved chemical and thermal stability of engineered FDHs.
- Enhanced catalytic activity and altered coenzyme specificity (NAD+ to NADP+).
- Successful overexpression in E. coli, potentially reducing production costs.
Conclusions:
- Enzyme engineering significantly enhances FDH performance for industrial biocatalysis.
- Engineered FDHs offer a more stable, cost-effective solution for NADH regeneration.
- These advancements pave the way for broader commercial application in chiral compound synthesis.
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