4.0 Article

Effect of Additional Amino Acid Replacements on the Properties of Multi- point Mutant Bacterial Formate Dehyderogenase PseFDH SM4S

Journal

ACTA NATURAE
Volume 14, Issue 1, Pages 82-91

Publisher

RUSSIAN FEDERATION AGENCY SCIENCE & INNOVATION
DOI: 10.32607/actanaturae.11665

Keywords

formate dehydrogenase; Pseudomonas sp. 101; catalytic properties; thermal stability; site-directed mutagenesis

Funding

  1. Russian Federation [MD-349.2021.1.4]

Ask authors/readers for more resources

A mutant formate dehydrogenase (PseFDH) from Pseudomonas sp. 101 bacterium with improved thermal stability has been generated through single-point amino acid replacements. The mutant with E170D substitution exhibits increased thermal stability compared to the starting mutant and the wild-type enzyme.
Formate dehydrogenase from Pseudomonas sp. 101 bacterium (PseFDH, EC 1.2.1.2) is a research model for the elucidation of the catalytic mechanism of 2-oxyacid D-specific dehydrogenases enzyme superfamily. The enzyme is actively used for regeneration of the reduced form of NAD(P)H in chiral synthesis with oxidoreductases. A multi-point mutant PseFDH SM4S with an improved thermal and chemical stability has been prepared earlier in this laboratory. To further improve the properties of the mutant, additional single-point replacements have been introduced to generate five new PseFDH mutants. All new enzymes have been highly purified, and their kinetic properties and thermal stability studied using analysis of thermal inactivation kinetics and differential scanning calorimetry. The E170D amino acid change in PseFDH SM4S shows an increase in thermal stability 1.76- and 10-fold compared to the starting mutant and the wild-type enzyme, respectively.

Authors

I am an author on this paper
Click your name to claim this paper and add it to your profile.

Reviews

Primary Rating

4.0
Not enough ratings

Secondary Ratings

Novelty
-
Significance
-
Scientific rigor
-
Rate this paper

Recommended

No Data Available
No Data Available