Acta NaturaeActa NaturaeActa Naturae2075-8251Acta Naturae Ltd1077510.32607/20758251-2010-2-1-97-101ArticlesСтатьиResearch ArticleAssessment of Formate Dehydrogenase Stress Stability in vivo using Inactivation by Hydrogen PeroxideAssessment of Formate Dehydrogenase Stress Stability in vivo using Inactivation by Hydrogen PeroxideSavinS STishkovV I

Division of Chemical Enzymology, Department of Chemistry

vitishkov@gmail.comA.N. Bach Institute of Biochemistry, Russian Academy of SciencesInnovations and High Technologies MSU LtdA.N. Bach Institute of Biochemistry, Russian Academy of SciencesInnovations and High Technologies MSU LtdM.V. Lomonosov Moscow State University1503201021VOL 2, NO1 (2010)ТОМ 2, №1 (2010)9710117012020Copyright ©; 2010, Savin S.S., Tishkov V.I.Copyright ©; 2010, Savin S.S., Tishkov V.I.2010Savin S.S., Tishkov V.I.Savin S.S., Tishkov V.I.https://creativecommons.org/licenses/by/4.0https://actanaturae.ru/2075-8251/article/view/10775

Kinetic studies on hydrogen peroxide-induced inactivation of mutant formate dehydrogenase from Pseudomonas sp. 101 (PseFDH Cys255Ala) suggest a simple bimolecular mechanism for enzyme reaction with the inactivation agent. In the excess of hydrogen peroxide, the decrease in enzyme activity follows first-order kinetics. Therefore, the first-order effective inactivation kinetic constants determined for various FDH forms at a constant H 2O 2 concentration can be used as a quantitative measure of the enzyme stability. It was shown that two cysteine residues located in the active site formate- and coenzyme-binding domains (Cysl45 and Cys255, respectively) make similar contributions to the enzyme stability, while the contribution of Cys354 is insignificant. The inactivation kinetics of wild-type PseFDH, mutant PseFDH Cysl45Ser/Cys255Ala, and FDH produced under stress conditions by bacterium Staphylococcus aureus, higher plants Arabidopsis thaliana, and soya Glycine max, was studied. It was found that the stress-induced FDHs are at least 20 times more stable than the nonstress-induced PseFDH from Pseudomonas sp. 101 grown on methanol.

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