Assessment of Formate Dehydrogenase Stress Stability in vivo using Inactivation by Hydrogen Peroxide

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Abstract


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.

Assessment of Formate Dehydrogenase Stress Stability in vivo using Inactivation by Hydrogen Peroxide

S S Savin

A.N. Bach Institute of Biochemistry, Russian Academy of Sciences; Innovations and High Technologies MSU Ltd

V I Tishkov

A.N. Bach Institute of Biochemistry, Russian Academy of Sciences; Innovations and High Technologies MSU Ltd; M.V. Lomonosov Moscow State University

Email: vitishkov@gmail.com
Division of Chemical Enzymology, Department of Chemistry

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Copyright (c) 2010 Savin S.S., Tishkov V.I.

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