Hydrogen peroxide inhibition of bicupin oxalate oxidase

Abstract

Oxalate oxidase is a manganese containing enzyme that catalyzes the oxidation of oxalate to carbon dioxide in a reaction that is coupled with the reduction of oxygen to hydrogen peroxide. Oxalate oxidase from Ceriporiopsis subvermispora (CsOxOx) is the first fungal and bicupin enzyme identified that catalyzes this reaction. Potential applications of oxalate oxidase for use in pancreatic cancer treatment, to prevent scaling in paper pulping, and in biofuel cells have highlighted the need to understand the extent of the hydrogen peroxide inhibition of the CsOxOx catalyzed oxidation of oxalate. We apply a membrane inlet mass spectrometry (MIMS) assay to directly measure initial rates of carbon dioxide formation and oxygen consumption in the presence and absence of hydrogen peroxide. This work demonstrates that hydrogen peroxide is both a reversible noncompetitive inhibitor of the CsOxOx catalyzed oxidation of oxalate and an irreversible inactivator. The build-up of the turnover-generated hydrogen peroxide product leads to the inactivation of the enzyme. The introduction of catalase to reaction mixtures protects the enzyme from inactivation allowing reactions to proceed to completion. Circular dichroism spectra indicate that no changes in global protein structure take place in the presence of hydrogen peroxide. Additionally, we show that the CsOxOx catalyzed reaction with the three carbon substrate mesoxalate consumes oxygen which is in contrast to previous proposals that it catalyzed a non-oxidative decarboxylation with this substrate.

Epoxidized vegetable oil and bio‐based materials as PVC plasticizer

“…Further, the increase in concentration of hydrogen peroxide and temperature affects the enzyme activity. These findings are consistent with the other researches, Goodwin et al . proved that hydrogen peroxide deactivate oxalate oxidase during the oxidation of oxalate to carbon dioxide.…”

Isolation, Screening, and Partial Characterization of Oxalate Oxidase Enzyme From Pseudomonas Strain Under Induced Oxalate Stress Condition

“…MBTH (3-methyl benzathoizaline hydrazine) used to detect release of hydrogen peroxide in the enzymatic reaction of Barley oxidase enzyme upto 1mM. In the assay of oxalate oxidase, hydrogen peroxide is produced which is coupled with HRP which catalyses the MBTH-DMA (Kanauchi et al, 2009 ;Goodwin et al, 2017).…”

“…The plate was incubated for 1hr in dark condition. The reading was noted on SYNERGY H1 microplate reader Biotek ® at 600 nm (Kanauchi et al, 2009 ;Goodwin et al, 2017).…”

“…The activity was shown in Figure9, it shows that a 60% dialyzed sample has more significant H2O2 release in the reaction compared to the standard OxO enzyme, 80% dialyzed sample, and crude enzyme(Goodwin et al, 2017;Goyal et al, 1999;Zhang et al, 1996).Some of the difficuties faced during study include the detection of H2O2 becomes unstable at higher temperature, thus the wstudy should be done at lower tempratures.…”