Hydrogen peroxide inhibition of bicupin oxalate oxidase (original) (raw)
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Oxidation reactions catalyzed by manganese peroxidase isoenzymes from Ceriporiopsis subvermispora
Febs Letters, 1995
A total of 11 manganese peroxidase isoenzymes (MnPI-MnPn) with isoelectric points (pIs) in the range of 4.58-3.20 were isolated from liquid-and solid-state cultures of the basidiomycete Ceriporiopsis subvermispora. In the presence of hydrogen peroxide, these isoenzymes showed different requirements for Mn(II) in the oxidation of vanillylacetone, o-dianisidine,p-anisidine and ABTS, whereas oxidation of guaiacol by any isoenzyme did not take place when this metal was omitted. Km values for o-dianisidine and p-anisidine in the absence of Mn(II) are in the range of 0.5-1.0 mM and 4.5-42.0 mM, respectively. Oxalate and citrate, but not tartrate, accelerate the oxidation of o-dianisidine, both in the presence and in the absence of Mn(II).
Applied Biochemistry and Biotechnology, 2002
We have previously reported the oxidation of kojic acid catalyzed by manganese peroxidase (MnP) from Ceriporiopsis subvermispora. This reaction is strictly dependent on Mn(II), although it does not require the addition of hydrogen peroxide. We have extended these studies because this reaction can be considered as a model system for the in situ generation of hydrogen peroxide in natural environments. We show here that oxidation of kojic acid with horseradish peroxidase (HRP) plus hydrogen peroxide or with manganic acetate rendered a product with identical chromatographic and spectral properties as the one obtained in the reaction catalyzed by MnP. The initial lag observed in the latter reaction decreased significantly upon UV irradiation of the substrate. On the other hand, ascorbic acid increased the lag and did not affect the yield of the reaction. The superoxide anion trapping agents glutathione, nitroblue tetrazolium, and superoxide dismutase markedly affected the reaction. In contrast, addition of the hydroxyl radical scavengers mannitol and salicylic acid had no effect. Based on these results, a mechanism for the MnP-catalyzed reaction is proposed.
Applied and Environmental Microbiology, 2005
Oxalate oxidase is thought to be involved in the production of hydrogen peroxide for lignin degradation by the dikaryotic white rot fungus Ceriporiopsis subvermispora. This enzyme was purified, and after digestion with trypsin, peptide fragments of the enzyme were sequenced using quadrupole time-of-flight mass spectrometry. Starting with degenerate primers based on the peptide sequences, two genes encoding isoforms of the enzyme were cloned, sequenced, and shown to be allelic. Both genes contained 14 introns. The sequences of the isoforms revealed that they were both bicupins that unexpectedly shared the greatest similarity to microbial bicupin oxalate decarboxylases rather than monocupin plant oxalate oxidases (also known as germins). We have shown that both fungal isoforms, one of which was heterologously expressed in Escherichia coli, are indeed oxalate oxidases that possess <0.2% oxalate decarboxylase activity and that the organism is capable of rapidly degrading exogenously supplied oxalate. They are therefore the first bicupin oxalate oxidases to have been described. Heterologous expression of active enzyme was dependent on the addition of manganese salts to the growth medium. Molecular modeling provides new and independent evidence for the identity of the catalytic site and the key amino acid involved in defining the reaction specificities of oxalate oxidases and oxalate decarboxylases.
Characterization of Ceriporiopsis subvermispora bicupin oxalate oxidase expressed in Pichia pastoris
Archives of Biochemistry and Biophysics, 2011
Oxalate oxidase (E.C. 1.2.3.4) catalyzes the oxygen-dependent oxidation of oxalate to carbon dioxide in a reaction that is coupled with the formation of hydrogen peroxide. Although there is currently no structural information available for oxalate oxidase from Ceriporiopsis subvermispora (CsOxOx), sequence data and homology modeling indicate that it is the first manganese-containing bicupin enzyme identified that catalyzes this reaction. Interestingly,
Kinetics of mediator-dependent pseudocatalatic activity of fungal peroxidases
Journal of Molecular Catalysis B: Enzymatic, 1996
The steady-state production of oxygen catalyzed by fungal Arthromyces ramosus peroxidase CARP) was investigated in the pH range from 4 to 10.6. The reaction was mediated by I-(N,N-dimethylaminek4-(4-morpholine) benzene (AMB), 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) and 1,2,4,5-tetramethoxybenzene (TMB), having low (0.39 V), medium (0.7 V) and high (0.9 V) redox potentials, respectively. AMB mediated oxygen production was indicated at pH > 7, ABTS acted at pH 2 7 and TMB was active at pH < 7. For evaluation of a mechanism of the pseudocatalatic reaction the mediators oxidation rate was measured at various pH. When AMB and ABTS were used the ARP activity decreased at pH > 8.5 and pK, of transition was 9.6 and 9.5, respectively. The TMB oxidation rate reaches a maximum at pH 5.3. The rate change at pH 4-5 corresponding to a single proton transfer with pK, 5.0 and an activity decrease at pH 5.5-7 corresponding to pK, 5.6. Analysis of the experimental results showed that oxygen was produced in a consecutive process including enzyme mediator oxidation and following chemical reaction of the oxidized mediator with hydrogen peroxide or its dissociated form (HO;). Depending on pH the oxygen production rate was limited by the chemical or the enzymatic reaction. In the range pH 4-7 oxidized TMB reacted with hydrogen peroxide (H,O,), and the process was limited by the enzymatic reaction. At pH 7 and in the more alkaline area the cation radical of ABTS reacted with H,O, and its dissociated form. Oxidized AMB reacted with HO; at pH > 7. The oxygen production rate was correlated with the reactivity of HO; estimated from the Marcus cross relationship (Marcus and Sutin, Biochim. Biophys. Acta, 811 (1985) 265).
Oxidoreductases on their way to industrial biotransformations
Biotechnology advances, 2017
Fungi produce heme-containing peroxidases and peroxygenases, flavin-containing oxidases and dehydrogenases, and different copper-containing oxidoreductases involved in the biodegradation of lignin and other recalcitrant compounds. Heme peroxidases comprise the classical ligninolytic peroxidases and the new dye-decolorizing peroxidases, while heme peroxygenases belong to a still largely unexplored superfamily of heme-thiolate proteins. Nevertheless, basidiomycete unspecific peroxygenases have the highest biotechnological interest due to their ability to catalyze a variety of regio- and stereo-selective monooxygenation reactions with H2O2 as the source of oxygen and final electron acceptor. Flavo-oxidases are involved in both lignin and cellulose decay generating H2O2 that activates peroxidases and generates hydroxyl radical. The group of copper oxidoreductases also includes other H2O2 generating enzymes - copper-radical oxidases - together with classical laccases that are the oxidore...
Description of the first fungal dye-decolorizing peroxidase oxidizing manganese(II)
Applied Microbiology and Biotechnology, 2015
Two phylogenetically divergent genes of the new family of dye-decolorizing peroxidases (DyPs) were found during comparison of the four DyP genes identified in the Pleurotus ostreatus genome with over 200 DyP genes from other basidiomycete genomes. The heterologously expressed enzymes (Pleos-DyP1 and Pleos-DyP4, following the genome nomenclature) efficiently oxidize anthraquinoid dyes (such as Reactive Blue 19), which are characteristic DyP substrates, as well as low redox-potential dyes (such as 2,2azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)) and substituted phenols. However, only Pleos-DyP4 oxidizes the high redox-potential dye Reactive Black 5, at the same time that it displays high thermal and pH stability. Unexpectedly, both enzymes also oxidize Mn 2+ to Mn 3+ , albeit with very different catalytic efficiencies. Pleos-DyP4 presents a Mn 2+ turnover (56 s −1) nearly in the same order of the two other Mn 2+-oxidizing peroxidase families identified in the P. ostreatus genome: manganese peroxidases (100 s −1 average turnover) and versatile peroxidases (145 s −1 average turnover), whose genes were also heterologously expressed. Oxidation of Mn 2+ has been reported for an Amycolatopsis DyP (24 s −1) and claimed for other bacterial DyPs, albeit with lower activities, but this is the first time that Mn 2+ oxidation is reported for a fungal DyP. Interestingly, Pleos-DyP4 (together with ligninolytic peroxidases) is detected in the secretome of P. ostreatus grown on different lignocellulosic substrates. It is suggested that generation of Mn 3+ oxidizers plays a role in the P. ostreatus white-rot lifestyle since three different families of Mn 2+-oxidizing peroxidase genes are present in its genome being expressed during lignocellulose degradation.
JBIC Journal of Biological Inorganic Chemistry, 2011
Heme peroxidases are subject to a mechanismbased oxidative inactivation. During the catalytic cycle, the heme group is activated to form highly oxidizing species, which may extract electrons from the protein itself. In this work, we analyze changes in residues prone to oxidation owing to their low redox potential during the peroxidemediated inactivation of chloroperoxidase from Caldariomyces fumago under peroxidasic catalytic conditions. Surprisingly, we found only minor changes in the amino acid content of the fully inactivated enzyme. Our results show that tyrosine residues are not oxidized, whereas all tryptophan residues are partially oxidized in the inactive protein. The data suggest that the main process leading to enzyme inactivation is heme destruction. The molecular characterization of the peroxide-mediated inactivation process could provide specific targets for the protein engineering of this versatile peroxidase.
Archives of Biochemistry and Biophysics, 1998
The kinetics of Mn 3+-oxalate formation and decay were investigated in reactions catalyzed by manganese peroxidase (MnP) from the basiomycete Ceriporiopsis subvermispora in the absence of externally added hydrogen peroxide. A characteristic lag observed in the formation of this complex was shortened by glyoxylate or catalytic amounts of Mn 3+ or hydrogen peroxide. MnP titers had a minor effect on this lag and did not influence the decay rate of the complex. In contrast, Mn 2+ and oxalate drastically affected maximal concentrations of the Mn 3+-oxalate complex formed, the decay of which was accelerated at high Mn 2+ levels. The highest concentration of complex was obtained at pH 4.0, whereas an inverse relationship was found between the pH of the reaction and the decay rate of the complex with MnP present. In the absence of MnP, the best fit for the decay kinetics of the complex gave an order of 3/2 at concentrations in the range of 30-100 µM, with a k obs = 0.012 min-1 M-0.5 at pH 4.0. The rate constant increases at lower pH values and decreases at high oxalate concentrations. The physiological relevance of these findings is discaused.