Action of Peroxidase

Britton Chance (16), using a modification of the Hartridge-Roughton flow method, studied the kinetics of the formation and dissociation of a primary peroxidase-hydrogen peroxide compound, and its reaction with acceptors like ascorbic acid and malachite green  

The reaction velocity of this compound is probably comparatively slow, since Sumner and Gjessing (66) found, for horse-radish peroxidase, that increasing the hydrogen peroxide concentration above an optimal concentration gave a successive decrease in the amoimt of purpurogallin formed. However, it was not established whether the decrease was due to the formation of compound III or to destruction of the enzyme. 

The peroxidases, in the type of reactions described above, remain in the ferric state. They are for instance not inhibited by carbon monoxide. However, the peroxidases may function as aerobic oxidases on dihydroxy-maleic acid (88). This reaction is inhibited by carbon monoxide, and the inhibition is reversed by light. This fact shows that a reversible oxidation-reduction of the peroxidase is involved in this case. Hydrogen peroxide must be formed in traces as a necessary intermediate product, since the oxygen uptake can be promptly stopped by the addition of catalase. 

The earlier literature up to 1941 has been reviewed by Sumner in the first volume of this publication. For that reason much of the older evidence need not be repeated here. 

As may be seen in Table I (page 266), several new catalases have been crystallized in the last years. The active group in catalases is proto-hematin (Zeile and Hellstrom, 94 Stem, 58). In addition to this some catalases contain a blue component which is split off by treatment with acetone and hydrochloric acid. Stem, who described the blue product first, believed it to be an impurity, whereas Sumner and Dounce (62) reported it to be a decomposition product of pure catalase. Lemberg, Norrie, and Legge (49) crystallized it and defined it as biliverdin. 

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Gierer, J. Opara, A. E. Enzymic degradation of lignin. Action of peroxidase and laccase on monomeric and dimeric model compounds. Acta Chem. Scand. 1973, 27,... 

Anni H, Yonetani T (1992) Mechanism of action of peroxidases. In Sigel H, Sigel (eds) A Metal ions in biological systems degradation of environmental pollutants by microorganisms and their metalloenzymes. Marcel Dekker, New York, p 219 Oritz de Montellano PR (1992) Annu Rev Pharmacol Toxicol 32 89 van Deurzen MP), van Rantwijk F, Sheldon RA (1997) Tetrahedron 53 13,183 Whitaker JR (1972) Principles of enzymology for the food sciences. Marcel Dekker, New York, p 591... 

LaBella F, Waykole P and Queen G (1968) Formation of insoluble gels and dityrosine by the action of peroxidase on soluble collagens. Biochem Biophys Res Comm 30, 333-338. 

It should be recognized, however, that dioxins also occur naturally, from forest fires (trees contain chloride ions) and the action of peroxidase enzymes on chlorophenols. 

Gebicki S, Gill KH, Dean RT, Gebicki JM (2002) Action of peroxidases on protein hydroperoxides. Redox Report 7 235-242... 

This was accomplished based on an understanding of the mechanism of action of peroxidase and kinetic modeling of the reaction system. 

Capsaicinoids, responsible for hot flavour, are synthesized through the cinnamic acid pathway and their degradation is aided by the action of peroxidases. The evaluation of capsaicinoids during the development, maturation and senescence of fruits of Habanero (C. chinense Jacq.), De Arbol (C. annuum var. 

Dunford HB, Stillman JS (1976) On the function and mechanism of action of peroxidases. Coord Chem Rev 19 187-251... 

Anni H, Yonetani T (1992) Mechanism of action of peroxidases. In Sigel H, Sigel A (eds) Met Ions Biol Syst, vol 28. Marcel Dekker, New York... 

In order to preserve enzyme activity during the reaction process, special attention must be paid on the substrates that cause direct or indirect enzyme inactivation. Since peroxide is a strong peroxidase inhibitor, a low peroxide/enzyme ratio must be selected. When treating phenolic compounds, the polymeric products obtained from the action of peroxidases also cause enzyme inactivation [9]. If the enzyme is inactivated, not only is the reaction hindered but, sometimes, there is a direct oxidation of the substrate by the peroxide, which causes an enantioselective reduction in some synthetic reactions [10, 11]. In these cases, an appropriate enzyme concentration and usually an adequate enzyme addition strategy are considered [8],... 

Yamazaki, Mason and Piette [63-65] have investigated the mechanism of action of peroxidases using flow ESR apparatus. The peroxidase used (from Japanese turnips) catalyses the oxidation of a number of substrates such as indoleacetic acid, dihydroxyfumarate and triose reductone by hydrogen peroxide. They were able to demonstrate directly the presence of free radical intermediates, a number of which could be identified from their hyperfine structure, and to show a correlation between ESR signal intensity and the kinetics expected for the reaction. This was strong evidence for a mechanism concerning one-electron transfer steps. The steady state concentration of free radicals was proportional to the square root of the enzyme concentration and the main decay route of the radicals was via dismutation. 

The GPD monomeric units are derived from DHCA monomer 5 via the action of peroxidase and hydrogen peroxide (Figure 5.12d). The mechanism, via a vinylogous quinone methide, involves two H-radical abstractions. Abstraction from a benzylic CH2 to produce quinone methides from phenoxy radicals has been noted previously [378]. When DHCA is subjected to peroxidase-H202, we found monomeric GPD 2, as well as the range of homo- and crossed-dimers involving DHCA and GPD. 

Phenols can be converted to polymers by the action of peroxidases and hydrogen peroxide (9.45).324... 

Chance (78) has discussed this experimental data in terms of the extended Michaelis theory which accounts for the similar peroxidatic action of peroxidase, the only difference being that with peroxidase the main reaction can proceed via the secondary complexes, whereas with catalase these complexes are inactive and the main reaction proceeds via the primary complexes. Representing the primary complexes by FeOOH and FeOOR he suggests the various reactions are ... 

Early studies of the oxidation of lAA-conjugates focused on the action of peroxidase (the classical lAA-oxidase ) on individual conjugates. Initial findings indicated that peroxidase did not attack the ester and amide conjugates tested [146] but later studies of amino acid conjugates by Park and Park [169] showed that less polar conjugates could be substrates for peroxidase. Subsequently it was found that lAA-aspartate could be oxidized by peroxidase only when peroxide was added to the reaction mixture [170] and that the product of this oxidation was 2-OH-OxIAA-aspartate. Thus the reaction with peroxidase and H2O2 yields a different product than that isolated from the plant [170]. 

Hydroxyurea (HOU) shows many biological activities and has been considered for treatment of various cancers and sickle cell disease (SOD) 39). Sickle cell patients demonstrate in vivo NO formation during HOU therapy 40). HOU mediates some of its effect via NO production. Recently, it has been explored that Ru° -pac complexes effectively catalyze the oxidation of HOU in the presence of H2O2, thus mimicking the action of peroxidase or catalase (Scheme 12) to produce NO in the reaction system 41). 

Mechanism of Action of Peroxidases Helen Anni and Takashi Yonetani... 

The most efficient of these phenolic compounds are those of the benzo-pyran class (quercitrin, ereodictyol) (Huszak, 1937). The action of peroxidase therefore resembles that of the polyphenolase in that the oxidation of ascorbic acid results from the intermediate formation of quinone derivatives. 

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