Peroxidase system, model

Most of the work reported with these complexes has been concerned with kinetic measurements and suggestions of possible mechanisms. The [Ru(HjO)(EDTA)] / aq. HjOj/ascorbate/dioxane system was used for the oxidation of cyclohexanol to cw-l,3-cyclohexanediol and regarded as a model for peroxidase systems kinetic data and rate laws were derived [773], Kinetic data were recorded for the following systems [Ru(Hj0)(EDTA)]702/aq. ascorbate/dioxane/30°C (an analogue of the Udenfriend system cyclohexanol oxidation) [731] [Ru(H20)(EDTA)]70j/water (alkanes and epoxidation of cyclic alkenes - [Ru (0)(EDTA)] may be involved) [774] [Ru(HjO)(EDTA)]702/water-dioxane (epoxidation of styrenes - a metallo-oxetane intermediate was postulated) [775] [Ru(HjO)(EDTA)]7aq. H O /dioxane (ascorbic acid to dehydroascorbic acid and of cyclohexanol to cyclohexanone)... 

Recently ascorbic acid has been assigned a significant function in the reactions involving oxygen insertion by model oxygenase and peroxidase systems in which ferric ion or a ferric chelate is considered to be the catalyst. Although reaction mechanisms suggested by earlier workers involved ascorbic acid merely as a reductant to convert Fe(III) to Fe(II), which would then in turn interact with the oxidant, Hamilton... 

Scheme 5. Proposed mechanism for a model peroxidase system.

Veitch NC. 2004. Horseradish peroxidase a modem view of a classic enzyme. Phytochemistry 65 249—259. Wegrzyn TF, Farr JM, Hunter DC, Au J, Wohlers MW, Skinner MA, Stanley RA and Waterhouse DS. 2008. Stability of antioxidants in an apple polyphenol—milk model system. Food Chem 109 310-318. 

Recent work in our laboratories has confirmed the existence of a similar pathway in the oxidation of vindoline in mammals (777). The availability of compounds such as 59 as analytical standards, along with published mass spectral and NMR spectral properties of this compound, served to facilitate identification of metabolites formed in mammalian liver microsome incubations. Two compounds are produced during incubations with mouse liver microsome preparations 17-deacetylvindoline, and the dihydrovindoline ether dimer 59. Both compounds were isolated and completely characterized by spectral comparison to authentic standards. This work emphasizes the prospective value of microbial and enzymatic transformation studies in predicting pathways of metabolism in mammalian systems. This work would also suggest the involvement of cytochrome P-450 enzyme system(s) in the oxidation process. Whether the first steps involve direct introduction of molecular oxygen at position 3 of vindoline or an initial abstraction of electrons, as in Scheme 15, remains unknown. The establishment of a metabolic pathway in mammals, identical to those found in Strep-tomycetes, with copper oxidases and peroxidases again confirms the prospective value of the microbial models of mammalian metabolism concept. 

Both a synthetic lignin model substrate and the natural metabolites of the white-rot fungus were oxidized by this extracellular peroxidase. The possible roles of this nitrogen recycling system and the cinnamate pathway, which are involved in the secondary metabolism of L-phenylalanine in brown-rot and white-rot fungi, are discussed in relation to wood decay processes. 

A simplified equilibrium extraction model (Fig. 6) was presented by Dordick and colleagues [188] to explain the resolution behavior of glycoproteins in affinity based reverse micellar extraction and separation (ARMES). Their system for the study includes soybean peroxidase (SBP, MW 37 KDa, pi 4.1) and aj-acid glycoprotein (AGP, MW 43 KDa, pi 3.7) as glycoprotein solutes, concanavalin A (ConA) as the affinity ligand in AOT/isooctane RMs. The separation factor (a) for the separation of SBP from AGP can be given by... 

A typical chemical system is the oxidative decarboxylation of malonic acid catalyzed by cerium ions and bromine, the so-called Zhabotinsky reaction this reaction in a given domain leads to the evolution of sustained oscillations and chemical waves. Furthermore, these states have been observed in a number of enzyme systems. The simplest case is the reaction catalyzed by the enzyme peroxidase. The reaction kinetics display either steady states, bistability, or oscillations. A more complex system is the ubiquitous process of glycolysis catalyzed by a sequence of coordinated enzyme reactions. In a given domain the process readily exhibits continuous oscillations of chemical concentrations and fluxes, which can be recorded by spectroscopic and electrometric techniques. The source of the periodicity is the enzyme phosphofructokinase, which catalyzes the phosphorylation of fructose-6-phosphate by ATP, resulting in the formation of fructose-1,6 biphosphate and ADP. The overall activity of the octameric enzyme is described by an allosteric model with fructose-6-phosphate, ATP, and AMP as controlling ligands. 

Peroxidases are haem proteins that are activated from the ferric state to one-electron oxidants by H202. They play a significant role in the generation of radicals from xenobiotics. The compound I state contains one oxidising equivalent as an oxoferryl-haem entity and the second as a porphyrin -radical cation. Upon the oxidation of a substrate the porphyrin radical is repaired, giving the compound II. Reduction of the oxoferryl haem back to the ferric state by a second substrate molecule completes the enzyme cycle. In addition to the classical peroxidases, several other haem proteins display pseudo-peroxidase activity. The plant enzyme horseradish peroxidase (HRP) is often employed in model systems. 

Ritov, V.B., Menshikova, E.V, Goldman, R. Kagan, VE. (1996) Direct oxidation of polyunsaturated cA-parinaric fatty acid by phenoxyl radicals generated by peroxidase/H2O2 in model systems and in HL-60 cells. Toxicol. Lett., 87, 121-129... 

Commercially available horseradish peroxidase (crystalline) will substitute for luriferase in the foregoing reaction. In addition, a compound of known structure. 5-amino-2, 3-dihvdro-l, 4-phthalazinedione (also known as luminol), will substitute for luciferin. The mechanisms appear to be the same regardless of the way in which the crosses are made. Thus, a model bioluminescent system is available and can be used as a sensitivity assay for H2O2 at neutral pH. The identification of luciferase as a peroxidase is of interest since this represents the only demonstration of a bioluminescent system in which the catalytic nature of a luciferase molecule has been defined. 

---