In collaboration with Payame Noor University and Iranian Society of Physiology and Pharmacology

Document Type : Article

Authors

1 Associate Professor, Shahrekord University, Iran

2 Student of Biochemistry, Shahrekord University, Iran

Abstract

Abstract
Peroxidases are a group of oxidoreductases that are produced by a number of microorganisms and plants, and catalyse the reduction of peroxides. Peroxidases are widely used in clinical biochemistry and enzyme immunoassay. Horseradish peroxidase isoenzyme C (HRP) is one of the characterized peroxidases. The structure of the enzyme is largely alpha helical. Kinetics studies of peroxidase enzyme were performed using a spectrophotometer UV-Vis fitted with electronic control system at 35oC and 45oC and pH4 and in the presence ethanol and butanediol. Kinetic parameters show that ethanol and butanediol organic solvents becomes caused the increase of maximum speed (VMAX) and activity of the enzyme. Organic solvents have effect on electrostatic interactions of proteins because their dielectric constant with water is different. In general, reduce properties of polar solvents and decreased dielectric constant becomes caused the increases electrostatic repulsion, leading to the opening of the proteins. Enzymes activity in such organic environments increases stability, activity or facilitate reactions that are difficult to perform in aquatic environments.

Keywords

Agostini, E.; Hernandez Ruiz, J.; Arnao, M.B.; Milrad, S.R.; Tigier, H.A.; Acosta, M.; (2002). A peroxidase isoenzyme secreted by turnip (Brassica napus) hairy root cultures: inactivation by hydrogen peroxide and application in diagnostic kits. Biotechnology and applied biochemistry, 35(1): 1-7.##Biles, C.L.; Martyn, R.D.; (1993).  Peroxidase, polyphenoloxidase, and shikimate dehydrogenase isozymes in relation to tissue type, maturity and pathogen induction of watermelon seedlings. Plant physiology and biochemistry, 31(4): 499-506.##Carrea, G.; (1984). Biocatalysis in water- organic solvent two-phase systems. Trends in biotechnology, 2(4): 102-106.##Castillo, B.; Pacheco, Y.; AL-Azzam, W.; Griebenow, K.; Devi, M.; Ferrer, A.; Barletta, G.; (2005). On the activity loss of hydrolases in organic solvents: I. Rapid loss of activity of a variety of enzymes and formulations in a range of organic solvents. Journal of Molecular Catalysis B: Enzymatic, 35(4-6): 147-153.##Egorov, A.M.; Reshetnikova, I.A.; Fechina, V.A.; Gazaryan, I.G.; (1995). Comparative Studies of Plant and Fungal Peroxidases. Annals of the New York Academy of Sciences, 750(1): 469-472.##FernNdez, M.; Fragoso, A.; Cao, R.; BaOs, M.; Ansorge-Schumacher, M.; Hartmeier, W.; Villalonga, R.; (2004). Functional properties and application in peptide synthesis of trypsin  modified with  cyclodextrin – containing  dicarboxylic  acids. Journal of Molecular Catalysis B: Enzymatic, 31(1): 47-52##Fitzpatrick, P.A.; Ringe, D.; Klibanov, A.M.; (1994). X-ray crystal structure of cross-linked subtilisin Carlsberg in water vs acetonitrile. Biochemical and Biophysical Research Communications, 198(2): 675-681.##Griebenow, K.; Klibanov, A.M.; (1995). Lyophilization-induced reversible changes in the secondary structure of proteins. Proceedings of the National Academy of Sciences, 92(24): 10969-10976.##Gupta, M.N.; (1993). Enzyme function in organic solvents. EJB Reviews. Springer, 203: 25-32.##Laurenti, E.; Suriano, G.; Ghibaudi, E.M.; Ferrari, R.P.; (2000). Ionic strength and pH effect on the Fe (III)-imidazolate bond in the heme pocket of horseradish peroxidase:An EPR and UV-visible combined approach. Journal of Inorganic Biochemistry, 81(4): 259-266.##Lin, Z.; Chen, L.; Zhang, W.; (1996). Peroxidase from Ipomoea cairica (L) SW. Isolation, purification and some properties. Process Biochemistry, 31(5): 443 - 448.##Mantha, R.; Biswas, N.; Taylor, K.E.; Bewtra, J.K.; (2002). Removal of nitroaromatics from synthetic  wastewater  using two - step  zero - valent  iron  reduction and  peroxidase – catalyzed  oxidative  polymerization. Water environment research, 74(3): 280-287.##Saitou, T.; Kamada, H.; Harada, H.; (1991). Isoperoxidase in hairy roots and regenerated plants of horseradish (Armoracia lapathifolia). Plant ScienceZ 75(2): 195-201.##Simon, L.M.; Kotorman, M.; Szabo, A.; Nemcsok, J.; Laczko, I.; (2007). The effects of organic solvent/water mixtures on the structure and catalytic activity of porcine pepsin. Process Biochemistry, 42(5): 909-912.##Simon, L.M.; Laszlo, K.; Vertesi, A.; Bagi, K.; Szajani, B.; (1998). Stability of hydrolytic enzymes in water - organic solvent systems. Journal of Molecular Catalysis B: Enzymatic, 4(1-2): 41-45.##Smulevich, G.; Paoli, M.; De Sanctis, G.; Mantini, A.R.; Ascoli, F.; Coletta, M.; (1997). Spectroscopic evidence for a conformational transition in horseradish peroxidase at very low pH. Biochemistry, 36(3): 640-649.##Tayefi-Nasrabadi, H.; Keyhani, E.;  Keyhani, J.; (2006). Conformational  changes and  activity alterations induced by nickel ion in horseradish peroxidase.  Biochimie, 88(9): 1183-1197.##Veitch, N.C.; (2004). Horseradish peroxidase: a modern view of a classic enzyme. Phytochemistry, 65(3): 249-259.##Wakamatsu, K.; Takahama, U.; (1993). Changes in peroxidase activity and in peroxidase isozymes in carrot callus. Physiologia Plantarum, 88(1): 167-171.##Wang, Z.; Li, M.; Su, P.; Zhang, Y.; Shen, Y.; Han, D.; Ivaska, A.; Niu, L.; (2008). Direct electron transfer of horseradish peroxidase and its electrocatalysis based on carbon nanotube / thionine / gold composites. Electrochemistry communications, 10(2): 306-310.##