Abstract
In the previous work, after screening tropical plants (43 species) for peroxidase activity, high activity has been detected in leaves of some palms and especially African oil palm Elaeis guineensis. This palm is widely cultivated in Colombia and presents a promising source for the industrial production of peroxidase. The initial enzyme isolation included homogenization and extraction of pigments using aqueous two phase polymer system. Initially, traditional system, formed by polyethyleneglycol/K2HPO4, was used. The replacement of K2HPO4 with (NH4)2SO4 allowed direct application of the salt phase with accumulated peroxidase on a Phenyl-Sepharose column. The final purification was carried out by liquid chromatography on Sephacryl S200 and DEAE-Toyopearl columns. The specific activity of the purified peroxidase measured toward guaiacol was 4300 units per mg of protein. The molecular weight and isoelectric point for palm peroxidase were 57.000 and 3.8, respectively. Palm peroxidase possesses uniquely high thermostability and is more stable in organic solvents than horseradish peroxidase is.
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References
Adam W, Lazarus M, Saha-Moller CR, Weichold O, Hoch U and Schreier P (1999), Biotransformations with peroxidases. Adv. Biochem. Eng. 63: 74-108
Albertson P-Å (1986) Partition of Cell Particles and Macromolecules. Wiley, New York.
Berezin IV, Kerschengoltz BM and Ugarova NN (1975) Microenvironment of enzymes as one of factors defining an enzymatic stability. Stability of soluble and immobilized horseradish peroxidase. Dokl. Acad. Sci. USSR 223: 1256-1259
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254
Clemente E (1998) Purification and thermostability of isoperoxidase from oranges. Phytochemistry 49: 29-36
Dordick S, Marletta MA and Klibanov AM (1987) Polymerization of phenoles catalyzed by peroxidase in nonaqueous media. Biotech. Bioeng. 30: 31-36
Dunford HB (1991) Horseradish peroxidase: structure and kinetic properties. In: Everse J, Everse KE, Grisham MB (eds.) Peroxidase in Chemistry and Biology (pp. 1-24) CRC Press, Boca Raton, FL.
Farrell RL, Murtagh KE, Tien M, Mozuch MD and Kirk TK (1989) Physical and enzymatic properties of lignin-peroxidase isoenzymes from Phanerochaete chrysosporium. Enzyme Microb. Technol. 11: 322-328
Gazaryan IG and Lagrimini LM (1996) Purification and unusual kinetic properties of a tobacco anionic peroxidase. Phytochemistry 41: 1029-1034
Gazaryan IG, Reshetnikova IA, Doseeva VV and Bekker EG (1995) Isolation and comparative analysis of peroxidases from the fungus Phellinus igniarius (L.:Fr) quel. 90-1 and tobacco. Biochemistry (Moscow) 60: 767-771
Gorton L (1995) Carbon paste electrodes modified with enzymes, tissues, and cells. Electroanalysis 7: 23-45
Krell H-W (1991) Peroxidase. An important enzyme for diagnostic test kits. In: Lobarsewski J, Greppin H, Penel C and Gaspar Th (eds.) Biochemical, Molecular and Physiological Aspects of Plant Peroxidases (pp. 469-478) Univ.M. Curie-Sklodowska and Univ. Geneva, Lublin and Geneva.
Laemmli UK (1970) Cleavage of structural proteins durind the assembly of the head of bacteriophage T4. Nature 227: 680-685
Mader M (1992) Plant Peroxidases 1980–1990. In: Penel C, Gaspar Th and Greppin H (eds.) Topics and Detailed Literature on Molecular, Biochemical and Physiological Aspects (pp. 37-46) Univ. Geneva, Geneva.
Marangoni AG, Brown ED, Stanley DW and Yada RY (1989) Tomato peroxidase: rapid isolation and partial characterization. J. Food Sci. 54: 1269-1271
Miland E, Smyth MR and Fagin CO (1996) Phenol removal by modified peroxidases. J. Chem. Tech. Biotechnol. 67: 227-236.
Obinger C, Burner U, Ebermann R, Penel C and Greppin H (1996) Plant Peroxidases. Biochemistry and Physiology. Geneva-Vienna. Univ. Agriculture and Univ. Geneva, Vienna and Geneva.
Saitoh T, Tani H, Kamidate T and Watanabe H (1995) Phase separation in aqueous micellar solutions of nonionic surfactants for protein separation. Trends Anal. Chem. 14: 213-217.
Sakharov I Yu, Litvin FE and Artyukov AA (1994) Purification and characterization of two serine collagenolytic proteases from crab Paralithodes camschatica. Comp. Biochem. Physiol. 108B: 561-568.
Sakharov I Yu and Bautista AG (1999) Variation of peroxidase activity in cocoa (Theobroma cacao L.) beans during their ripening, fermentation and drying. Food Chem. 65: 51-54
Sakharov I Yu, Bautistaw G, Sakharova IV, Rojas A and Pletyuschkina, O Yu (1999) Peroxidase in tropical plants. Revista Colombiana de Quimica 28: 97-106
Scopes RK (1994), Protein purification: principles and practice, Springer-Verlag, New York.
Tijssen P (1985) Practice and Theory of Enzyme Immunoassay, Elsevier, Amsterdam.
Vitali A, Botta B, Monache G, Zappitelli S, Ricciardi P, Melino S, Petruzzelli R and Giardina B (1998) Purification and partial characterization of a peroxidase from plant cell cultures of Cassia didymobotrya and biotransformation studies. Biochem. J. 331: 513-519
Wakamatsu K and Takahama U (1993) Changes in peroxidase activity and peroxidase isoenzymes in carrot callus. Physiol. Plants 88: 167-171
Welinder KG, Rasmussen SK, Penel C and Greppin H (1993) Plant Peroxidases. Biochemistry and Physiology, Univ. of Copenhagen and Univ. Geneva, Copenhagen and Geneva.
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Sakharov, I., Castillo, J.L., Areza, J. et al. Purification and stability of peroxidase of African oil palm Elaies guineensis. Bioseparation 9, 125–132 (2000). https://doi.org/10.1023/A:1008117217885
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DOI: https://doi.org/10.1023/A:1008117217885