Partial purification and biochemical characterization of peroxidase from rosemary (Rosmarinus officinalis L.) leaves

Authors

Department of Clinical Biochemistry, School of Pharmacy and Pharmaceutical Sciences Research Center, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: In this study, it is aimed to purify POD from leaves of Rosmarinus officinalis L. and determine its some biochemical properties. PODs are a group of oxidoreductase enzymes that catalyze the oxidation of a wide variety of phenolic compounds in the presence of hydrogen peroxide as an electron acceptor.
Materials and Methods: In this investigation, POD was purified 9.3-fold with a yield of 32.1% from the leaves of Rosemary by ammonium sulfate precipitation and ion-exchange chromatography. The enzyme biochemical properties, including the effect of pH, temperature and ionic strength were investigated with guaiacol as an electron donor. For substrate specificity investigation of the enzyme, Michaelis constant and the maximum velocity of an enzymatic reaction values for substrates guaiacol and 3,3Ͳ, 5,5Ͳ-TetraMethyle-Benzidine were calculated from the Lineweaver-Burk graphs.
Results: The POD optimum pH and temperature were 6.0 and 40°C. The POD activity was maximal at 0.3 M of sodium phosphate buffer concentration (pH 6.0). Sodium dodecyl sulphate polyacrylamide gel electrophoresis was performed for molecular weight (M w ) determination and M w of the enzyme was found to be 33 kDa. To investigate the homogeneity of the POD, native-PAGE was done and a single band was observed.
Conclusion: The stability against high temperature and extreme pH demonstrated that the enzyme could be a potential POD source for various applications in the medicine, chemical and food industries.

Keywords

1.
Nouren S, Bhatti HN, Bhatti IA, Asgher M. Kinetic and thermal characterization of peroxidase from peels of Citrus reticulate var. kinnow. J Anim Plant Sci 2013;23:430-5.  Back to cited text no. 1
    
2.
Thongsook T, Barrett DM. Purification and partial characterization of broccoli (Brassica oleracea Var. Italica) peroxidases. J Agric Food Chem 2005;53:3206-14.  Back to cited text no. 2
    
3.
O'donnell J, Huystee RV. Peanut peroxidase and the anionic component. Can J Bot 1992;70:1131-3.  Back to cited text no. 3
    
4.
Koksal E. Peroxidase from leaves of spinach (Spinacia oleracea): Partial purification and some biochemical properties. Int J Pharm 2011;7:135-9.  Back to cited text no. 4
    
5.
Vitali A, Botta B, Delle Monache G, Zappitelli S, Ricciardi P, Melino S, et al. Purification and partial characterization of a peroxidase from plant cell cultures of Cassia didymobotrya and biotransformation studies. Biochem J 1998;331 (Pt 2):513-9.  Back to cited text no. 5
    
6.
Wakamatsu K, Takahama U. Changes in peroxidase activity and in peroxidase isozymes in carrot callus. Physiol Plant 1993;88:167-71.  Back to cited text no. 6
    
7.
Biles C, Martyn R. Peroxidase, polyphenoloxidase, and shikimate dehydrogenase isozymes in relation to tissue type, maturity and pathogen induction of watermelon seedlings. Plant Physiol Biochim 1993;31:499-506.  Back to cited text no. 7
    
8.
Chanwun T, Muhamad N, Chirapongsatonkul N, Churngchow N. Hevea brasiliensis cell suspension peroxidase: Purification, characterization and application for dye decolorization. AMB Express 2013;3:14.  Back to cited text no. 8
    
9.
Cosio C, Dunand C. Specific functions of individual class III peroxidase genes. J Exp Bot 2009;60:391-408.  Back to cited text no. 9
    
10.
Schuller DJ, Ban N, Huystee RB, McPherson A, Poulos TL. The crystal structure of peanut peroxidase. Structure 1996;4:311-21.  Back to cited text no. 10
    
11.
Srinivas N, Barhate R, Raghavarao K. Aqueous two-phase extraction in combination with ultrafiltration for downstream processing of Ipomoea peroxidase. J Food Eng 2002;54:1-6.  Back to cited text no. 11
    
12.
Veitch NC. Horseradish peroxidase: A modern view of a classic enzyme. Phytochemistry 2004;65:249-59.  Back to cited text no. 12
    
13.
Angioni A, Barra A, Cereti E, Barile D, Coïsson JD, Arlorio M, et al. chemical composition, plant genetic differences, antimicrobial and antifungal activity investigation of the essential oil of Rosmarinus officinalis L. J Agric Food Chem 2004;52:3530-5.  Back to cited text no. 13
    
14.
Köksal E, Gülçin I. Purification and characterization of peroxidase from cauliflower (Brassica oleracea L. var. botrytis) buds. Protein Pept Lett 2008;15:320-6.  Back to cited text no. 14
    
15.
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680-5.  Back to cited text no. 15
    
16.
Doerge DR, Divi RL, Churchwell MI. Identification of the colored guaiacol oxidation product produced by peroxidases. Anal Biochem 1997;250:10-7.  Back to cited text no. 16
    
17.
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248-54.  Back to cited text no. 17
    
18.
Soyut H, Beydemir S. Purification and some kinetic properties of carbonic anhydrase from rainbow trout (Oncorhynchus mykiss) liver and metal inhibition. Protein Pept Lett 2008;15:528-35.  Back to cited text no. 18
    
19.
Sakharov IY, Castillo J, Areza JC, Galaev IY. Purification and stability of peroxidase of African oil palm Elaies guineensis. Bioseparation 2000;9:125-32.  Back to cited text no. 19
    
20.
Lineweaver H, Burk D. The determination of enzyme dissociation constants. J Am Chem Soc 1934;56:658-66.  Back to cited text no. 20
    
21.
Rompel A, Albers M, Naseri JI, Gerdemann C, Büldt-Karentzopoulos K, Jasper B, et al. Purification, cloning and characterization of a novel peroxidase isozyme from sweet potatoes (Ipomoea batatas). Biochim Biophys Acta 2007;1774:1422-30.  Back to cited text no. 21
    
22.
Saboora A, Parsiavash L, Moosavi-Nejad Z. Purification and kinetic properties of guaiacol peroxidase in turnip (Brassica napus var. okapi) root during different growth stages. Prog Biol Sci 2012;2:76-86.  Back to cited text no. 22
    
23.
Hu Y, Wu J, Luo P, Mo Y. Purification and partial characterization of peroxidase from lettuce stems. Afr J Biotech 2012;11:2752-8.  Back to cited text no. 23
    
24.
Lavery CB, Macinnis MC, Macdonald MJ, Williams JB, Spencer CA, Burke AA, et al. Purification of peroxidase from Horseradish (Armoracia rusticana) roots. J Agric Food Chem 2010;58:8471-6.  Back to cited text no. 24
    
25.
Bhatti HN, Najma A, Asgher M, Hanif MA, Zia MA. Purification and thermal characterization of a novel peroxidase from a local chick pea cultivar. Protein Pept Lett 2006;13:799-804.  Back to cited text no. 25
    
26.
Þiþecioðlu M, Gülçin Ý, Çankaya M, Atasever A, Þehitoðlu MH, Kaya HB, et al. Purification and characterization of peroxidase from Turkish black radish (Raphanus sativus L). J Med Plant Res 2010;4:1187-96.  Back to cited text no. 26
    
27.
Maciel HP, Gouvêa CM, Toyama M, Smolka M, Marangoni S, Pastore GM. Extraction, purification and biochemical characterization of a peroxidase from Copaifera langsdorffii leaves. Quím Nova 2007;30:1067-71.  Back to cited text no. 27
    
28.
Khatun S, Ashraduzzaman M, Karim M, Pervin F, Absar N, Rosma A. Purification and characterization of peroxidase from Moringa Oleifera L. Leaves. Bioresources 2012;7:3237-51.  Back to cited text no. 28
    
29.
Adams CD, Gorg S. Effect of pH and gas-phase ozone concentration on the decolorization of common textile dyes. J Environ Eng 2002;128:293-8.  Back to cited text no. 29
    
30.
Sakharov IY. Palm tree peroxidases. Biochemistry (Mosc) 2004;69:823-9.  Back to cited text no. 30
    
31.
Loukili AE, Limam F, Ayadi A, Boyer N, Ouelhazi L. Purification and characterization of a neutral peroxidase induced by rubbing tomato internodes. Physiol Plant 1999;105:24-31.  Back to cited text no. 31
    
32.
Pandey VP, Singh S, Singh R, Dwivedi UN. Purification and characterization of peroxidase from papaya (Carica papaya) fruit. Appl Biochem Biotechnol 2012;167:367-76.  Back to cited text no. 32
    
33.
Yemenicioglu A, Özkan M, Cemeroglu B. Partial purification and thermal characterization of peroxidase from okra (Hibiscus esculentum). J Agric Food Chem 1998;46:4158-63.  Back to cited text no. 33