Evaluation of Inactivation Kinetics and Thermodynamic Properties of Peroxidase from Cucumis sativus for Biotechnological and Industrial Applications
Journal of Advances in Biology & Biotechnology,
The unpurified enzyme gave specific activity of 51.36 μ/mg and thereafter it was subjected to two phases of purification process of salt precipitation and gel filtration. The precipitating agent was ammonium sulphate while Sephadex-G100 served as purification matrix. The purification fold achieved after precipitation and filtration was 3.3 and with corresponding specific activities of 34.22 μ/mg and 116.31 μ/mg. The substrate used for the assay was o-dianisidine. Within 40-80°C of temperature, the kinetics of the peroxidase inactivation was evaluated. The results from assays showed that cucumber peroxidase conformed to the hypothesis of Michealis-Menten Theory. From the Lineweaver-Burk plot, Michaelis Constant (Km) and maximum velocity (Vmax) were evaluated and obtained 5.02mg/ml and 11.57μmol/min respectively. The heat induced inactivation gave biphasic curves, where initial rise in temperature was rapidly succeeded with much slower decrease. A first-order kinetic behaviour was observed for cucumber peroxidase heat inactivation. The k values of between 3.49×10-2 to 8.38×10-2 min-1 was obtained while the Z value was found to be 22.3°C. Decrease in k values with rise in temperature suggests that cucumber peroxidase was rapidly inactivated at elevated temperature. The slope of Arrhenius plot gave the activation energy of 127.99KJMol-1K-1. Also evaluated were thermodynamic constants (ΔH, G Δ, ΔS) for inactivation of peroxidase at variable temperatures. Cucumber peroxidase activity was observed to be pH sensitive and stable within pH range of 5.6-8. Further decrease or increase from this range resulted to decrease in peroxidase stability.
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Pandey VP, Manika A, Swati S, Sameeksha T, Dwivedi UN. A comprehensive review on function and application of plant peroxidases. Biochem Anal Biochem. 2017;6:308-325.
Sahin ES and Soya C. Properties of Tomato. Journal of Advancement in Plant Science. 2018; 206:1-6.
Chen LC, Chung YC, Chang CT. Characterisation of an acidic peroxidase from papaya (Carica papaya L. cv Tainung no. 2) latex and its application in the determination of micromolar hydrogen peroxide in milk. Food Chem. 2012;135:2529–2535.
Mirazizi F, Bahrami A, Haghbeen K, Zahiri HS, Bakavoli M, Legge RL. Rapid and direct spectrophotometric method for kinetics studies and routine assay of peroxidase based on aniline diazo substrates. J Enzyme Inhib Med Chem. 2016; 31:1162–1169.
Elena-Florentina G, Gabriela C, Renato F. Development of horseradish peroxidase/ layered double hydroxide hybrid catalysis for phenol degradation. Res Chem Intermediat. 2018;44(12):7731–7752.
Kondeti RS, Korivi M, Kesireddy N, Hua KC, Kesireddy SR. Protective effect of dietary ginger on antioxidant enzymes and oxidative damage in experimental diabetic rat tissues. Food Chem, 2011;124:1436–1442.
Moubasher H, Mostafa FA, Wahsh S, Haroun O. Purification and characterization of lignin peroxidase isozymes from Humicola grisea (Traaen) and its application in bioremediation of textile dyes. Egypt J Bot. 2017;57:335–343.
Mohamed SA, El-Badry MO, Drees EA, Fahmy AS. Properties of a cationic peroxidase from Citrus Jambhiri cv. Adalia. Appl Biochem Biotechnol. 2018;150:127–137
Abiodun OA. Comparative studies on nutritional composition of four melon seeds varieties. Pakistan Journal of Nutrition. 2010;9:905-908.
Khali-Ur-Rehaman M, Yaqub M.A, Sheikh S, Arshad S. Extraction and evalution of peroxidase from various vegetable sources. International Journal of Agriculture and Biology. 1999;3:170-173.
Eze SOO, Chilaka FC, Nwanguma BC. Studies on thermodynamics and kinetics of thermo-inactivation of some quality-related enzyme in white yam (Dioscorea rotunda). Journal of Thermodynamic Catalysis. 2010;1:104.
McLellan KM. and Robinson DS. Purification and heat stability of Raphanus sativus peroxidase isoenzymes. Food Chemistry, 2009;23:305-319.
Lowry HO, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry. 1951;193:265-275.
Griffin H, Dintzis FR, Krull L, L. Baker LFL. A microfibril generating factor from the enzyme complex of Trichoderma reesei. Biotechnology and Bioengeneering. 1984; 26:269–300.
Lukong B, Awah F, Nwuke C, Fobellah H. Protein isolation, purification and estimation: In: Introduction to Protein Science, Osprey Publication Center Owerri, Nigeria. 2007;55-56.
Gawlik-Dziki U, Zlotek U, Swieca M. Characterization of polyphenol oxidase from butter lettuce (Lactuca sativa). Journal of Food Chemistry. 2008;107:129-135.
Yihong H, Juan W., Ping L, Yiwei M. Purification and partial characterization of peroxidase from lettuce stems. African Journal of Biotechnology. 2012;11:2752-2756.
Melda S, lhami G, Murat C, Ali M, Habibe BK, Hasan O. Purification and characterization of peroxidase from Turkish black radish (Raphanus sativus L.). Journal of Medicinal Plants Research. 2010;4:1187-1196.
Mamounata D, Oumou HK, Nafissetou O, Romaric GB, Imael HNB, Mamoudo HD. Comparison of peroxidase activity from Allium sativum, Ipomoea batatas, Raphanus sativus and Sorghum bicolor grown in Burkina Faso. African Journal of Biochemistry Research. 2011;5:124-128.
Blee KA, Bolwell GP, Bindschedler VL, Butt V, Davies DR, Gardner SL, Gerrish C, Minibayeva F. The apoplastic oxidative burst in response to biotic stress in plant: A three component system. Journal of Experimental Botany. 2003;53:1367-1376.
Civello PM, Martinez AG, Chaves AR, Anon MC. Peroxidase from strawberry fruit: Partially purification and determination of some properties. Journal of Agriculture and Food Chemistry, 2010;43:2596-2601.
Omidiji O, Okpuzor JE, Otubu O. The contribution of an ionic peroxidase isozyme to enzyme-mediated browning in Dioscorea esculenta L. Tubers. Pakistan Journal of Nutrition. 2006;5(5): 478–480.
Ganjloo A, Rahman RA, Osman A, Baka J, Bimakr M. Kinetics of crude peroxidase inactivation and color changes of thermally treated seedless guava. Food and Bioprocess Technology. 2009;4(8):1442-1449.
Gutteridge JM. Iron promoters of the fenton reaction and lipid lipid peroxidation can be released from haemoglobin by peroxides. FEBS Lett. 1986;201:291–5.
Deepa, SS, Arumughan C. Oil palm fruit peroxidase: purification and characterization. Journal of Food Science and Technology, 2002; 39.8–13.
Maragoni AG, Jackman RL, Stanley WD. Chilling-associated softening of tomato fruit is related to increased pectinmehylesterase activity. Journal of Food Science. 1995; 60.1277–1281.
Kouadio EL, Konan HK, Dabonne S, Due DE, Kouame LP. Study of thermal stability of ɑ-Amylase sourced from digestive tract of the tropical house cricket, Gryllodes sigillatus: Kinetic and thermodynamic analysis. Journal of Novel Applied Sciences. 2013;2(3):74-82.
Kumar SR. Structure, Function and Stability of Alpha-Amylase from Jowar (Sorghum bicolar). Ph.D. Thesis, University of Myrose. 2008;99.
Chisari M, Barbagallo RN, Spagna G. Characterization of polyphenol oxidase and peroxidase and influence on browning of cold stored strawberry fruit. Journal of Agricultural and Food Chemistry. 2008;55(9):3469-3476.
Arnnok P, Ruangviriyachai C, Mahachai R, Techawongstien S, Chanthai S. Optimization and determination of polyphenol oxidase and peroxidase activities in apricots. International Food Research Journal. 2010;17(2):385-392.
Goncalves EM, Pinheiro J, Abreu M, Brandao TRS, Silva CLM. Carrot peroxidase inactivation, phenolic content and physical changes kinetics due to blanching. Journal of Food Engineering, 2013;97(4):574-581.
Siddiqui KS, Azhar MJ, Rashid MH, Rajoka MI. Stability and identification of active-site residues of carboxymethylcellulases from Aspergillus niger and Cellulomonas biazotea. Folia Microbiology. 1997;42(4): 312-318.
Saqib AN, Hassan M, Khan FN, Baig S. Thermostability of crude endoglucanase from Aspergillus fumigatus grown under solid state fermentation (SSF) and submerged fermentation (SmF). Process Biochemistry. 2010;45:641-646.
Yemenicioglu A, Ozkan M, Cemeroglu B. Thermal stabilities of peroxidases from fresh pinto beans. Journal of Food Science. 1998;63(6):987-990.
Griffin H, Dintzis, FR, Krull L, Baker FL. A microfibril generating factor from the enzyme complex of Trichoderma reesei. Biotechnology and Bioengeneering. 1984; 26:269–300.
Marin E, Sanchez L, Perez MD, Puyol P and Calvo M. Effect of heat treatment on bovine lactoperoxidase activity in skim milk: Kinetic and thermodynamic analysis. Journal of Food Sciences. 2003;68(1):89-93.
Bhatti HN, Madeeha M, Asgher M, Batool N. Purification and thermodynamic characterization of glucose oxidase from a newly isolated strain of Aspergillus niger. Canadian Journal Microbiology. 2006;52: 519-524.
Riaz M, Perveen R, Javed MR, Nadeem H, Rashi MH. Kinetics and Thermodynamic Properties of noval glucoamylase from Humicola sp. Enzyme and Microbial Technology. 2007;41:558-564.
Marin E, Sanchez L, Perez MD, Puyol P, Calvo M. Effect of heat treatment on bovine lactoperoxidase activity in skim milk: kinetic and thermodynamic analysis. Journal of Food Sciences. 2003;68(1):89-93.
Gummadi SN. What is the Role of Thermodynamics on Protein Stability? Biotechnology and Bioprocess Engineering. 2003;8:9-18.
Smith JA, Hammerschmidt R. Comparative study of acidic paroxidases associated with induced resistance in cucumber, muskmelon and watermelon. Physiological and Molecular Plant Pathology. 1998;33: 255-261.
Farrell RL, Murtagh KE, Tien M, Mozuch, MD, Kirk, TK. Physical and enzymatic properties of lignin-peroxidase isoenzymes from Phanerochaete chrysosporium. Enzyme Microbiology and Technology. 2007;11:322-328.
Whitaker JR, Lee CY. Recent advances in chemistry of enzymatic browning. Phytochemistry. 2007;29:1051-1056.
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