Partial Purification, Characterization and Investigation of Inhibitory Effects of Organic Compounds on Cinnamomum verum Polyphenoloxidase Enzymes
Applied Food Biotechnology,
Vol. 7 No. 3 (2020),
10 June 2020
Background and objective: Polyphenol oxidase enzyme catalyzes oxidation of o-diphenol to o-quinone using molecular oxygen, while the final product unacceptably includes brown pigments. Therefore, inhibition of polyphenol oxidase is essential for the preservation of food products and vegetables. The enzyme is clinically beneficial for the treatment of dermal disorders with links to unusual darkening of the skin (hyper pigmentation) and is helpful in development of skincare products. The present study describes characterization and inhibition kinetics of the polyphenol oxidase from Cinnamomum verum fruit coat.
Material and methods: Purification and quantification of polyphenoloxidase were carried out using (NH4)2SO4 precipitation, dialysis with Sephadex G-100 column chromatography. The molecular weight was reported using SDS-PAGE. The Km and Vmax values were calculated using Lineweaver-Burk plot. The optimum pH, temperature and freeze-thaw were studied. Effects of several organic compounds on polyphenol oxidase activity were tested and IC50 values were calculated.
Results and conclusion: Polyphenol oxidase of Cinnamomum verum fruit coat has partially been purified as nearly 3.75-fold with an improvement of 4.58% using catechol as the substrate. The enzyme showed a single band with a molecular weight of approximately 66 kD. Optimal pH and temperature included 6.0 and 37°C, respectively. The Km and Vmax values included 1.67 Mm and 64.57 ∆A min-1, respectively. Inhibition type of cinnamic acid and ascorbic acid were uncompetitive while propyl benzoic acid showed a mixed type of inhibition. Thiol and chelating agents were strong inhibitors of enzyme activity. In conclusion, polyphenol oxidase can be used for the development of biosensors to detect and degrade phenolic compounds in industrial waste water.
- ▪ Catechol ▪ Chemical agents ▪ Cinnamomum verum ▪ Polyphenoloxidase ▪ Purification
How to Cite
- Murata M, Kurokami C, Homma S. Purification and properties of chlorogenic acid oxidase from apple (Malus pumula). Biosci. Biotech. Bioch. 1992; 56(11): 1705-1710.
- Mason H. Comparative biochemistry of the phenolase complex. Adv. Enzymol. Relat. Areas Mol. Biol. 1955; 16: 105-184.
- Mayer AM. Polyphenol oxidases in plants-Recent Progress. Phytochemistry.1986; 26(1): 11-20.
- Shiino M, Watanabe Y, Umezawa K. Synthesis of N-substituted N-nitrosohydroxylamines as inhibitors of mushroom tyrosinase. Bioorg. Med. Chem. 2001; 9(5): 1233-1240.
- Khan KM, Maharvi GM, Khan MTH, Jabbar Shaikh A, Perveen S, Begum S, Choudhary MI. Tetraketones: A new class of tyrosinase inhibitors. Bioorg. Med. Chem. 2006; 14(2): 344-351.
- Duran N, Esposito E. Potential application of oxidative enzymes and phenoloxidase-like compounds in wastewater and soil treatment: A review. Appl. Catal. B: Envir. 2000; 28: 89-99.
- Braun L. Cinnamon. J. Complement. Med.CM. 2006; 5(5): 67-68.
- Jarvill-Taylor KJ, Anderson RA, Graves DJ. A hydroxychalcone derived from cinnamon functions as a mimetic for insulin in 3T3-L1 adipocytes. J. Am. Coll. Nutr. 2001; 20(4): 327-336.
- Jayaprakasha GK, Rao LJ, Sakariah KK. Chemical composition of the volatile oil from the fruits of Cinnamomum zeylanicum blume. Flavour. Frag. J. 1997; 12(5): 331-333.
- Jayaprakasha GK, Ohnishi-Kameyama M, Ono H, Yoshida M, Jaganmohan Rao L. Phenolic constituents in the fruits of Cinnamomum zeylanicum and their antioxidant activity. J.Agr .food .Chem. 2006; 54(5): 1672-1679.
- Jayaprakasha GK, Negi PS, Jena BS, Rao LJM. Antioxidant and antimutagenic activities of Cinnamomum zeylanicum fruit extracts. J. Food Compos. Anal. 2007; 20(3-4): 330-336.
- Huang H, Zhu Q, Zhang Z, Yang B, Duan X, Jiang Y. Effect of oxalic acid on antibrowning of banana (Musa spp., AAA group, cv. ‘Brazil’) fruit during storage. Sci. Hortic. 2013;160: 208-212.
- Bevilaqua JV, Cammarota MC, Freire DMG, Sant’Anna Jr S. Phenol removal through combined biological and enzymatic treatments. Braz. J. Chem. Eng. 2002; 19(2): 151-158.
- Edalli VA, Kamanavalli CM. Removal of phenolic compounds by mushroom polyphenol oxidase from pleurotus species. The Bioscan. 2010; 4: 89 - 92.
- Flurkey A, Cooksey J, Reddy A, Spoonmore K, Rescigno A, Inlow J, Flurkey WH. Enzyme, protein, carbohydrate, and phenolic contaminants in commercial tyrosinase preparations: potential problems affecting tyrosinase activity and inhibition studies. J. Agr. Food. Chem. 2008; 56(12): 4760-4768.
- Hamed M, Shora EI, Metwally M. Use of tyrosinase enzyme from Bacillus thuringiensis for the decontamination of water polluted with phenols. Biotechnology. 2008; 7(2): 305-310.
- Hou MF, Tang XY, Zhang WD, Liao L, Wan HF. Degradation of pentachlorophenol by potato polyphenol oxidase. J. Agr. Food. Chem. 2011; 59(21): 11456-11460.
- Davis BJ. Disc electrophoresis- II method and application to human serum proteins. Ann. Ny. Acad. Sci. 1964; 121(2): 404-427.
- Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259): 680-685.
- Lineweaver H, Burk D. The determination of enzyme dissociation constants. J. Am. Chem. Soc. 1934;56(3): 658-666.
- Tohru O, Aisak GP, Kazuhito F, Koki Y. Purification and characterization of polyphenol oxidase from muscat bailey, A Grape Juice. Am. J. Enol. Vitic. 1999; 50(2): 137-143.
- Jiang Y. Role of anthocyanins, polyphenol oxidase and phenols in lychee pericarp browning. J.Sci. Food Agric. 2000; 80: 305-310.
- Bello AB, Sule MS, Alhassan AJ. Optimum pH and pH stability of crude polyphenol oxidase (PPO) extracted from five fruit samples commonly consumed in kano State, Nigeria. Bajopas. 2011; 4(1): 26-31.
- Nakamura K, Amano Y, Kagami M. Purification and some properties of a polyphenol oxidase from Koshu grapes. Am. J. Enol. Vitic.1983; 34:122-127.
- Singh A, Wadhwa N. Biochemical characterization and thermal inactivation of polyphenol oxidase from elephant foot yam (Amorphophallus paeoniifolius). J. Food Sci. Technol. 2017;54(7):2085-2093.
- Emine Z, Sule P. Purification and characterization of pear polyphenol oxidase. Turk. J. Chem. 2004; 28: 547- 557.
- Gawlik-Dziki U, Zlotek U, Swieca M. Characterization of polyphenol oxidase from butter lettuce (Lactuca sativa var. caitata L.). Food Chem. 2008; 107: 129-135.
- Anil Kumar VB, Kishor Mohan TC, Murugan K. Purification and kinetic characterization of polyphenol oxidase from Barbados Cherry (Malpighia glabra L.). Food Chem. 2008; 110: 328-333.
- Yoshiki Y, Haruyoshi K, Kazuhiko N. Polyphenol oxidase from wheat bran is a serpin. Acta Biochimica Polinica. 2008; 55: 325-328.
- Ding CK, Chachin K, Ueda Y, Mochioka R. Changes in polyphenol concentrations and polyphenol oxidase activity of loquat (Eriobotrya japonica Lindl,) fruit in relation to browning. J. Jpn. Soc. Hortic. Sci. 1998; 67: 360-366.
- Jinsen X, Tianling Z, Sadatoshi M, Shinsaku K. Purification and characterization of polyphenol oxidase from henry chestnuts (Castanea henryi). J. Wood Sci. 2004: 50: 260-265.
- Ahmet Y, Mehmet O, Bekir C. Some characteristics of polyphenol oxidase and peroxidase from Taro (Calocasia antiquorum). Tr. J. Agric. For. 1999; 23: 425-430.
- Zhongxiang F, Min Z, Yunfei S, Jingcai S. Polyphenol oxidase from bayberry (Myrica rubra Sieb. Et Zucc.): Its role in anthocyanin degradation. Food Chem. 2007; 100: 108-114.
- Ruhiye Y, Maurice RM. Physiological properties and function of plant polyphenol oxidase: A Review. J. Food Biochem. 2003; 27: 361-422.
- Lin, H, Ng AWR, Wong CW. Partial purification and characterization of polyphenol oxidase from Chinese parsley (Coriandrum sativum). Food Sci. Biotechnol.2016; 25(1): 91-96.
- Palma-Orozco G, Ortiz-Moreno A, Dorantes-Alvarez L, Sampedro JG, Najera H. Purification and partial biochemical characterization of polyphenol oxidase from mamey (Pouteria sapota). Phytochemistry. 2011; 72(1): 82-88.
- Sikora, M, Swieca M, Franczyk M, Jakubczyk A, Bochnak J, Zlotek U. Biochemical properties of polyphenol oxidases from ready-to-eat lentil (Lens culinaris Medik.) sprouts and factors affecting their activities: A search for potent tools limiting enzymatic browning. Foods. 2019; 8(5):154.1-10.
- Chikezie PC, Akuwudike AR, Chikezie CM, Ibegbulem CO. Fractional purification and kinetic parameters (Km and Vmax) of polyphenol oxidase extracted from three segments of Solanum melongenas and Musa sapietum fruits. Am. J. Plant. Physiol. 2013;7(2):1-9.
- Abstract Viewed: 98 times
- PDF Downloaded: 82 times