Partial Purification and Characterisation of Polyphenol oxidase from Two Species of Bitter Yam (Discorea dumentorum)
Biochemistry and Molecular Biology
Volume 1, Issue 2, September 2016, Pages: 11-16
Received: Jun. 1, 2016; Accepted: Jun. 12, 2016; Published: Jul. 6, 2016
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David Morakinyo Sanni, Department of Biochemistry, Federal University of Technology, Akure, Nigeria
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Polyphenol oxidases (PPO) are group of copper proteins, widely distributed phylogenetically from bacteria to mammals that catalyze the oxidation of phenolics to quinones which produces brown pigments in wounded tissues. PPO was isolated and partially characterized in white and yellow bitter yam (Discorea dumentorum). Extracts were partially purified using ammonium sulphate. The effects of optimum pH and temperature were investigated, while thermal and pH stability was also determined. The substrate specificity, kinetic and inhibition studies were also carried out. The optimal pH in the absence and presence of SDS were found to be 5.0 and 6.0, and 6.0 and 8.0 for white and yellow bitter yam respectively, while the optimum temperature for both white and yellow bitter yam was at 30°C. The enzyme was stable with over 90% activity after 3 hours incubation time at pH of 4.0 and 5.0, and 4.0 for yellow and white bitter yam respectively. The enzyme revealed over 70% remaining activities at all temperature investigated for yellow and at 30, 40 and 50°C for white bitter yam after 1 hour incubation. The Km values of 6.8 x 10-2mM and 8.0 x 10-2mM, and 3.3 x 10-1mM and 4.0 x 10-1mM with L-DOPA and tyrosine as substrates for PPO from white and yellow bitter yam respectively. Higher activity was observed with L-DOPA than with tyrosine.
Activity, Bitter Yam, Polyphenol oxidase, Partial Purification
To cite this article
David Morakinyo Sanni, Partial Purification and Characterisation of Polyphenol oxidase from Two Species of Bitter Yam (Discorea dumentorum), Biochemistry and Molecular Biology. Vol. 1, No. 2, 2016, pp. 11-16. doi: 10.11648/j.bmb.20160102.11
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Afoakwa, E. O. and Sefa-Dedeh, S. (2001). Chemical composition and quality changes in trifoliate yam Discorea dumetorum tubers after harvest. Food Chemistry, 75 (1): 88-91.
Baah, F. D., B. Maziya-Dixon, R. Asiedu, I. Oduro and W. O. Ellis (2009). Physicochemical and pasting characterisation of water yam (Discorea spp.) and relationship with eating quality of pounded yam. Journal of Food Agriculture and Environment, 7 (2): 107-112.
Bradford, M. M. (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.
Coseteng, M. Y and Lee, C. Y. (1987). Changes in apple polyphenol oxidase and polyphenol concentrations in relation to degree of browning. J. Food Sci. 52: 985-989.
D’yakov, V. L., Mishin, A. A., and Antonov, V. K. (1980). Sou. J. Biorg. Chem. 6: 157-159.
Escribano, J., Cabanes, J., Garcia-Carmona, F., (1997). Characterisation of latent polyphenol oxidase in table beet: effect of sodium dodecyl sulphate. Journal of the Science of Food and Agriculture 73: 34–38.
Flurkey, W. H. (1986). Polyphenol oxidase in higher plants. Plant Physiol. 81: 6 14-6 18.
Fraignier M., Marques L., Fleuriet A. and Macheix J., (1995). Biochemical and immunochemical characteristics of polyphenol oxidase from different fruits of Prunus. J. Agric. Food Chem. 43: 2375-2380.
Fu, Y. C., Huang, P. Y. and Chu, C. J. (2005). Use of continuous bubble separation process of separating and recovering starch and mucilage from yam (Discorea pseudojaponica Yamamoto). LWT. 38 (7): 735-744.
Fujita, S., Tono, T. and Kawahara, H. (1991). Purification and properties of polyphenol oxidase in head lettuce (Lactuca sativa). J. Sci. Food Agric. 55: 643-651.
Girardin, O., Nindjin, C., Farah, Z., Eshcher F., Stamp, P. and Otokore D. (1998). Effect of storage on system and sprout removal on post-harvest yam (Discorea spp.) fresh weight losses. Journal of Agric. Sci. 130: 329-336.
Harel, E., Mayer, A. M. and Shain, Y. (1964). Catechol oxidases from apples, their properties, subcellular location and inhibition. Physiol. Plant. 17: 921-930.
Ikediobi, C. O. and Obasuyi, H. N. (1982). Purification and some properties of O-diphenolase from white yam tubers. Phytochemistry, 21: 2815-2828.
Kenten, R. H. (1957). Latent phenolase in extracts of broad-bean (Vicia faba L.) leaves. 1. Activation by acid and alkali. Biochem. J., 67: 300-307.
Lee, C. Y., Whitaker, J. R. (Eds.) (1995). Enzymatic Browning and its Prevention. ACS Symposium Series 600. American Chemical Society, Washington, DC.
Lee, C. Y., Smith, N. L. and Pennesi, A. P. (1983). Polyphenol oxidase from DeChaunac grapes. J. Sci. Food Agric. 34: 987-991.
Marques L, Fleuriet A. and Machiex J., (1995). Characterization of multiple forms of polyphenol oxidase from apple fruit. Plant Physiol. Biochem. 33: 193-200
Mayer, A. M., (1987). Polyphenol oxidases in plants – Recents progress. Phytochemistry, 26 (1): 11–20.
Miller A. R, Kelley T J and Mujer C. V, (1990). Anodic peroxidase isoenzymes and polyphenol oxidase activity from cucumber fruit: tissue and Substrate specificity. Phytochemistry, 29: 705-709.
Moore, B. M. and Flurkey, W. H. (1990). Sodium dodecyl sulfate activation of a plant polyphenol oxidase. Effect of sodium dodecyl sulfate on enzymatic and physical characteristics of purified broad bean polyphenol oxidase. J. Biol. Chem. 265, 4982-4988.
Muzac-Tucker, I., Asemota, H. N. and Ahmad, M. H. (1993). Biochemical Composition and Storage of Jamaican Yams (Dioscorea spp). Journal of the Science of Food and Agriculture, 62: 219-224.
Nagai, T. and Suzuki, N. (2001). Partial purification of polyphenol oxidase from Chinese cabbage Brassica rapaL. J. Agric. Food Chem. 49: 3922-3926.
Ni Eidhin, D. M., Murphy, E. and O’Beirne, D. (2006). Polyphenol oxidase from apple (malus domestica borkh. cvbramley’s seedling): purification strategies and characterization. J. Food Sci. 71: 51-58.
Paul B. and Gowda L. R., (2000). Purification and Characterization of a polyphenol oxidase from the seeds of field bean (Dolichos lablab). J. Agric. Food Chem. 48, 3839-3846.
Ravindran, G. and Wanasundera, J. P. D. (1992). Chemical changes in Yam tubers (Discorea alata and Discorea esculenta) during storage. Trop. Sci. 33: 57-62.
Raymond, J., Rakariyatham, N. and Azanza, J. L. (1993). Purification and some properties of polyphenol oxidase from sunflower seeds. Phytochemistry, 34: 927-93 1.
Schrock, H. L., and Gennis, R. B. (1977). J. Biol. Chem. 252: 5990-5995
Serradell, M. D. L. A, Rozenfeld, P. A, Martinez, G. A, Civello P. M, Chaves A. R. and Anon M. C, (2000). Polyphenol oxidase activity from strawberry fruit (Fragaria xananassa, Duch.) characterization and partial purification. J. Sci. Food Agric. 80: 1421-1427
Shanti, R. A. M and Parijatham, K. M. S. (2014). Purification and characterization of polyphenol oxidase from peel and pulp of tomato. Int. J. Curr. Res. Aca. Rev. 2 (3): 74-82.
Sherman, T. D., Vaughn, K. C. and Duke, S. O. (1991). A limited survey of the phylogenetic distribution of polyphenol oxidase. Phytochemistry, 30: 2499-2506.
Swain, T., Mapson, L. W. and Robb, D. A. (1966). Activation of Vicia faba (L.) tyrosinase as affected by denaturing agents. Phytochemistry 5, 469-482.
UmitUnal M., Yabacı N., Sener A. (2011) Extraction, partial purification and characterization of polyphenol oxidase from tea leaf (Camellia sinensis). GIDA, 36 (3): 137-144.
Wong I. G, Luh B. S, Whitaker J. R (1971). Isolation and characterization of polyphénols oxidase isoenzymes of Clingstone peach. Plant Physiol. 48: 14-23.
Wesche-Ebeling, P. and Montgomery M. W., (1990). Strawberry polyphenol oxidase: Purification and characterization. J. Food Sci., 55: 1315-1319.
Wittenberg, C. and Triplett, E. L. (1985) A detergent-activated tyrosinase from Xenopuslaevis. II. Detergent activation and binding. J. Biol. Chem., 260: 12542-12546.
Yang, C., Fujita, S., Ashrafuzzaman, M., Nakamura, N. and Hayashi, N. (2000). Purification and characterization of polyphenol oxidase from banana (Musa sapientum L.) pulp. J. Agric. Food Chem., 48: 2732-2735.
Yoruk, R. and Marshall, M. R. (2003). A survey on the potential mode of inhibition for oxalic acid on polyphenol oxidase. J. Food Sci., 68: 2479-2485.
Zhou P., Smith N. L. and Lee C. Y., (1993). Potential purification and some properties of Monroe apple peel polyphenol oxidase. J. Agric. Food Chem., 41: 532-536.
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