MSRT,ISC
37
h-index
h-index
Partial Purification and Characterisation of Polyphenol Oxidase from Tomatoes (Solanum Lycopersicum)
Document Type : Original Article
Author
Assistan professor of biochemistry, Department of Biology, Payame noor university, Iran
Abstract
Polyphenol oxidase (PPO) from tomatoes was extracted and partially purified through (NH4)2SO4 precipitation, dialysis and ion exchange chromatography. The activity of polyphenol oxidase was investigated in solanum lycopersicum. Spectrophotometric method was used to assay the enzyme activity and the kinetic constants - maximum enzyme velocity (Vmax) and Michealis - Menten constant (Km). Of the substrates tested, pyrogallol was the best substrate for PPO with a Km value of 1.5 mM. The optimum pH for PPO activity was found to be 6.8. The enzyme showed high activity over a broad pH range of 4 - 8. The optimal pH and temperature for enzyme activity were found to be 6.8 and 50-60 °C, respectively. km value for tomatoe PPO is calculated 25 mM for catechol and 1.5 mM for pyrogallol and 8.5 mM for L-dopa. As can be seen, affinity of PPOs for various substrates varies widely. The enzyme showed a broad activity over a broad pH and temperature range. The thermal inactivation studies showed that the enzyme is heat resistant. The enzyme showed the highest activity toward pyrogallol and no activity toward tyrosine. Of the inhibitors tested, the most potent inhibitors was sodium kojic acid
Keywords
Broothaerts W, Mcpherson J.LI.B, Randall E, Lane, W.D, Wierma P.A. (2000), Fast apple (Malus x domestica) and tobacco (Nicotiana tobacum) leaf polyphenol oxidase activity assay for screening transgenic plants. J. Agric. Food Chem. 48, pp. 5924-5928.
Ding CK, Chachin K, Ueda Y, Imahori Y. (1998). Purification and properties of
polyphenol oxidase from loquat fruit. J Agric Food Chem, 46:4144-4149.
Dogan S, Turan Y, Ertürk H and Arslan O. (2005), Characterization and purification of polyphenol oxidase from Artichoke (Cynara scolymus L.). Journal of Agriculture and Food Chemistry, 53, 776-785.
Dogan M, Arslan O, Dogan S. (2002). Substrate specificity, heat inactivation and inhibition of polyphenol oxidase from different aubergine cultivars. Int J Food Sci Technol, 37:415-423.
Duangmal K and Apenten RKO (1999). A comparative study of polyphenoloxidases from taro (Colocasia esculenta) and potato (Solanum
tuberosum var. Romano). Food Chem, 64:351-359.
Golbeck J.H, Cammarata K.V. (1981). Spinach Thylakoid Polyphenol Oxidase Isolation, activation and properties of the native chloroplast enzyme. Plant Physiol. 67, pp. 977-984.
Gandia-Herrero F, Garcia-Carmona F, Escribano J. (2004). Purification and characterization of a latent polyphenol oxidase from beet root (Beta vulgaris L.). J. Agric. Food Chem. 52(3), pp. 609-615.
Gao J, Wang B, Feng X, Tang H, LI W and Zhang K. (2011). Partial Properties of Polyphenol Oxidase in Sour Cherry (Prunus cerasus L. CV. CAB) Pulp, World Journal of Agricultural Sciences 7 (4): 444-449, 2011.
Gawlik-Dziki U, Szymanowska U, Baraniak B. ( 2007). Characterization of
polyphenol oxidase from broccoli (Brassica oleracea var. botrytis italica)
florets. Food Chem, 105:1047-1053.
Gomez-Lopez VM. (2002). Some biochemical properties of polyphenol oxidase from two varieties of avocado. Food Chem, 77:163-169.
Gonzalez E M, Ancos B and Cano M.P. (2000), Partial characterization of peroxidase and polyphenol oxidase activities in Blackberry fruits. Journal of Agriculture and Food Chemistry, 48, 5459-5464.
Halder J, Tamuli P and Phaduri A.N. (1998), Isolation and characterization of polyphenol oxidase from Indian tea leaf (Camellia sinensis). Journal of Nutritional Biochemistry, 9, 75-80.
Jimenezatienzar M, Pedreno M, Garciacarmona F. (1991). Activation of polyphenol oxidase by Polyamines. Biochem. Int. 25, pp. 861.
Kavrayan D and Aydemir T. (2001), Partial purification and characterization of
polyphenoloxidase from peppermint (Mentha piperita). Food Chemistry, 74, 147-154.
Laveda F, Nunez-Delicado E, Garcia-Carmona F, Sanchez-Ferrer A. (2001). Proteolytic activation of latent Paraguaya peach PPO. Characterization of monophenolase activity. J. Agric. Food Chem. 49(2), pp. 1003-1008.
Lourenço E.J, Leão J.S and Neves V.A. (1990), Heat inactivation and kinetics of polyphenoloxidase from palmito (Euterpe edulis). Journal of Science Food and Agriculture, 52, 249-259.
Lourenço E.J, Neves V.A and Da Silva M.A. (1992). Polyphenoloxidase from sweet potato: Purification and properties. Journal of Agricultural and Food Chemistry, 40, 2369-2373.
Lowry O.H, A. L. Farr, Rosebrough, and R. J. Randall, “Protein measurement with the FolinPhenol Matheis, G., Whitaker, J.R. 1984, Modification of proteins by polyphenol oxidase and peroxidase and their products. J. Food Biochem. 8(3), pp. 137-162.
Okot-Kotber M, Liavoga, A, Yong K.J, Bagorogoza K. (2002). Activation of polyphenol oxidase in extracts of bran from several wheat (Triticum aestivum) cultivars using organic solvents, detergents, and chaotropes. J. Agric. Food Chem. 50(8), pp. 2410-2417.
Rapeanu G, Van Loey A, Smout C and Hendrickx M. (2006). Biochemical characterization and process stability of polyphenoloxidase extracted from Victoria grape (Vitis vinifera ssp. Sativa). Food Chemistry, 94, 253-261.
ROBB D.A. (1984). Copper Protein and Copper enzyme. In: Contie R (ed). Vol 2. pp 207-241.
CRC Press. Boca Raton, FL. Robinson DS, Eskin NAM (eds), Zawistowski J, Biliaderis CG, Eskin NAM. (1991). Oxidative Enzymes in Foods. Elsevier, New York, pp. 217-273.
Sanchez-Amat A, Solano F. (1997). A pluripotent polyphenol oxidase from the melanogenic marine alteromonas shares catalytic capabilities. Biochem .Biophys. Res. Comm. 240, pp. 787-792.
Santosh R.K, Beena P, Appu Rao A.G, GOWDA. L.R. (2006). The conformational state of polyphenol oxidase from field bean (Dolichos lablab) upon SDS and acid-pH activation. Biochem. J. 395(3), pp. 551-562.
Serradell MA, Rozenfeld PA, Martinez GA, Civello PM, Chaves AR, Anon MC. (2000). Polyphenoloxidase activity from strawberry fruit (Fragaria x ananassa, Duch., cv Selva): characterization and partial purification. J Sci Food Agric, 80:1421-1427.
Ünal MÜ, fiener A. (2006). Determination of biochemical properties of polyphenol oxidase from Emir grape (Vitis vinifera L. cv. Emir). J Sci Food Agric, 86:2374-2379.
Ünal MÜ. (2007). Properties of polyphenol oxidase from Anamur banana (Musa cavendishii).Food Chem, 100:909-913.
Van Gelder C.W, Flurkey W.H, Wichers H.J. (1997). Sequence and structural features of plant and fungal tyrosinases. Phytochemistry. 45, pp.1309-1323.
Vamos-Vigyazo, L. (1981). Polyphenol oxidase and peroxidase in fruits and
vegetables. Crit Rev Food Sci, 15:49-127.
Yang C-P, Fujita S, Kohno K, Kusubayashi A, Ashrafuzzaman MD, Hayashi N.
( 2001) .Partial purification and characterization of polyphenol oxidase from banana (Musa sapientum L) peel. J Sci Food Agric, 49:1446-1449.
Ding CK, Chachin K, Ueda Y, Imahori Y. (1998). Purification and properties of
polyphenol oxidase from loquat fruit. J Agric Food Chem, 46:4144-4149.
Dogan S, Turan Y, Ertürk H and Arslan O. (2005), Characterization and purification of polyphenol oxidase from Artichoke (Cynara scolymus L.). Journal of Agriculture and Food Chemistry, 53, 776-785.
Dogan M, Arslan O, Dogan S. (2002). Substrate specificity, heat inactivation and inhibition of polyphenol oxidase from different aubergine cultivars. Int J Food Sci Technol, 37:415-423.
Duangmal K and Apenten RKO (1999). A comparative study of polyphenoloxidases from taro (Colocasia esculenta) and potato (Solanum
tuberosum var. Romano). Food Chem, 64:351-359.
Golbeck J.H, Cammarata K.V. (1981). Spinach Thylakoid Polyphenol Oxidase Isolation, activation and properties of the native chloroplast enzyme. Plant Physiol. 67, pp. 977-984.
Gandia-Herrero F, Garcia-Carmona F, Escribano J. (2004). Purification and characterization of a latent polyphenol oxidase from beet root (Beta vulgaris L.). J. Agric. Food Chem. 52(3), pp. 609-615.
Gao J, Wang B, Feng X, Tang H, LI W and Zhang K. (2011). Partial Properties of Polyphenol Oxidase in Sour Cherry (Prunus cerasus L. CV. CAB) Pulp, World Journal of Agricultural Sciences 7 (4): 444-449, 2011.
Gawlik-Dziki U, Szymanowska U, Baraniak B. ( 2007). Characterization of
polyphenol oxidase from broccoli (Brassica oleracea var. botrytis italica)
florets. Food Chem, 105:1047-1053.
Gomez-Lopez VM. (2002). Some biochemical properties of polyphenol oxidase from two varieties of avocado. Food Chem, 77:163-169.
Gonzalez E M, Ancos B and Cano M.P. (2000), Partial characterization of peroxidase and polyphenol oxidase activities in Blackberry fruits. Journal of Agriculture and Food Chemistry, 48, 5459-5464.
Halder J, Tamuli P and Phaduri A.N. (1998), Isolation and characterization of polyphenol oxidase from Indian tea leaf (Camellia sinensis). Journal of Nutritional Biochemistry, 9, 75-80.
Jimenezatienzar M, Pedreno M, Garciacarmona F. (1991). Activation of polyphenol oxidase by Polyamines. Biochem. Int. 25, pp. 861.
Kavrayan D and Aydemir T. (2001), Partial purification and characterization of
polyphenoloxidase from peppermint (Mentha piperita). Food Chemistry, 74, 147-154.
Laveda F, Nunez-Delicado E, Garcia-Carmona F, Sanchez-Ferrer A. (2001). Proteolytic activation of latent Paraguaya peach PPO. Characterization of monophenolase activity. J. Agric. Food Chem. 49(2), pp. 1003-1008.
Lourenço E.J, Leão J.S and Neves V.A. (1990), Heat inactivation and kinetics of polyphenoloxidase from palmito (Euterpe edulis). Journal of Science Food and Agriculture, 52, 249-259.
Lourenço E.J, Neves V.A and Da Silva M.A. (1992). Polyphenoloxidase from sweet potato: Purification and properties. Journal of Agricultural and Food Chemistry, 40, 2369-2373.
Lowry O.H, A. L. Farr, Rosebrough, and R. J. Randall, “Protein measurement with the FolinPhenol Matheis, G., Whitaker, J.R. 1984, Modification of proteins by polyphenol oxidase and peroxidase and their products. J. Food Biochem. 8(3), pp. 137-162.
Okot-Kotber M, Liavoga, A, Yong K.J, Bagorogoza K. (2002). Activation of polyphenol oxidase in extracts of bran from several wheat (Triticum aestivum) cultivars using organic solvents, detergents, and chaotropes. J. Agric. Food Chem. 50(8), pp. 2410-2417.
Rapeanu G, Van Loey A, Smout C and Hendrickx M. (2006). Biochemical characterization and process stability of polyphenoloxidase extracted from Victoria grape (Vitis vinifera ssp. Sativa). Food Chemistry, 94, 253-261.
ROBB D.A. (1984). Copper Protein and Copper enzyme. In: Contie R (ed). Vol 2. pp 207-241.
CRC Press. Boca Raton, FL. Robinson DS, Eskin NAM (eds), Zawistowski J, Biliaderis CG, Eskin NAM. (1991). Oxidative Enzymes in Foods. Elsevier, New York, pp. 217-273.
Sanchez-Amat A, Solano F. (1997). A pluripotent polyphenol oxidase from the melanogenic marine alteromonas shares catalytic capabilities. Biochem .Biophys. Res. Comm. 240, pp. 787-792.
Santosh R.K, Beena P, Appu Rao A.G, GOWDA. L.R. (2006). The conformational state of polyphenol oxidase from field bean (Dolichos lablab) upon SDS and acid-pH activation. Biochem. J. 395(3), pp. 551-562.
Serradell MA, Rozenfeld PA, Martinez GA, Civello PM, Chaves AR, Anon MC. (2000). Polyphenoloxidase activity from strawberry fruit (Fragaria x ananassa, Duch., cv Selva): characterization and partial purification. J Sci Food Agric, 80:1421-1427.
Ünal MÜ, fiener A. (2006). Determination of biochemical properties of polyphenol oxidase from Emir grape (Vitis vinifera L. cv. Emir). J Sci Food Agric, 86:2374-2379.
Ünal MÜ. (2007). Properties of polyphenol oxidase from Anamur banana (Musa cavendishii).Food Chem, 100:909-913.
Van Gelder C.W, Flurkey W.H, Wichers H.J. (1997). Sequence and structural features of plant and fungal tyrosinases. Phytochemistry. 45, pp.1309-1323.
Vamos-Vigyazo, L. (1981). Polyphenol oxidase and peroxidase in fruits and
vegetables. Crit Rev Food Sci, 15:49-127.
Yang C-P, Fujita S, Kohno K, Kusubayashi A, Ashrafuzzaman MD, Hayashi N.
( 2001) .Partial purification and characterization of polyphenol oxidase from banana (Musa sapientum L) peel. J Sci Food Agric, 49:1446-1449.
Volume 1, Issue 6 - Serial Number 6
May and June 2013Pages 637-648
- Receive Date 17 August 2014
| Article View | 41,053 |
| PDF Download | 13,144 |