Partial Purification and Characterisation of Polyphenol Oxidase from Two Commonly Consumed Eggplants (<i>Solanum mellongena depressum</i> and <i>Solanum gilo</i>) in Nigeria

David Morakinyo Sanni

doi:doi:10.11648/j.bmb.20160101.11

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Partial Purification and Characterisation of Polyphenol Oxidase from Two Commonly Consumed Eggplants (Solanum mellongena depressum and Solanum gilo) in Nigeria

David Morakinyo Sanni

Published in Biochemistry and Molecular Biology (Volume 1, Issue 1)

Received: 31 May 2016 Accepted: 12 June 2016 Published: 6 July 2016

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Abstract

Polyphenol oxidase (PPO) catalyzes in the presence of oxygen, the oxidation of mono- and di- phenols to o-quinones. The browning in fruits, vegetables and their processed products occurs as a result of this oxidation reaction. PPO in this study was isolated and characterized from two species of garden egg; Solanum mellongena depressum and Solanum gilo. Extracts were purified using a combination of ammonium sulphate precipitation, ion exchange chromatography on DEAE sephadex and gel filtration on Sephadex G-200 column. The effect of pH and temperature were carried out, pH and thermal stability, kinetic and substrate specificity, the effect of inhibitors and activator on the enzyme activity was investigated. About 6% protein yield for both species and a purification fold of 30.0 and 41.0 for S. depressum and S. gilo respectively was achieved. The optima pH of activity were found to be 4.0 – 4.5 and 7.0 for S. depressum and 4.0 and 8.0 for S. gilo while 4.5 and 8.0 were obtain in presence of SDS, the activity of the PPO increased in the presence of small concentration of SDS in all the pH investigated. The temperature optimum for both species was observed at 30°C. The PPO were stable at 30°C retaining about 88% and 87% of initial activity after 60minutes for S. depressum and S. gilo respectively while PPO from S. depressum was inactivated after 40min and 80°C and 70°C. A minimal remaining activity of 5% was observed at 80°C after 60min incubation time. PPO was fairly stable at pH 6.0 – 8.0 retaining a percentage remaining activity of 40.3% to 50% for S depressum and 45.5% to 52.1% for S. gilo. The PPO exhibited a marked activity towards o-diphenol and lower activities for monophenols. Ascorbic acid, EDTA and SDS inhibited enzymatic activity while the Km values using catechol, DOPA and catechin as substrate were 0.3mM, 0.095mM and 1.09mM for S. depressum and 1.9mM, 0.414mM and 0.56mM for S. gilo. PPO from S. depressum exhibited a higher enzymatic activity compared to S. gilo while S. gilo retained a fairly more stable activity.

Published in	Biochemistry and Molecular Biology (Volume 1, Issue 1)
DOI	10.11648/j.bmb.20160101.11
Page(s)	1-10
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2016. Published by Science Publishing Group

Keywords

Polyphenol Oxidase, Solanum spp., Enzyme, Activity

References

[1]	Whitaker, J. R. (1996). Polyphenol oxidase. In Food Chemistry, (O. R. Fennema, ed.) pp. 492-494, Marcel Dekker, New York.
[2]	Simpson, L., Neckelmann, N., de la Cruz, V. F., Simpson, A. M., Feagin, J. E., Jasmer, D. P., and Stuart, K. (1987). J. Biol. Chem. 262: 6182-6196.
[3]	Sanni, D. M. and Adekunle, A. A. (2012). Comparative physicochemical properties of polyphenol oxidase from white and red kolanuts (Cola nitida subspecies alba and rubra). J. Sustainable Technology, 3 (1): 9-17
[4]	Mishra, B., Gautam, S. andArun Sharma, A. (2012). Purification and characterization of polyphenol oxidase (PPO) from eggplant (Solanummelongena). Food Chemistry, 134: 1855–1861.
[5]	Cho, Y. K. and Ahn, H. K. (1999) Purification and characterization of polyphenol oxidase from potato: 1. Purification and properties. Journal of Food Biochemistry, 23: 577–592.
[6]	Rapeanu, G., Van Loey, A., Smout, C. and Hendrickx, M. (2006), Biochemical characterization and process stability of polyphenoloxidase extracted from Victoria grape (Vitisvinifera ssp. Sativa). Food Chemistry, 94: 253-261.
[7]	Lu, S. M, Tong, G. P., Long, Y. and Feng, H. (2005). Partial purification and characterization of polyphenol oxidase from fresh-cut Chinese water chestnut. J. Food Biochem. 30: 123–137.
[8]	Kader, F., Rovel, B., Girardin, M. and Metche, M. (1997). Mechanism of Browning in Fresh Highbush Blueberry Fruit corymbosum (Vaccinium L). Partial Purification and Characterisation of Blueberry Polyphenol Oxidase. J. Sci. Food Agric. 73: 513–516.
[9]	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.
[10]	Heimdal, H., Larsen, L. M. and Poll, L. (1994). Characterization of polyphenol oxidase from photosynthetic and vascular lettuce tissues (Lactucu saliva). J. Agric. Food. Chem. 42: 1428-1433.
[11]	Nozue, M., Souri, M., Arakawa, D. and Kojima, M. (1998). Purification and characterization of two isoforms of chlorogenic acid oxidase from sweet potato cells in suspension culture. J. Plant Physiol. 153: 552-557.
[12]	Oba, K., Iwatsuki, N., Uritani, I., Alvarez, A. M. and Garcia, V. V. (1992). Partial purification and characterization of polyphenol oxidase isozymes in banana bud. Biosci. Biotech. Biochem. 56: 1027-1030.
[13]	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.
[14]	Marques, L., Fleuriet, A. and Macheix, J. (1995). Characterization of multiple forms of polyphenoloxidase from apple fruit. Plant Physiol. Biochem. 33: 193-200.
[15]	Park, E. Y. and Luh, B. S. (1985). Polyphenol oxidase of kiwi fruit. Journal of Food Sci., 50: 678 - 684.
[16]	Sheptovitsky, Y. G. and Brudvig, G. W. (1996). Isolation and characterization of spinach photosystem I1 membrane-associated catalase and polyphenol oxidase. Biochemistry 35: 16255- 16263.
[17]	Laveda, F., Nunez-delicado, E., Garcia-carmona, F. and Sanchez-ferrer, A. (2001). Proteolytic activation of latent paraguaya peach PPO. Characterization of monophenolase activity. J. Agric. Food Chem. 49: 1003-1008.
[18]	Serradell, M. D. L. A., Rozenfeld, P. A., Martinez, G. A., Civello, P. M., Chaves, A. R. and Anon, M. C. (2000). Polyphenoloxidase activity from strawberry fruit (Frugariaxananassa, Duch., cv Selva) characterization and partial purification. J. Sci. Food Agric. 80: 1421- 1427.
[19]	Moore, B. M. and Flurkey, W. H. (1990). Sodium dodecyl sulfate activation of a plant polyphenoloxidase. Effect of sodium dodecyl sulfate on enzymatic and physical characteristics of purified broad bean polyphenoloxidase. J. Biol. Chem. 265: 4982-4988.
[20]	Espin, J. C. and Wichers, H. J. (1999). Activation of a latent mushroom (Agaricus bisporus) tyrosinase isoform by sodium dodecyl sulfate (SDS). Kinetic properties of the SDS-activated isoform. J. Agric. Food Chem. 47: 3518-3525.
[21]	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.
[22]	Yang, C. P., Fujita, S., Ashrafuzzaman, M., Nakamura, N., Hayashi, N. (2000). Purification and characterization of polyphenol oxidase from banana (Musa sapientumL.) pulp. J. Agric. Food Chem. 48: 2732–2735.
[23]	Robinson, S. P., Loveys, B. R. and Chacko, E. K. (1993). Polyphenol oxidase enzymes in the sap and skin of mango fruit. Aust. J. Plant Physiol. 20: 99-107.
[24]	Lee, P. M., Lee, K. and Karim, M. I. A. (1991). Biochemical studies of cocoa bean polyphenol oxidase. J. Sci. Food Agric. 55: 251-260.
[25]	Raymond, J., Rakariyatham, N. and Azanza, J. L. (1993). Purification and some properties of polyphenol oxidase from sunflower seeds. Phytochemistry, 34: 927-931.
[26]	Valero, E. and Garcia-carmona, F. (1998). pH-dependent effect of sodium chloride on latent grape polyphenol oxidase. J. Agric. Food. Chem. 46: 2447–2451.
[27]	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.
[28]	Sakiroglu, H., Kufrevioglu, O. I., Kocacaliskan, I., Oktay, M. and Onganer, Y. (1996). Purification and characterization of Dog-rose (Rosa dumlis Rechst.) polyphenol oxidase. J. Agric. Food Chem. 44: 2982-298.
[29]	Whitaker J. R. (1994). Principles of Enzymology for the Food Sciences, Second Ed., pp. 271-556, Marcel Dekker, New York.
[30]	Wesche-Ebeling, P., and Montgomery, M. W. (1990). Strawberry polyphenol oxidase: its role in anthocyanin degradation. J. Food Sci. 55: 731–734.
[31]	Lee, C. Y., Smith, N. L. and Pennesi, A. P. (1983), Polyphenol oxidase from De Chaunac grapes. Journal of Science of Food and Agriculture, 34: 987-991.
[32]	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.
[33]	López-Nicolás, J. M., Núñez-Delicado, E., Sánchez-Ferrer, Á. and García-Carmona, F. (2007): Kinetic model of apple juice enzymatic browning in the presence of cyclodextrins: the use of maltosyl-β-cyclodextrin as secondary antioxidant. Food Chemistry, 101: 1164–1171.
[34]	Mari, S., Marques, L., Breton, F., Karamanos, Y. and Macheix, J. (1998). Unfolding and refolding of active apple polyphenol oxidase. Phytochemistry, 49: 1213-1217.
[35]	Jimenez, M. and Garcia-carmona, F. (1996). The effect of sodium dodecyl sulphate on polyphenol oxidase. Phytochemistry, 42: 1503-1509.
[36]	Sanchez-ferrer, A., Bru, R. and Garcia-carmona, F. (1989). Novel procedure for extraction of a latent grape polyphenol oxidase using temperature-induced phase separation in Triton X-114. Plant Physiol. 91: 1481-1487.
[37]	Sojo, M. M., Nunez-delicado, E., Garcia-carmona, F. and Sanchez-ferrer, A. (1998). Monophenolase activity of latent banana pulp polyphenol oxidase. J. Agric. Food Chem. 46: 4931-4936.
[38]	Sanchez-ferrer, A., Laveda, F. and Garcia-carmona, F. (1993). Partial purification of soluble potato polyphenol oxidase by partitioning in aqueous two-phase system. J. Agric. Food. Chem. 41: 1219-1224.
[39]	Ferrar, P. H and Walker, J. R. L. (1996). Inhibition of diphenol oxidases; a comparative study. Journal of Food Biochemistry, 20: 15-30.

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David Morakinyo Sanni. (2016). Partial Purification and Characterisation of Polyphenol Oxidase from Two Commonly Consumed Eggplants (Solanum mellongena depressum and Solanum gilo) in Nigeria. Biochemistry and Molecular Biology, 1(1), 1-10. https://doi.org/10.11648/j.bmb.20160101.11

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David Morakinyo Sanni. Partial Purification and Characterisation of Polyphenol Oxidase from Two Commonly Consumed Eggplants (Solanum mellongena depressum and Solanum gilo) in Nigeria. Biochem. Mol. Biol. 2016, 1(1), 1-10. doi: 10.11648/j.bmb.20160101.11

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David Morakinyo Sanni. Partial Purification and Characterisation of Polyphenol Oxidase from Two Commonly Consumed Eggplants (Solanum mellongena depressum and Solanum gilo) in Nigeria. Biochem Mol Biol. 2016;1(1):1-10. doi: 10.11648/j.bmb.20160101.11

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@article{10.11648/j.bmb.20160101.11,
  author = {David Morakinyo Sanni},
  title = {Partial Purification and Characterisation of Polyphenol Oxidase from Two Commonly Consumed Eggplants (Solanum mellongena depressum and Solanum gilo) in Nigeria},
  journal = {Biochemistry and Molecular Biology},
  volume = {1},
  number = {1},
  pages = {1-10},
  doi = {10.11648/j.bmb.20160101.11},
  url = {https://doi.org/10.11648/j.bmb.20160101.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.bmb.20160101.11},
  abstract = {Polyphenol oxidase (PPO) catalyzes in the presence of oxygen, the oxidation of mono- and di- phenols to o-quinones. The browning in fruits, vegetables and their processed products occurs as a result of this oxidation reaction. PPO in this study was isolated and characterized from two species of garden egg; Solanum mellongena depressum and Solanum gilo. Extracts were purified using a combination of ammonium sulphate precipitation, ion exchange chromatography on DEAE sephadex and gel filtration on Sephadex G-200 column. The effect of pH and temperature were carried out, pH and thermal stability, kinetic and substrate specificity, the effect of inhibitors and activator on the enzyme activity was investigated. About 6% protein yield for both species and a purification fold of 30.0 and 41.0 for S. depressum and S. gilo respectively was achieved. The optima pH of activity were found to be 4.0 – 4.5 and 7.0 for S. depressum and 4.0 and 8.0 for S. gilo while 4.5 and 8.0 were obtain in presence of SDS, the activity of the PPO increased in the presence of small concentration of SDS in all the pH investigated. The temperature optimum for both species was observed at 30°C. The PPO were stable at 30°C retaining about 88% and 87% of initial activity after 60minutes for S. depressum and S. gilo respectively while PPO from S. depressum was inactivated after 40min and 80°C and 70°C. A minimal remaining activity of 5% was observed at 80°C after 60min incubation time. PPO was fairly stable at pH 6.0 – 8.0 retaining a percentage remaining activity of 40.3% to 50% for S depressum and 45.5% to 52.1% for S. gilo. The PPO exhibited a marked activity towards o-diphenol and lower activities for monophenols. Ascorbic acid, EDTA and SDS inhibited enzymatic activity while the Km values using catechol, DOPA and catechin as substrate were 0.3mM, 0.095mM and 1.09mM for S. depressum and 1.9mM, 0.414mM and 0.56mM for S. gilo. PPO from S. depressum exhibited a higher enzymatic activity compared to S. gilo while S. gilo retained a fairly more stable activity.},
 year = {2016}
}

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TY - JOUR
T1 - Partial Purification and Characterisation of Polyphenol Oxidase from Two Commonly Consumed Eggplants (Solanum mellongena depressum and Solanum gilo) in Nigeria
AU - David Morakinyo Sanni
Y1 - 2016/07/06
PY - 2016
N1 - https://doi.org/10.11648/j.bmb.20160101.11
DO - 10.11648/j.bmb.20160101.11
T2 - Biochemistry and Molecular Biology
JF - Biochemistry and Molecular Biology
JO - Biochemistry and Molecular Biology
SP - 1
EP - 10
PB - Science Publishing Group
SN - 2575-5048
UR - https://doi.org/10.11648/j.bmb.20160101.11
AB - Polyphenol oxidase (PPO) catalyzes in the presence of oxygen, the oxidation of mono- and di- phenols to o-quinones. The browning in fruits, vegetables and their processed products occurs as a result of this oxidation reaction. PPO in this study was isolated and characterized from two species of garden egg; Solanum mellongena depressum and Solanum gilo. Extracts were purified using a combination of ammonium sulphate precipitation, ion exchange chromatography on DEAE sephadex and gel filtration on Sephadex G-200 column. The effect of pH and temperature were carried out, pH and thermal stability, kinetic and substrate specificity, the effect of inhibitors and activator on the enzyme activity was investigated. About 6% protein yield for both species and a purification fold of 30.0 and 41.0 for S. depressum and S. gilo respectively was achieved. The optima pH of activity were found to be 4.0 – 4.5 and 7.0 for S. depressum and 4.0 and 8.0 for S. gilo while 4.5 and 8.0 were obtain in presence of SDS, the activity of the PPO increased in the presence of small concentration of SDS in all the pH investigated. The temperature optimum for both species was observed at 30°C. The PPO were stable at 30°C retaining about 88% and 87% of initial activity after 60minutes for S. depressum and S. gilo respectively while PPO from S. depressum was inactivated after 40min and 80°C and 70°C. A minimal remaining activity of 5% was observed at 80°C after 60min incubation time. PPO was fairly stable at pH 6.0 – 8.0 retaining a percentage remaining activity of 40.3% to 50% for S depressum and 45.5% to 52.1% for S. gilo. The PPO exhibited a marked activity towards o-diphenol and lower activities for monophenols. Ascorbic acid, EDTA and SDS inhibited enzymatic activity while the Km values using catechol, DOPA and catechin as substrate were 0.3mM, 0.095mM and 1.09mM for S. depressum and 1.9mM, 0.414mM and 0.56mM for S. gilo. PPO from S. depressum exhibited a higher enzymatic activity compared to S. gilo while S. gilo retained a fairly more stable activity.
VL - 1
IS - 1
ER -

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David Morakinyo Sanni

Department of Biochemistry, Federal University of Technology, Akure, Nigeria

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David Morakinyo Sanni. (2016). Partial Purification and Characterisation of Polyphenol Oxidase from Two Commonly Consumed Eggplants (Solanum mellongena depressum and Solanum gilo) in Nigeria. Biochemistry and Molecular Biology, 1(1), 1-10. https://doi.org/10.11648/j.bmb.20160101.11

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David Morakinyo Sanni. Partial Purification and Characterisation of Polyphenol Oxidase from Two Commonly Consumed Eggplants (Solanum mellongena depressum and Solanum gilo) in Nigeria. Biochem. Mol. Biol. 2016, 1(1), 1-10. doi: 10.11648/j.bmb.20160101.11

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David Morakinyo Sanni. Partial Purification and Characterisation of Polyphenol Oxidase from Two Commonly Consumed Eggplants (Solanum mellongena depressum and Solanum gilo) in Nigeria. Biochem Mol Biol. 2016;1(1):1-10. doi: 10.11648/j.bmb.20160101.11

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@article{10.11648/j.bmb.20160101.11,
  author = {David Morakinyo Sanni},
  title = {Partial Purification and Characterisation of Polyphenol Oxidase from Two Commonly Consumed Eggplants (Solanum mellongena depressum and Solanum gilo) in Nigeria},
  journal = {Biochemistry and Molecular Biology},
  volume = {1},
  number = {1},
  pages = {1-10},
  doi = {10.11648/j.bmb.20160101.11},
  url = {https://doi.org/10.11648/j.bmb.20160101.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.bmb.20160101.11},
  abstract = {Polyphenol oxidase (PPO) catalyzes in the presence of oxygen, the oxidation of mono- and di- phenols to o-quinones. The browning in fruits, vegetables and their processed products occurs as a result of this oxidation reaction. PPO in this study was isolated and characterized from two species of garden egg; Solanum mellongena depressum and Solanum gilo. Extracts were purified using a combination of ammonium sulphate precipitation, ion exchange chromatography on DEAE sephadex and gel filtration on Sephadex G-200 column. The effect of pH and temperature were carried out, pH and thermal stability, kinetic and substrate specificity, the effect of inhibitors and activator on the enzyme activity was investigated. About 6% protein yield for both species and a purification fold of 30.0 and 41.0 for S. depressum and S. gilo respectively was achieved. The optima pH of activity were found to be 4.0 – 4.5 and 7.0 for S. depressum and 4.0 and 8.0 for S. gilo while 4.5 and 8.0 were obtain in presence of SDS, the activity of the PPO increased in the presence of small concentration of SDS in all the pH investigated. The temperature optimum for both species was observed at 30°C. The PPO were stable at 30°C retaining about 88% and 87% of initial activity after 60minutes for S. depressum and S. gilo respectively while PPO from S. depressum was inactivated after 40min and 80°C and 70°C. A minimal remaining activity of 5% was observed at 80°C after 60min incubation time. PPO was fairly stable at pH 6.0 – 8.0 retaining a percentage remaining activity of 40.3% to 50% for S depressum and 45.5% to 52.1% for S. gilo. The PPO exhibited a marked activity towards o-diphenol and lower activities for monophenols. Ascorbic acid, EDTA and SDS inhibited enzymatic activity while the Km values using catechol, DOPA and catechin as substrate were 0.3mM, 0.095mM and 1.09mM for S. depressum and 1.9mM, 0.414mM and 0.56mM for S. gilo. PPO from S. depressum exhibited a higher enzymatic activity compared to S. gilo while S. gilo retained a fairly more stable activity.},
 year = {2016}
}

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