Food and Bioprocess Technology

, Volume 11, Issue 5, pp 1039–1049 | Cite as

The Inactivation Kinetics of Soluble and Membrane-Bound Polyphenol Oxidase in Pear during Thermal and High-Pressure Processing

  • Lei Zhou
  • Wei LiuEmail author
  • Netsanet Shiferaw TerefeEmail author
Original Paper


Polyphenol oxidase (PPO) from pear was characterized with catechol as substrate. The Michaelis constant of soluble and membrane-bound PPO were 15.6 and 23.8 mM, respectively, and their optimum pH for activity were 6.0 and 6.5, respectively. The inactivation kinetics of soluble and membrane-bound PPO during thermal (45–75 °C) and high-pressure thermal processing (600 MPa, 40–80 °C) were studied. The inactivation kinetics of pear PPO were described by a first-order model at all processing conditions. Compared to soluble PPO, membrane-bound PPO was more sensitive to thermal and high-pressure inactivation. Both soluble and membrane-bound PPO displayed higher sensitivity towards thermal inactivation at pH 3.5 (pH of pear puree made from pears dipped in citric acid prior to blending) compared to pH 4.4 (pH of non-acidified pear puree). High pressure and temperature had synergistic inactivation effects on pear PPO at pH 4.4 while slight antagonistic effects were observed at pH 3.5.


Polyphenol oxidase Pear Characterization Thermal inactivation High-pressure inactivation First-order model 



We gratefully acknowledge the support provided by Rod Smith during the high-pressure processing experiments.

Funding information

We also acknowledge the financial support of this study by the National Natural Science Foundation of China (No. 31460435).


  1. Balasubramaniam, V. M., Farkas, D., & Turek, E. J. (2008). Preserving foods through high-pressure processing. Food Technology, 62(11), 32–38.Google Scholar
  2. Bayindırli, A., Alpas, H., Bozoğlu, F., & Hızal, M. (2006). Efficiency of high pressure treatment on inactivation of pathogenic microorganisms and enzymes in apple, orange, apricot and sour cherry juices. Food Control, 17(1), 52–58. Scholar
  3. Cabanes, J., Escribano, J., Gandía-Herrero, F., García-Carmona, F., & Jiménez-Atiénzar, M. (2007). Partial purification of latent polyphenol oxidase from peach (Prunus persica L. cv. Catherina). Molecular properties and kinetic characterization of soluble and membrane-bound forms. Journal of Agricultural and Food Chemistry, 55(25), 10446–10451. Scholar
  4. Cao, X., Zhang, Y., Zhang, F., Wang, Y., Yi, J., & Liao, X. (2011). Effects of high hydrostatic pressure on enzymes, phenolic compounds, anthocyanins, polymeric color and color of strawberry pulps. Journal of the Science of Food and Agriculture, 91(5), 877–885. Scholar
  5. Chakraborty, S., Rao, P. S., & Mishra, H. N. (2014). Effect of pH on enzyme inactivation kinetics in high-pressure processed pineapple (Ananas comosus L.) puree using response surface methodology. Food and Bioprocess Technology, 7(12), 3629–3645. Scholar
  6. Chakraborty, S., Rao, P. S., & Mishra, H. N. (2015). Kinetic modeling of polyphenoloxidase and peroxidase inactivation in pineapple (Ananas comosus L.) puree during high-pressure and thermal treatments. Innovative Food Science & Emerging Technologies, 27, 57–68. Scholar
  7. Concellón, A. a., Añón, M. a. C., & Chaves, A. R. (2004). Characterization and changes in polyphenol oxidase from eggplant fruit (Solanum melongena L.) during storage at low temperature. Food Chemistry, 88(1), 17–24. Scholar
  8. Dalmadi, I., Rapeanu, G., Van Loey, A., Smout, C., & Hendrickx, M. (2006). Characterization and inactivation by thermal and pressure processing of strawberry (Fragaria ananassa) polyphenol oxidase: a kinetic study. Journal of Food Biochemistry, 30(1), 56–76. Scholar
  9. Espín, J. C., Jolivet, S., & Wichers, H. J. (1998). Inhibition of mushroom polyphenol oxidase by agaritine. Journal of Agricultural and Food Chemistry, 46(8), 2976–2980. Scholar
  10. Eisenmenger, M. J., & Reyes-De-Corcuera, J. I. (2009). High pressure enhancement of enzymes: a review. Enzyme Microbial Technology, 45(5), 331–347. Scholar
  11. Gouzi, H., Depagne, C., & Coradin, T. (2012). Kinetics and thermodynamics of the thermal inactivation of polyphenol oxidase in an aqueous extract from Agaricus bisporus. Journal of Agricultural and Food Chemistry, 60(1), 500–506. Scholar
  12. Goyeneche, R., Di Scala, K., & Roura, S. (2013). Biochemical characterization and thermal inactivation of polyphenol oxidase from radish (Raphanus sativus var. sativus). LWT - Food Science and Technology, 54(1), 57–62. Scholar
  13. Hedoux, A., Guinet, Y., & Paccou, L. (2011). Analysis of the mechanism of lysozyme pressure denaturation from Raman spectroscopy investigations, and comparison with thermal denaturation. Journal of Physical Chemistry B, 115, 6740–6748.CrossRefGoogle Scholar
  14. Ionita, E., Stanciuc, N., Aprodu, I., Rapeanu, G., & Bahrim, G. (2014). pH-induced structural changes of tyrosinase from Agaricus bisporus using fluorescence and in silico methods. Journal of the Science of Food and Agriculture, 94(11), 2338–2344. Scholar
  15. Kahn, V. (1977). Latency properties of polyphenol oxidase in two avocado cultivars differing in their rate of browning. Journal of the Science of Food and Agriculture, 28(3), 233–239. Scholar
  16. Kanade, S. R., Paul, B., Rao, A. G., & Gowda, L. R. (2006). The conformational state of polyphenol oxidase from field bean (Dolichos lablab) upon SDS and acid-pH activation. Biochemical Journal, 395(3), 551–562. Scholar
  17. Kluter, R., Nattress, D., Dunne, C., & Popper, R. (1996). Shelf life evaluation of Bartlett pears in retort pouches. Journal of Food Science, 61(6), 1297–1302. Scholar
  18. Leite, R. S. R., Gomes, E., & da Silva, R. (2007). Characterization and comparison of thermostability of purified β-glucosidases from a mesophilic Aureobasidium pullulans and a thermophilic Thermoascus aurantiacus. Process Biochemistry, 42(7), 1101–1106. Scholar
  19. Liu, F., Zhao, J. H., Gan, Z. L., & Ni, Y. Y. (2015a). Comparison of membrane-bound and soluble polyphenol oxidase in Fuji apple (Malus domestica Borkh. cv. Red Fuji). Food Chemistry, 173, 86–91. Scholar
  20. Liu, L., Cao, S., Qi, X., & Yang, Z. (2015b). The effect of pH on the activity, thermokinetics and inhibition of polyphenol oxidase from peach. Journal of Food Science and Technology, 52(11), 7465–7471. Scholar
  21. Liu, W., Liu, J., Xie, M., Liu, C., Liu, W., & Wan, J. (2009). Characterization and high-pressure microfluidization-induced activation of polyphenoloxidase from Chinese pear (Pyrus pyrifolia Nakai). Journal of Agricultural and Food Chemistry, 57(12), 5376–5380. Scholar
  22. Liu, F. G., Niu, L. Y., Li, D. J., Liu, C. Q., & Jin, B. Q. (2013a). Kinetic characterization and thermal inactivation of peroxidase in aqueous extracts from sweet corn and waxy corn. Food and Bioprocess Techonology, 6(10), 2800–2807. Scholar
  23. Liu, W., Zou, L. Q., Liu, J. P., Zhang, Z. Q., Liu, C. M., & Liang, R. H. (2013b). The effect of citric acid on the activity, thermodynamics and conformation of mushroom polyphenoloxidase. Food Chemistry, 140(1-2), 289–295. Scholar
  24. Mayer, A. M. (2006). Polyphenol oxidases in plants and fungi: going places? A review. Phytochemistry, 67(21), 2318–2331. Scholar
  25. Mishra, B. B., Gautam, S., & Sharma, A. (2012). Purification and characterisation of polyphenol oxidase (PPO) from eggplant (Solanum melongena). Food Chemistry, 134(4), 1855–1861. Scholar
  26. Siddiq, M., & Cash, J. N. (2000). Physico-chemical properties of ployphenol oxidase from d’anjou and bartlett pears (Pyrus communis L.) Journal of Food Processing and Preservation, 24(5), 353–364. Scholar
  27. Siddiq, M., Cash, J. N., Sinha, N. K., & Akhter, P. (1993). Characterization and inhibition of polyphenol oxidase from pears (Pyrus communis L. cv. Bosc and red). Journal of Food Biochemistry, 17(5), 327–337. Scholar
  28. Sulaiman, A., Soo, M. J., Farid, M., & Silva, F. V. M. (2015a). Thermosonication for polyphenoloxidase inactivation in fruits: modeling the ultrasound and thermal kinetics in pear, apple and strawberry purees at different temperatures. Journal of Food Engineering, 165, 133–140. Scholar
  29. Sulaiman, A., Soo, M. J., Yoon, M. M., Farid, M., & Silva, F. V. (2015b). Modeling the polyphenoloxidase inactivation kinetics in pear, apple and strawberry purees after high pressure processing. Journal of Food Engineering, 147, 89–94. Scholar
  30. Terefe, N. S., Buckow, R., & Versteeg, C. (2014). Quality-related enzymes in fruit and vegetable products: effects of novel food processing technologies, part 1: high-pressure processing. Critical Reviews in Food Science and Nutrition, 54(1), 24–63. Scholar
  31. Terefe, N. S., Delon, A., Buckow, R., & Versteeg, C. (2015). Blueberry polyphenol oxidase: Characterization and the kinetics of thermal and high pressure activation and inactivation. Food Chemistry, 188, 193–200. Scholar
  32. Terefe, N. S., Delon, A., & Versteeg, C. (2017). Thermal and high pressure inactivation kinetics of blueberry peroxidase. Food Chemistry, 232, 820–826. Scholar
  33. Terefe, N. S., Sheean, P., Fernando, S., & Versteeg, C. (2013). The stability of almond beta-glucosidase during combined high pressure-thermal processing: a kinetic study. Applied Microbiology and Biotechnology, 97(7), 2917–2928. Scholar
  34. Terefe, N. S., Tepper, P., Ullman, A., Knoerzer, K., & Juliano, P. (2016). High pressure thermal processing of pears: Effect on endogenous enzyme activity and related quality attributes. Innovative Food Science & Emerging Technologies, 33, 56–66. Scholar
  35. Terefe, N. S., Yang, Y. H., Knoerzer, K., Buckow, R., & Versteeg, C. (2010). High pressure and thermal inactivation kinetics of polyphenol oxidase and peroxidase in strawberry puree. Innovative Food Science & Emerging Technologies, 11(1), 52–60. Scholar
  36. Tomas-Barberan, F. A., & Espin, J. C. (2001). Phenolic compunds and related enzymes as determinants of quality in fruits and vegetables. Journal of the Science of Food and Agriculture, 81(9), 853–876. Scholar
  37. Ulker Yerliturk, F., Arslan, O., Sinan, S., Gencer, N., Ozensoy, G., & O. (2008). Characterization of polyphenoloxidase from wild pear (Pyrus elaegrifolia). Journal of Food Biochemistry, 32(3), 368–383. Scholar
  38. Weemaes, C., Ludikhuyze, L., Van den Broeck, I., Hendrickx, M., & Tobback, P. (1998a). Activity, electrophoretic characteristics and heat inactivation of polyphenoloxidases from apples, avocados, grapes, pears and plums. LWT-Food Science and Technology, 31(1), 44–49. Scholar
  39. Weemaes, C. A., Ludikhuyze, L. R., Van den Broeck, I., & Hendrickx, M. E. (1998b). Effect of pH on pressure and thermal inactivation of avocado polyphenol oxidase: a kinetic study. Journal of Agricultural and Food Chemistry, 46(7), 2785–2792. Scholar
  40. Weemaes, C. A., Ludikhuyze, L. R., Van den Broeck, I., & Hendrickx, M. E. (1998c). High pressure inactivation of polyphenoloxidases. Journal of Food Science, 63(5), 873–877. Scholar
  41. Yoruk, R., & Marshall, M. R. (2003). Physicochemical properties and function of plant polyphenol oxidase: a review. Journal of Food Biochemistry, 27(5), 361–422. Scholar
  42. Zaini, N. A., Osman, A., Hamid, A. A., Ebrahimpour, A., & Saari, N. (2013). Purification and characterization of membrane-bound polyphenoloxidase (mPPO) from snake fruit [Salacca zalacca (Gaertn.) Voss]. Food Chemistry, 136(2), 407–414. Scholar
  43. Zhou, L., Liu, W., Zou, L., Xiong, Z., Hu, X., & Chen, J. (2017). Aggregation and conformational change of mushroom (Agaricus bisporus) polyphenoloxidase subjected to thermal treatment. Food Chemistry, 214, 423–431. Scholar

Copyright information

© Crown 2018

Authors and Affiliations

  1. 1.State Key Laboratory of Food Science and TechnologyNanchang UniversityNanchangChina
  2. 2.CSIRO Agriculture and FoodWerribeeAustralia

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