Control of polyphenol oxidase activity using a catalytic mechanism

  • D. Osuga
  • A. Van Der Schaaf
  • J. R. Whitaker


Polyphenol oxidase is one of the most deteriorative of enzymes, especially in tropical fruits, yet it is essential for color of brown tea, cocoa, coffee, raisins, some figs and prunes and plant protection. It is responsible for the unwanted black-spot formation in shrimp but is important for pigmentation of human skin. In this chapter the mechanism of action of polyphenol oxidase is discussed, including the reactions catalyzed, the kinetics with respect to the two substrates, O2 and phenol, substrate specificity and the intermediates in the reaction. Differences between monophenolase and diphenolase activities are shown mechanistically. Complete amino-acid sequences are available for polyphenol oxidases from humans and mice, Neurospora crassa (fungus), and Streptomyces glaucescens (fungus) and S. antibioticus (fungus). There is 86% strict homology in amino-acid sequence between S. glaucescens and S. antibioticus but only 24% between the Streptomyces enzymes and that from N. crassa. The human and mouse enzymes are 43% homologous; they are much larger than the fungal enzymes and have little homology with the fungal enzymes, except in the activesite histidine residues. There is also much homology between the polyphenol oxidases and hemocyanins around the active-site histidine reactions. Several methods of controlling polyphenol oxidase utilize pH, O2 exclusion, heating, ascorbic acid, sodium bisulfite, thiol compounds, kcat inactivation, competitive inhibitors and removal of phenols.


Caffeic Acid Chlorogenic Acid Histidine Residue Polyphenol Oxidase Neurospora Crassa 
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  1. Bernan, V., Filpula, D., Herber, W., Bibb, M. and Katz, E. (1985) The nucleotide sequence of the tyrosinase gene from Streptomyces antibioticus and characterization of the gene product. Gene 37: 101–110.CrossRefGoogle Scholar
  2. Bouchilloux, S., McMahill, P. and Mason, H.S. (1963) The multiple forms of mushroom tyrosinase. Purification and molecular properties of the enzymes. J. Biol. Chem. 238: 1699–1707.Google Scholar
  3. Cleland, W.W. (1963a) The kinetics of enzyme-catalyzed reactions with two or more substrates or products. I. Nomenclature and rate equations. Biochim. Biophys. Acta 67: 104–137.CrossRefGoogle Scholar
  4. Cleland, W.W. (1963b) The kinetics of enzyme-catalyzed reactions with two or more substrates or products. II. Inhibition: nomenclature and theory. Biochim. Biophys. Acta 67: 173–187.CrossRefGoogle Scholar
  5. Cleland, W.W. (1963c) The kinetics of enzyme-catalyzed reactions with two or more substrates or products. III. Prediction of initial velocity and inhibition patterns by inspection. Biochim. Biophys. Acta 67: 188–196.CrossRefGoogle Scholar
  6. Dawson, C.R. and Magee, R.J. (1955) Plant tyrosinase (polyphenol oxidase). Methods Enzymol. 2: 817–827.CrossRefGoogle Scholar
  7. Dietler, C. and Lerch, K. (1982) Reaction inactivation of tyrosinase, in Oxidases and Related Redox Systems (eds T.E. King, H.S. Mason and M. Morrison), Proceedings of the Third International Symposium, 1979. Pergamon, Oxford, pp. 305–317.Google Scholar
  8. Duckworth, H.W. and Coleman, J.E. (1970) Physicochemical and kinetic properties of mushroom tyrosinase. J. Biol. Chem. 245: 1613–1625.Google Scholar
  9. Ellman, G.L. (1959) Tissue sulfhydryl groups. Arch. Biochem. Biophys. 82: 70–77.CrossRefGoogle Scholar
  10. Embs, R.J. and Markakis, P. (1965) The mechanism of sulfite inhibition of browning caused by polyphenol oxidase. J. Food Sci. 30: 753–758.CrossRefGoogle Scholar
  11. Enzyme Nomenclature (1979) Recommendations (1978) of the Nomenclature Committee of the International Union of Biochemistry. Academic Press, New York.Google Scholar
  12. Epp, O., Colman, P., Fehlhammer, H., Bode, W., Schiffer, M. and Huber, R. (1974) Crystal and molecular structure of a dimer composed of the variable portions of the Bence-Jones protein REI. Eur.J. Biochem. 45: 513–524.CrossRefGoogle Scholar
  13. Finkle, B.J. (1964) Treatment of plant tissue to prevent browning. US Patent No. 3/126/287, 24 March.Google Scholar
  14. Finkle, B.J. and Nelson, R.F. (1963) Enzyme reactions with phenolic compounds: Effect of 0-methyl-transferase on a natural substrate of fruit polyphenoloxidase. Nature 197: 902–903.CrossRefGoogle Scholar
  15. Fling, M, Horowitz, N.H. and Heinemann, S.F. (1963) The isolation and properties of crystalline tyrosinase from Neurospora. J. Biol. Chem. 238: 2045–2053.Google Scholar
  16. Fry, D.C. and Strothkamp, K.G. (1983) Photoinactivation of Agaricus bisporus tyrosinase: Modification of the binuclear copper site. Biochemistry 22: 4949–4953.CrossRefGoogle Scholar
  17. Gaykema, W.P.J., Hol, W.G.J., Vereijken, N.M., Soeter, M.N., Bak, H.J., and Beintema, J.J. (1984) 3.2 Å Structure of the copper-containing, oxygen-carrying protein Panulirus interruptus haemocyanin. Nature 309: 23–29.CrossRefGoogle Scholar
  18. Golan-Goldhirsh, A. and Whitaker, J.R. (1984) Effect of ascorbic acid, sodium bisulfite, and thiol compounds on mushroom polyphenol oxidase. J. Agric. Food Chem. 32: 1003–1009.CrossRefGoogle Scholar
  19. Golan-Goldhirsh, A. and Whitaker, J.R. (1985) kcat inactivation of mushroom polyphenol oxidase. J. Mol. Catal. 32: 141–147.CrossRefGoogle Scholar
  20. Golan-Goldhirsh, A., Osuga, D.T., Chen, A.O. and Whitaker, J.R. (1992) Effect of ascorbic acid and copper on proteins and other polymers, in The Bioorganic Chemistry of Enzymatic Catalysis: An Homage to Myron L. Bender, (eds V.T. D’Souza and J. Feder), CRC Press, Boca Raton, pp. 61–76.Google Scholar
  21. Himmelwright, R.S., Eickman, N.C., LuBien, C.D., Lerch, K. and Solomon, E.I. (1980) Chemical and spectroscopic studies of the binuclear copper active site of Neurospora tyrosinase: comparison to hemocyanin. J. Amer. Chem. Soc. 102: 7339–7344.CrossRefGoogle Scholar
  22. Hochstein, P. and Cohen, G. (1963) The cytotoxicity of melanin precursors. Ann. NY Acad. Sci. 160: 876–886.Google Scholar
  23. Huber, M. and Lerch, K. (1988) Identification of two histidines as copper ligands in Streptomyces glaucescens tyrosinase. Biochem. 27: 5610–5615.CrossRefGoogle Scholar
  24. Huber, M. Hinterman, G. and Lerch, K. (1985) Primary Structure of tyrosinase from Streptomyces glaucescens. Biochem. 24: 6038–6044.CrossRefGoogle Scholar
  25. Hunt, M.D., Eannetta, N.T., Yu, H., Newman, S.M. and Steffens, J.C. (1993) cDNA cloning and expression of potato polyphenol oxidase. Plant Mol. Biol. 21: 59–68.CrossRefGoogle Scholar
  26. Imanaga, Y. (1955) Autoxidation of L-ascorbic acid and imidazole nucleus. II. The decomposition products of imidazole derivatives present in the autoxidation mixture. J. Biochem. 42: 669–676.Google Scholar
  27. Jolley, Jr, R.L., Nelson, R.M. and Robb, D.A. (1969) The multiple forms of mushroom tyrosinase. J. Biol. Chem. 244: 3251–3257.Google Scholar
  28. Jolley, Jr, R.L., Evans, L.H., Makino, N. and Mason, H.S. (1974) Oxytyrosinase. J. Biol. Chem. 249: 335–345.Google Scholar
  29. Kwon, B.S., Haq, A.K., Pomerantz, S.H. and Halaban, R. (1987) Isolation and sequence of a cDNA clone for human tyrosinase that maps at the mouse c-albino locus. Proc. Natl Acad. Sci. USA 84: 7473–7477.CrossRefGoogle Scholar
  30. Lerch, K. (1978) Amino-acid sequence of tyrosinase from Neurospora crassa. Proc. Natl Acad. Sci. USA 75: 3635–3639.CrossRefGoogle Scholar
  31. Lerch, K. (1981) Copper monooxygenases: tyrosinase and dopamine β-monooxygenase. Metal Ions Biol. Syst. 13: 143–186.Google Scholar
  32. Lerch, K. (1982) Primary structure of tyrosinase from Neurospora crassa. II. Complete amino-acid sequence and chemical structure of a tripeptide containing an unusual thioether. J. Biol. Chem. 257: 6414–6419.Google Scholar
  33. Lerch, K. (1983) Neurospora tyrosinase: structural, spectroscopic and catalytic properties. Mol. Cell. Biochem. 52: 125–138.CrossRefGoogle Scholar
  34. Lerch, K. (1987) Monophenol monoxygenase from Neurospora crassa. Methods Enzymol. 142: 165–169.CrossRefGoogle Scholar
  35. Lerch, K. and Ettlinger, L. (1972) Purification and characterization of a tyrosinase from Streptomyces glaucescens. Eur. J. Biochem. 31: 427–437.CrossRefGoogle Scholar
  36. Martell, A.E. and Smith, R.M. (1974). Critical Stability Constants, Vol. 1, Plenum Press, New York.Google Scholar
  37. Mason, H.S. (1965). Oxidases. Annu. Rev. Biochem. 34: 595–634.CrossRefGoogle Scholar
  38. Mason, H.S., Spencer, E. and Yamazaki, I. (1961) Identification by electron-spin resonance spectroscopy of the primary product of tyrosinase. Biochem. Biophys. Res. Commun. 4: 236–238.CrossRefGoogle Scholar
  39. McEvily, A.J., Iyengar, R. and Gross, A. (1991). Inhibition of Polyphenol Oxidase by Phenolic Compounds. The Fourth Chemical Congress of North America, 25–30 August 1991, AbstractGoogle Scholar
  40. Müller, G., Ruppert, S., Schmid, E. and Schütz, G. (1988) Functional analysis of alternatively spliced tyrosinase gene transcripts. EMBO J. 7: 2723–2730.Google Scholar
  41. Pfiffer, E. and Lercn, K. (1981) Histidine at the active site of Neurospora tyrosinase. Biochemistry 20: 6029–6035.CrossRefGoogle Scholar
  42. Pomerantz, S.H. (1963) Separation, purification and properties of two tyrosinases from hamster melanoma. J. Biol. Chem. 238: 2351–2357.Google Scholar
  43. Rivas, N.J. and Whitaker, J.R. (1973) Purification and some properties of two polyphenol oxidases from Bartlett pears. Plant Physiol. 52: 501–507.CrossRefGoogle Scholar
  44. Sapers, G.M. and Hicks, K.B. (1989) Inhibition of enzymatic browning in fruits and vegetables, in Quality Factors of Fruits and Vegetables, (ed. J.J. Jen.) ACS Symposium Series 405, pp. 29–43.Google Scholar
  45. Schartau, W., Eyerie, F., Reisinger, P., Geisert, H., Storz, H. and Linzen, B. (1983) Hemocyanin in spiders. XIX. Complete amino-acid sequence of subunit d from Eurypelma californicum hemocyanin, and comparison to chain. Hoppe-Seyler’s Z. Physiol. Chem. 364: 1383–1409.CrossRefGoogle Scholar
  46. Schneider, H.J., Drexel, R., Feldmaier, G. and Linzen. B. (1983) Hemocyanins in spiders, XVIII. Complete amino-acid sequence of subunit e from Eurypelma californicum hemocyanin. Hoppe-Seyler’s Z. Physiol Chem. 364: 1357–1381.CrossRefGoogle Scholar
  47. Schoot Uiterkamp, A.S.M. and Mason, H.S. (1973) Magnetic dipole-dipole coupled Cu(II) pairs in nitric-oxide-treated tyrosinase: A structural relationship between the active sites of tyrosinase and hemocyanin. Proc. Natl Acad. Sci. USA 70: 993–996.CrossRefGoogle Scholar
  48. Shahar, T., Henning, N., Gutfinger, T., Hareven, D. and Lifschitz, E. (1992) The tomato 66.3-kD polyphenoloxidase gene: molecular identification and developmental expression. The Plant Cell, 4: 135–147.Google Scholar
  49. Shibahara, S., Tomita, Y., Sakakura, T., Nager, C., Chaudhuri, B. and Müller, R. (1986) Cloning and expression of cDNA encoding mouse tyrosinase. Nucl. Acids Res. 14: 2413–2427.CrossRefGoogle Scholar
  50. Solomon, E.I. (1981) Binuclear copper active site. Hemocyanin, tyrosinase, and type 3 copper oxidases, in Copper Proteins, (ed. T.G. Spiro), Wiley, New York, pp. 41–108.Google Scholar
  51. Smith, R.M. and Martell, A.E. (1975) Critical Stability Constants, Vol. 3, Plenum Press, New York.CrossRefGoogle Scholar
  52. Sussman, A.S. (1961). A comparison of the properties of two forms of tyrosinase from Neurospora crassa. Arch. Biochem. Biophys. 95: 407–415.CrossRefGoogle Scholar
  53. Szent-Györgyi, A. and Vietorisz, K. (1931) Function and significance of polyphenol oxidase from potatoes. Biochem. S. 233: 236–239.Google Scholar
  54. Tainer, J.A., Getzoff, E.D., Beem, K.M., Richardson, J.S. and Richardson, D.C. (1982) Determination and analysis of the 2Å structure of copper, zinc Superoxide dismutase. J. Mol. Biol. 160: 181–217.CrossRefGoogle Scholar
  55. Uchida, K. and Kawakishi, S. (1986) Oxidative degradation of β-cyclodextrin by an ascorbic acidcopper ion system. Agric. Biol. Chem. 50: 367–373.CrossRefGoogle Scholar
  56. Wilcox, D.E., Porras, A.G., Hwang, Y.T., Lerch, K., Winkler, M.E. and Solomon, E.I. (1985) Substrate analogue binding to the coupled dinuclear copper active site in tyrosinase. J. Amer. Chem. Soc. 107: 4015–4027.CrossRefGoogle Scholar
  57. Winkler, M.E., Lerch, K. and Solomon, E.I. (1981) Competitive inhibitor binding to the binuclear copper active site in tyrosinase. J. Amer. Chem. Soc. 103: 7001–7002.CrossRefGoogle Scholar
  58. Witkop, Jr, C.J. (1984) Inherited disorders of pigmentation, in Genodermatoses: Clinics in Dermatology, (ed. R.M. Goodman), J.B. Lippincot. Philadelphia, Vol. 2, pp. 70–134.Google Scholar
  59. Wong, T.C., Luh, B.S. and Whitaker, J.R. (1971a) Isolation and characterization of polyphenol oxidases of clingstone peach. Plant Physiol. 48: 19–123.CrossRefGoogle Scholar
  60. Wong, T.C., Luh, B.S. and Whitaker, J.R. (1971b) Effect of phloroglucinol and resorcinol on the clingstone peach polyphenol oxidase-catalyzed oxidation of 4-methylcatechol. Plant Physiol. 48: 24–30.CrossRefGoogle Scholar
  61. Yasunobu, K.T. (1959) Mode of action of tyrosinase, in Pigment Cell Biology, (ed. M. Gordon), Academic Press, New York, pp. 583–608.Google Scholar

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© Springer Science+Business Media New York 1994

Authors and Affiliations

  • D. Osuga
  • A. Van Der Schaaf
  • J. R. Whitaker

There are no affiliations available

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