Applied Biochemistry and Biotechnology

, Volume 111, Issue 3, pp 139–152 | Cite as

Production of l-DOPA by tyrosinase immobilized on modified polystyrene

  • P. Y. Ho
  • M. S. Chiou
  • A. C. Chao


Mushroom tyrosinase was immobilized on modified polystyrene—polyaminostyrene (PSNH) and polymethylchloridestyrene (PSCL)—to produce l-DOPA from l-tyrosine. Glutaraldehyde was used as an activating agent for the PSNH to immobilize the tyrosinase and 10% (w/v) glutaraldehyde was optimal in conferring the highest specific activity (11.96 U/g) to the PSNH. Methylchloride on the PSCL was directly linked with the tyrosinase, and 1.5 mmol of Cl/g was optimal in attaining the specific activity of 17.0 U/g. The temperature and optimal acidity were, respectively, 60°C and pH 5.5 for the PSNH, and 70°C and pH 3.0 for the PSCL. In a 50-mL batch reactor working over 36 h, the l-DOPA production rate at 30°C was 1.44 mg/(L·h) for the PSNH and 2.33 mg/(L·h) for the PSCL. The production rate over 36 h was 3.86 mg/(L·h) for the PSNH at 60°C and 5.54 mg/(L·h) for the PSCL at 70°C. Both of the immobilized enzymes showed a remarkable stability with almost no change in activity after being stored wet. The operational stability study indicated a 22.4% reduction in l-DOPA production for the PSNH and an 8.63% reduction for the PSCL over seven runs (each run was for 144h at 30°C) when the immobilized enzymes were used under turnover conditions. The immobilized tyrosinase was more stable on the PSCL than on the PSNH.

Index Entries

l-DOPA tyrosine tyrosinase enzyme immobilization modified polystyrene 


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  1. 1.
    Chattopadhyay, S., Datta, S. K., and Mahato, S. B. (1994), Plant Cell Rep. 13, 519–522.CrossRefGoogle Scholar
  2. 2.
    Foor, F., Morin, N., and Bostian K. A. (1993), Appl. Environ. Microbiol. 59, 3070–3075.Google Scholar
  3. 3.
    Vilanova, E., Manjon, A., and Iborra, J. L. (1984), Biotechnol. Bioeng. 26, 1306–1312.CrossRefGoogle Scholar
  4. 4.
    Pialis, P., Hamann, M. C. J., and Saville, B. A. (1996), Biotechnol. Bioeng. 51, 141–147.CrossRefGoogle Scholar
  5. 5.
    Freire, D. G., Carvalho, G. J., and Alves, T. M. (2000), Appl. Biochem. Biotechnol. 84–86, 791–800.Google Scholar
  6. 6.
    Saville, B. A. and Seetharam, G. (2002), Enzyme Microb. Technol. 31, 747–753.CrossRefGoogle Scholar
  7. 7.
    Sánchez-Ferrer, A., Rodríguez-López, J. N., García-Cánovas, F., and García-Carmona, F. (1995), Biochimica Biophysica Acta 1247, 1–11.Google Scholar
  8. 8.
    Ge, Y. B., Wang, Y. M., Zhou, H., Wang, S. Y., Tong, Y., and Li, W. (1999), J. Biotechnol. 67, 33–40.CrossRefGoogle Scholar
  9. 9.
    Celebi, S. S. and Bahar, T. (2000), Enzyme Microb. Technol. 26, 28–33.CrossRefGoogle Scholar
  10. 10.
    Shmanai, V. V. and Bylina, G. S. (2000), React. Funct. Polymers 43, 243–251.CrossRefGoogle Scholar
  11. 11.
    Wu, C. W., Lee, J. G., and Lee, W. C. (1998), Biotechnol. Appl. Biochem. 27, 225–230.Google Scholar
  12. 12.
    Duckworth, H. W. and Coleman, J. E. (1970), J. Biol. Chem. 245, 1613–1625.Google Scholar
  13. 13.
    Hartree, E. F. (1972), Anal. Chem. 48, 422–427.Google Scholar
  14. 14.
    Lukas, A. M., Ursula, H., and Jean-Pierre, Z. (1996), Phytochemistry 42, 1511–1515.CrossRefGoogle Scholar
  15. 15.
    Bru, R., Sánchez-Ferrer, A., and García-Carmonia, F. (1988), Biotechnol. Bioeng. 34, 304–308.CrossRefGoogle Scholar
  16. 16.
    Ros, J. R., Rodríguez-López, J. N., Carlos Espín, J., Varón, R., and García-Cánovas, F. (1996), Int. J. Biochem. Cell Biol. 28, 917–923.CrossRefGoogle Scholar
  17. 17.
    García-Cánovas, F., Tudela, J., Martínez Madrid, C., Varón, R., García-Carmona, F., and Lozano, J. A. (1987), Biochimica Biophysica Acta 912, 417–423.Google Scholar
  18. 18.
    Miranda, M., Urbani, G., Di Vito, L., and Botti, D. (1979), Biochimica Biophysica Acta 585, 398–404.Google Scholar
  19. 19.
    Mateo, C., Abian, O., Fernandez-Lafuente, R., and Guisan, J. M. (2000), Enzyme Microb. Technol. 26, 509–515.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2003

Authors and Affiliations

  1. 1.Department of Chemical EngineeringNational United UniversityTaiwan, ROC

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