Immobilized chymotrypsin on reversibly precipitable polymerized liposome

Abstract

A polymerized liposome (PLS) was prepared using a synthesized phosphatidylethanolamine with a diacetylene moiety that showed a reversibly precipitable property on addition and removal of salt. To prepare a soluble-insoluble immobilized enzyme, chymotrypsin was covalently immobilized on the outer surface of the PLS. The carbodiimide method was employed for the enzyme immobilization. Coupling was rapid and nearly complete at a weight ratio of enzyme to the PLS of < 0.12. The immobilized enzyme showed favorable activity yields for both low-and high-mol-wt substrates, i.e., 90 ±9% forN-benzoyl-L-tyrosine ethyl ester and 59 ±5% for casein up to an enzyme coupling density of 0.38 g/g-PLS. The immobilized enzyme was reusable and more stable at high temperature and long-term incubation than the native enzyme.

This is a preview of subscription content, log in to check access.

References

  1. 1.

    Charles, M., Coughlin, R. W., and Hasselberger, F. X. (1974),Biotechnol. Bioeng. 16, 1553.

    Article  CAS  Google Scholar 

  2. 2.

    Leemputten, E. V. and Horisberger, M. (1976),Biotechnol. Bioeng. 18, 587.

    Article  Google Scholar 

  3. 3.

    Margolin, A. L., Izumrudov, V. A., Svedas, V. K., and Zezin, A. B. (1982),Biotechnol. Bioeng. 24, 237.

    Article  CAS  Google Scholar 

  4. 4.

    Fujimura, M., Mori, T., and Tosa, T. (1987),Biotechnol. Bioeng. 29, 747.

    Article  CAS  Google Scholar 

  5. 5.

    Hoshino, K., Taniguchi, M., Netsu, Y., and Fujii, M. (1989),J. Chem. Eng. Jpn. 22, 54.

    Article  CAS  Google Scholar 

  6. 6.

    Okumura, K., Ikura, K., Yoshikawa, M., Sasaki, R., and Chuba, H. (1984),Agric. Biol. Chem. 48, 2435.

    CAS  Google Scholar 

  7. 7.

    Nguyen, A. L. and Luong, J. H. T. (1989),Biotechnol. Bioeng. 34, 1186.

    Article  CAS  Google Scholar 

  8. 8.

    Sun, Y., Yu, K., and Zhou, X.-Z. (dy1994), inBetter Living Through Innovative Biochem. Eng. (Proc. 3rd Asia-Pacific Biochem. Eng. Conf., Singapore, 1994), Teo, W. K., Yap, M. G. S., and Oh, S. K. W., eds., Continental Press, Singapore, pp. 613–615.

    Google Scholar 

  9. 9.

    Sun, Y., Yu, K., Jin, X. H., and Zhou, X. Z. (1995),Biotechnol. Bioeng. 47, 20.

    Article  CAS  Google Scholar 

  10. 10.

    Janolino, V. G. and Swaisgood, H. E. (1982),Biotechnol. Bioeng. 24, 1069.

    Article  CAS  Google Scholar 

  11. 11.

    Lowry, O. H., Rosenbrough, N. J., Farr, A. L., and Randall, R. J. (1951),J. Biol. Chem. 193, 265–275.

    CAS  Google Scholar 

  12. 12.

    Powers, J. D., Kilpatrick, P. K., and Carbonell, R. G. (1990),Biotechnol. Bioeng. 36, 506.

    Article  CAS  Google Scholar 

  13. 13.

    Arnon, R. and Shapira, E. (1967),Biochemistry 6, 3942.

    Article  CAS  Google Scholar 

  14. 14.

    Hub, H. H., Hupfer, B., Koch, H., and Ringsdorf, H. (1980),Angew. Chem. Int. Ed. Engl. 19, 938.

    Article  CAS  Google Scholar 

  15. 15.

    Regen, S. L., Singh, M., and Samuel, N. K. P. (1984),Biochem. Biophys. Res. Comm. 119, 646.

    Article  CAS  Google Scholar 

  16. 16.

    Kitano, H., Kato, N., Tanaka, N., and Ise, N. (1988),Biochim. Biophys. Acta. 942, 131.

    Article  CAS  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Yan Sun.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Sun, Y., Jin, X.H., Dong, X.Y. et al. Immobilized chymotrypsin on reversibly precipitable polymerized liposome. Appl Biochem Biotechnol 56, 331–339 (1996). https://doi.org/10.1007/BF02786963

Download citation

Index Entries

  • Chymotrypsin
  • immobilization
  • polymerized liposome
  • precipitation
  • stability