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.
Similar content being viewed by others
References
Charles, M., Coughlin, R. W., and Hasselberger, F. X. (1974),Biotechnol. Bioeng. 16, 1553.
Leemputten, E. V. and Horisberger, M. (1976),Biotechnol. Bioeng. 18, 587.
Margolin, A. L., Izumrudov, V. A., Svedas, V. K., and Zezin, A. B. (1982),Biotechnol. Bioeng. 24, 237.
Fujimura, M., Mori, T., and Tosa, T. (1987),Biotechnol. Bioeng. 29, 747.
Hoshino, K., Taniguchi, M., Netsu, Y., and Fujii, M. (1989),J. Chem. Eng. Jpn. 22, 54.
Okumura, K., Ikura, K., Yoshikawa, M., Sasaki, R., and Chuba, H. (1984),Agric. Biol. Chem. 48, 2435.
Nguyen, A. L. and Luong, J. H. T. (1989),Biotechnol. Bioeng. 34, 1186.
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.
Sun, Y., Yu, K., Jin, X. H., and Zhou, X. Z. (1995),Biotechnol. Bioeng. 47, 20.
Janolino, V. G. and Swaisgood, H. E. (1982),Biotechnol. Bioeng. 24, 1069.
Lowry, O. H., Rosenbrough, N. J., Farr, A. L., and Randall, R. J. (1951),J. Biol. Chem. 193, 265–275.
Powers, J. D., Kilpatrick, P. K., and Carbonell, R. G. (1990),Biotechnol. Bioeng. 36, 506.
Arnon, R. and Shapira, E. (1967),Biochemistry 6, 3942.
Hub, H. H., Hupfer, B., Koch, H., and Ringsdorf, H. (1980),Angew. Chem. Int. Ed. Engl. 19, 938.
Regen, S. L., Singh, M., and Samuel, N. K. P. (1984),Biochem. Biophys. Res. Comm. 119, 646.
Kitano, H., Kato, N., Tanaka, N., and Ise, N. (1988),Biochim. Biophys. Acta. 942, 131.
Author information
Authors and Affiliations
Rights 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
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02786963