Abstract
A new method for monitoring reactions catalyzed by an immobilized enzyme, cross-linked penicillin acylase aggregates (PA CLEA), is suggested. Appropriate chromogenic substrates for spectrophotometric assay of catalytic activity of immobilized enzyme were chosen and their kinetic parameters determined. Active sites in PA CLEA preparations were titrated by the suggested method; it is shown that almost all active sites are retained during immobilization. This method is characterized as highly expressive, simple, and precise, and may be used for control of PA immobilization efficiency as well as for study of operational, thermal, and pH stability immobilized enzyme preparations.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- CLEA:
-
cross-linked enzyme aggregates
- NCAPA:
-
p-nitro-o-carboxyanilide of phenylacetic acid
- NCAPG:
-
p-nitro-m-carboxyanilide of D-(-)-phenylglycine
- PA:
-
penicillin acylase
- PEG:
-
polyethylene glycol
- PMSF:
-
phenylmethylsulfonyl fluoride
References
Matsumoto, K. (1993) Bioprocess Technol., 16, 67–88.
Shewale, J. G., and Sivaraman, H. (1989) Proc. Biochem., 24, 146–154.
Bruggink, A., Roos, E. C., and de Vroom, E. (1998) Org. Proc. Res. Dev., 2, 128–133.
Youshko, M. I., Moody, H. M., Bukhanov, A. L., Boosten, W. H. J., and Švedas, V. K. (2004) Biotechnol. Bioeng., 85, 323–329.
Van Langen, L. M., de Vroom, E., van Rantwijk, F., and Sheldon, R. (1999) FEBS Lett., 456, 89–92.
Schroen, C. G., Nierstrasz, V. A., Bosma, R., Kroon, P. J., Tjeerdsma, P. S., de Vroom, E., VanderLaan, J. M., Moody, H. M., Beeftink, H. H., Janssen, A. E., and Tramper, J. (2002) Biotechnol. Bioeng., 80, 144–155.
Guranda, D. T., van Langen, L. M., van Rantwijk, F., Sheldon, R. A., and Švedas, V. K. (2001) Tetrahedron: Asymmetry, 12, 1645–1650.
Guranda, D. T., Khimiuk, A. I., van Langen, L. M., van Rantwijk, F., Sheldon, R. A., and Švedas, V. K. (2004) Tetrahedron: Asymmetry, 15, 2901–2906.
Mateo, C., Abian, O., Fernandez-Lorente, G., Pedroche, J., Fernandez-Lafuente, R., Guisan, J. M., Tam, A., and Daminati, M. (2002) Biotechnol. Progr., 18, 629–634.
Bianchi, D., Golini, P., Bortolo, R., and Cesti, P. (1996) Enzyme Microb. Technol., 18, 592–596.
Jianguo, L., Wei, C., Shuai, W., and Fan, O. (2001) Reactive, Functional Polymers, 48, 75–84.
He, J., Li, X., Evans, D. G., Duan, X., and Li, C. (2000) J. Mol. Catalysis B: Enzymatic, 11, 45–53.
Prabhune, A., and Raman, S. H. (1991) Appl. Biochem. Biotechnol., 30, 265–272.
St. Clair, N. L., and Navia, M. A. (1992) J. Am. Chem. Soc., 114, 7314–7316.
Margolin, A. L. (1996) Trends Biotechnol., 14, 223–230.
Cao, L. Q., van Rantwijk, F., and Sheldon, R. A. (2000) Org. Lett., 2, 1361–1364.
Cao, L. Q., van Langen, L. M., van Rantwijk, F., and Sheldon, R. A. (2001) J. Mol. Catalysis B: Enzymatic, 11, 665–670.
Mateo, C., Palomo, J. M., van Langen, L. M., van Rantwijk, F., and Sheldon, R. A. (2004) Biotechnol. Bioeng., 86, 273–276.
Youshko, M. I., Shamolina, T. A., Guranda, D. F., Synev, A. V., and Švedas, V. K. (1998) Biochemistry (Moscow), 63, 1104–1109.
Author information
Authors and Affiliations
Additional information
Original Russian Text © N. A. Pchelintsev, M. I. Youshko, V. K. Švedas, 2006, published in Biokhimiya, 2006, Vol. 71, No. 3, pp. 396–401.
Rights and permissions
About this article
Cite this article
Pchelintsev, N.A., Youshko, M.I. & Švedas, V.K. A new method for spectrophotometric assay of activity of cross-linked penicillin acylase aggregates. Biochemistry (Moscow) 71, 315–319 (2006). https://doi.org/10.1134/S0006297906030126
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1134/S0006297906030126