The Preparation of Microenvironments for Bound Enzymes by Solid Phase Peptide Synthesis
It has become evident that in the area of immobilized enzymes no single attachment system or resin support or reactor design will be optimum for all enzymes. Thus it is imperative that we develop a repertoire in these areas to allow for system designs which will either optimize the immobilized enzyme or make it more substrate selective. We sought to develop a systematic method that would allow one to specify the chemical microenvironment on the support resin onto which the enzyme is attached; thereby enabling a more systematic investigation of the influence of surface environment on enzyme structure and activity. Hornby et al., (1) Goldstein et al., (2), and Goldstein (3) have shown that the surface characteristics of the resin, i.e., its charge and hydrophobic nature, do influence the apparent kinetic parameters of immobilized enzymes. Furthermore, we (4) have shown that the spatial placement of the enzyme from the resin’s surface also affects the kinetic parameters. This led us to examine the feasibility of synthesizing peptide chains of a given chemical characteristic on the surface of a resin and attaching the enzyme to these chains. Such a method would allow comparisons of microenvironments without the interferring effects of matrix structure or enzyme distance from the surface, since these could be held constant.
KeywordsGlass Bead Immobilize Enzyme Succinic Anhydride Solid Phase Peptide Synthesis Benzyl Ester
Unable to display preview. Download preview PDF.
- 1.Hornby, W. E., Lilly, M. D. and Crook, E. M., (1968) Biochem. J. 107, 669.Google Scholar
- 4.Taylor, J. B. and Swaisgood, H. E. (1972) Biochim. Biophys. Acta, 284, 268.Google Scholar
- 5.Weetall, H. H. and Havewala, N. B. (1972) Biotechnol. Bioeng. Symp. No. 3, 241.Google Scholar
- 6.Weetall, H. H. and Hersh, L. S. (1970) Biochim. Biophys. Acta, 206, 54.Google Scholar
- 7.Inman, D. J. and Hornby, W. E. (1972) Biochem. J. 129, 255.Google Scholar
- 8.Stewart, J. M. and Young, J. D. (1969) Solid phase peptide synthesis, W. H. Freeman and Co., San Francisco.Google Scholar
- 13.Hagenmaier, H. and Frank, H. (1972) Hoppe-Seyler’s Z. Physiol. Chem. 353, 1973.Google Scholar
- 16.Rony, P. R. (1972) Biotechnol. Bioeng. Symp. No. 3, 401.Google Scholar
- 17.Gordon, A. J. and Ford, R. A. (1972) The Chemist’s Companion, Wiley-Intersic. Pub., N. Y.Google Scholar