Structure-function relationships in immobilized chymotrypsin catalysis
- 1 April 1983
- journal article
- research article
- Published by Wiley in Biotechnology & Bioengineering
- Vol. 25 (4), 1027-1047
- https://doi.org/10.1002/bit.260250412
Abstract
Specific activities and the amounts of active immobilized enzyme were determined for several different preparations of α‐chymotrypsin immobilized on CNBr‐activated Sepharose 4B. Electron paramagnetic resonance (EPR) spectroscopy of free and immobilized enzyme with a spin label coupled to the active site was used to probe the effects of different immobilization conditions on the immobilized enzyme active site configuration. Specific activity of active enzyme decreased and rotational correlation time of the spin label increased with increasing immobilized enzyme loading. Enzyme immobilized using an intermediate six‐carbon spacer arm exhibited greater specific activity and spin label mobility than directly coupled enzyme. The observed activity changes due to immobilization were completely consistent with corresponding active site structure alterations revealed by EPR spectroscopy.This publication has 41 references indexed in Scilit:
- Stability of immobilized α‐chymotrypsinBiotechnology & Bioengineering, 1981
- Some factors affecting the stability of glucoamylase immobilized on hornblende and on other inorganic supportsBiotechnology & Bioengineering, 1978
- The principles of enzyme stabilization I. Increase in thermostability of enzymes covalently bound to a complementary surface of a polymer support in a multipoint fashionBiochimica et Biophysica Acta (BBA) - Enzymology, 1977
- Studies on conformation of soluble and immobilized enzymes using differential scanning calorimetry. 2. Specific activity and thermal stability of enzymes bound weakly and strongly to Sepharose CL 4BBiochemistry, 1977
- Active site titration of immobilized chymotrypsin with a fluorogenic reagentFEBS Letters, 1974
- Effect of internal diffusion in heterogeneous enzyme systems: Evaluation of true kinetic parameters and substrate diffusivityJournal of Theoretical Biology, 1973
- The Denaturation by Urea and Guanidinium Chloride of Trypsin and N‐Acetylated‐Trypsin Derivatives Bound to Sephadex and AgaroseEuropean Journal of Biochemistry, 1973
- Effectiveness factors for immobilized‐enzyme reactionsThe Canadian Journal of Chemical Engineering, 1972
- Changes in conformation of insolubilized trypsin and chymotrypsin, followed by fluorescenceBiochemistry, 1971
- Chemical Fixation of Enzymes to Cyanogen Halide Activated Polysaccharide CarriersEuropean Journal of Biochemistry, 1971