DETERMINATION OF THE INTRINSIC MICHAELIS CONSTANT OF IMMOBILIZED ALPHA-CHYMOTRYPSIN

The Michaelis constant of alpha-chymotrypsin, immobilized on a glutara ldehyde-activated silicate support, for N-glutaryl-L-phenylalanine-p-n itroanilide was determined and was found to be identical with that of the enzyme in solution. The influence of intraparticular diffusion was taken into account by immobilizing different amounts of enzyme, thus changing the magnitude of diffusional constraints and extrapolating ap parent Michaelis constants, determined for each amount of immobilized enzyme, to zero diffusional constraints. The possible effect of the im mobilized enzyme distribution inside the porous matrix was investigate d through numerical simulations.