Enzymatic attack on immobilized substrates. 2. Diffusional limitations in the alpha-chymotrypsin-catalyzed hydrolysis of polyacrylamide-bound l-phenylalanine 4-nitroanilide.
1. The chymotrypsin-catalyzed hydrolysis of polyacrylamide-bound L-phenylalanine 4-nitroanilide was studied. As a spacer, one or two 6-aminohexanoyl residues were inserted between the matrix and ligand. 2. In the course of the enzymatic hydrolysis of polyacrylamide-bound substrates, enzyme adsorption by the gel substrates was observed. A quasi equilibrium of enzyme partitioning was reached after approximately 20-min incubation time. The enzyme adsorption could be described by the Langmuir adsorption isotherm. 3. The substrates attached via spacers to the matrix were completely hydrolyzed. 4. The initial course of the product vs time curves, as well as the dependence of the initial hydrolysis rates on enzyme concentration or substrate concentration, have been interpreted by the Nernst reaction theory. From the results obtained it has been concluded that the inital rate of the hydrolysis of polyacrylamide-bound L-phenylalanine 4-nitroanilide depends on the velocity of enzyme diffusion into the matrix.
D000178:Acrylamides; D002918:Chymotrypsin / Q000378:metabolism*; D007700:Kinetics; D008433:Mathematics; D010649:Phenylalanine / Q000031:analogs & derivatives*
D000178:Acrylamides; D010649:Phenylalanine; D002918:Chymotrypsin