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Estimation of intrinsic kinetic constants for pore diffusion‐limited immobilized enzyme reactions
Author(s) -
Lee Gene K.,
Lesch Robert A.,
Reilly Peter J.
Publication year - 1981
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260230303
Subject(s) - michaelis–menten kinetics , thiele modulus , chemistry , diffusion , kinetics , substrate (aquarium) , reaction rate constant , immobilized enzyme , thermodynamics , enzyme kinetics , reaction rate , chromatography , catalysis , enzyme , enzyme assay , organic chemistry , active site , physics , oceanography , quantum mechanics , geology
A simple method is presented that establishes intrinsic rate parameters when slow pore diffusion of substrate limits immobilized enzyme reactions that obey Michaelis‐Menten kinetics. The Aris‐Bischoff modulus is employed. Data at high substrate concentrations, where the enzyme would be saturated in the absence of diffusion limitation, and at low substrate concentrations, where effectiveness factors are inversely proportional to reaction modulus, are used to determine maximum rate and Michaelis constant, respectively. Because Michaelis‐Menten and Langmuir‐Hinshelwood kinetics are formally identical, this method may be used to estimate intrinsic rate parameters of many heterogeneous catalysts. The technique is demonstrated using experimental data from the hydrolysis of maize dextrin with diffusion‐limited immobilized glucoamylase. This system yields a Michaelis constant of 0.14%, compared to 0.11% for soluble glucoamylase and 0.24% for immobilized glucoamylase free of diffusional effects.