Open AccessSimple‐kinetic descriptions of alcohol dehydrogenase after immobilization on tresyl‐chloride‐activated agarose
Author(s) -
BILLE Vincent,
REMACLE José
Publication year - 1986
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1986.tb09977.x
Subject(s) - agarose , alcohol dehydrogenase , chemistry , cofactor , immobilized enzyme , michaelis–menten kinetics , enzyme , alcohol , dehydrogenase , chloride , yeast , enzyme assay , chromatography , biochemistry , organic chemistry
Yeast alcohol dehydrogenase was successfully immobilized on tresyl‐chloride‐activated agarose; the optimized conditions allowed an enzyme activity recovery of over 90%. Comparison of free and immobilized enzyme properties showed an unchanged intrinsic activation energy of the reaction and a shift of optimum activity to a higher pH medium after immobilization. Comparison of the kinetic parameters for both substrates of the reaction showed that the Michaelis‐Menten model could not take into consideration all the constraints induced by the immobilization on the enzyme properties but that the Theorell‐Chance model was more appropriate. These results are discussed taking into consideration the factors affecting the immobilized enzyme. Finally, we discuss the possibilities of cofactor regeneration with this immobilized alcohol dehydrogenase.