Simple-kinetic descriptions of alcohol dehydrogenase after immobilization on tresyl-chloride-activated agarose.
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.
D050788:Alanine Dehydrogenase; D000426:Alcohol Dehydrogenase / Q000378:metabolism*; D000594:Amino Acid Oxidoreductases / Q000378:metabolism; D004789:Enzyme Activation; D004800:Enzymes, Immobilized / Q000378:metabolism*; D007700:Kinetics; D009243:NAD / Q000378:metabolism; D010084:Oxidation-Reduction; D012685:Sepharose; D013450:Sulfones; D013816:Thermodynamics
D004800:Enzymes, Immobilized; D013450:Sulfones; D009243:NAD; C032605:2,2,2-trifluoroethanesulfonyl chloride; D012685:Sepharose; D000426:Alcohol Dehydrogenase; D000594:Amino Acid Oxidoreductases; D050788:Alanine Dehydrogenase