Structural Requirement of Ligands for Affinity Chromatography Absorbents: Purification of Aldehyde and Xanthine Oxidases
Liver aldehyde oxidase catalyzes the oxidation of N-methyl nicotinamide to a mixture of N-methyl-4-pyridone-3-carboxamide and N-methyl-2-pyridone -5-carboxamide (1). In the past this enzyme has been purified from the livers of a number of different mammals using classical salt, organic solvent, gel and ion-exchange procedures (2). These methods suffered from being inefficient in terms of both enzyme yield and the time required to conduct purification (3). Perhaps more importantly, there is reason to believe that the homogenous enzymes so isolated were contaminated with inactive enzyme (3). In this paper, a procedure for the isolation of aldehyde oxidase by affinity chromatography will be presented. Using this method, the enzyme is recovered in high yield and with a maximum specific activity equal to or greater than that obtainable by previous methods. The procedure requires six hours to carry out and produces, as a fringe benefit, a highly purified xanthine oxidase as well.
KeywordsXanthine Oxidase Fringe Benefit Cyanogen Bromide Aldehyde Oxidase Affinity Matrix
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