Key result
Phe-228 and Leu-307 D-amino acid oxidase mutants exhibit ~71-fold higher Michaelis constants versus native enzyme.
Why the study?
The biochemical and kinetic properties of porcine kidney D-amino acid oxidase mutants Phe-228 and Leu-307 were not characterized.
Population
Recombinant Phe-228 and Leu-307 mutants of porcine kidney D-amino acid oxidase expressed in Escherichia coli
Comparison
Phe-228 and Leu-307 mutants vs native D-amino acid oxidase
Design
Preclinical expression, purification, and kinetic characterization study
Authors
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Mutant porcine DAO kinetics caution against human extrapolation; leaves open residue roles in enzyme engineering or therapeutic targeting.
Mutations at Tyr(228) and His(307) in porcine kidney D-amino acid oxidase significantly alter its kinetic properties and interaction with substrates and inhibitors.
Miyano et al. (1991) studied this question. Phe-228 and Leu-307 recombinant mutants of porcine kidney D-amino acid oxidase vs. Native/wild-type DAO was evaluated on Kinetic properties (Michaelis constants, maximum velocities, dissociation constants). Phe-228 and Leu-307 mutants of porcine kidney D-amino acid oxidase exhibited altered kinetic properties, including 71- and 10-fold higher Michaelis constants for D-alanine compared to native DAO.