Characterization of D-amino acid oxidase from Trigonopsis variabilis.

D-amino acid oxidase from Trigonopsis variabilis was purified to homogeneity as a well resolved flavoprotein. Specific activity of pure enzyme was 86.6 U/mg at 30 degrees C and pH 8.5. Optimum pH for enzyme activity was 7.5 and optimum temperature was 55 degrees C. The enzyme is a non-glycosylated homodimer; the protein monomer had a M(r) of 38 +/- 2 kDa and contained one molecule of non covalently bound FAD per mole of monomer. A single molecular form with an isoelectric point of 5.1 was detected in isoelectrofocusing. The A272/A455 ratio as calculated from the absorbance spectrum was 8.4. The enzyme bound competitive inhibitors benzoate and anthranilate giving typical flavin spectral perturbations.