Y238, oneof thevery fewconservedresidues in theactive site ofD-amino acid oxidases (DAAO), was mutated to phe-nylalanine and serine in the enzyme from the yeastRhodo-torula gracilis. The mutated proteins are catalytically competent thus eliminating Tyr238 as an active-site acid/ base catalyst. Y238FandY238Smutants exhibit a threefold slower turnover on D-alanine as substrate, which can be attributed to a slower rate of product release relative to the wild-type enzyme (a change of the rate constants for sub-strate binding was also evident). .
