PH-DEPENDENT DECARBOXYLATION OF 2-AMINO-3-KETOBUTYRATE, THE UNSTABLE INTERMEDIATE IN THE THREONINE DEHYDROGENASE-INITIATED PATHWAY FOR THREONINE UTILIZATION

2-Amino-3-ketobutyrate can be readily formed enzymatically by the action of L-threonine dehydrogenase. A convenient assay for determining the half- life of this β-keto acid is afforded by its rapid and quantitative conversion to glycine (+ acetyl CoA), as catalyzed by 2-amino-3-ketobutyrate CoA lyase. Using this system, we have found the half-life of 2-amino-3- ketobutyrate varies with pH from 8.6 minutes at pH 5.9 to 140 minutes at pH 11.1 yielding a theoretical titration curve that predicts a pKa value of 8.15 for the α-amino group of this intermediate. These data are considered relevant to discussions pertaining to a threonine dehydrogenase/2-amino-3- ketobutyrate CoA lyase enzyme complex in the threonine utilization pathway and to mechanistic aspects of the 5-aminolevulinate synthase-catalyzed reaction where 2-amino-3-ketoadipate is involved. © 1993 Academic Press, Inc.