Stereochemistry of the kynureninase reaction

The steric course of the replacement of the anthranilyl group of kynurenine by hydrogen was determined by conversion of (2S,3R)- and (2S,3S)-[3-3H]kynurenine into alanine with kynureninase in D2O, followed by chirality analysis of the alanine methyl group. To minimize enolization, the labeled substrates were generated in situ from the corresponding stereospecifically tritiated tryptophan species. The result, replacement in a retention mode, together with the finding that tritium from the alpha-position of the substrate is recycled and appears both at C alpha and C beta of the product suggests a single base mechanism and an active site geometry of the pyridoxal phosphate-substrate complex in which H alpha and the beta-substituent are syn oriented.