ISOLATION AND IDENTIFICATION OF THE REACTION-PRODUCT OF ALPHA-HYDROXY-GAMMA-CARBOXYMUCONIC EPSILON-SEMIALDEHYDE DEHYDROGENASE

The reaction product of enzymic dehydrogenation of ar-hydroxy-r-carboxymuconic e-semial-dehyde (HCMS) was isolated. The analytical data (elemental analyses, IR spectra, mass spectra, proton NMR spectra, and UV spectra) showed that the product was not er-hydroxy-7-carboxymuconic acid (HCMA), the expected primary product of HCMS dehydrogenation, but a lactone of HCMA. The structure of the lactone was tentatively determined as a-pyrone-4,6-dicarboxylic acid. HCMS was converted stoichiometrically to the lactone by the purified NAD(P)-linked HCMS dehydrogenase. The lactone was actively metabolized by a cell-free extract prepared from Pseudomonas ochraceae grown on phthalic acid, suggesting that it may be a metabolic intermediate in the bacterial metabolism of protocatechuic acid. Furthermore, a method for the chemical synthesis of the lactone of HCMA is presented and some of its chemical and biochemical properties are described. © 1979, by the Japanese Biochemical Society.