1. Catechol was metabolized through 2-hydroxymuconic semialdehyde by cell-free extracts of benzoate-grown Azotobacter strains. Some properties of catechol 2,3 oxygenase preparations from Azotobacter vinelandii 206 are described.2 . Two different enzymatic activities able to attack 2-hydroxymuconic semialdehyde have been found in crude extracts from benzoate-grown cells ; one catalyses a hydrolytic release of formate from the semialdehyde and the other a dehydrogenation of this compound to 4-oxalocrotonate. However, the low, non-inducible levels of 2-hydroxymuconic semialdehyde hydrolase activity appear negligible for metabolic purposes and the semialdehyde seems to be dissimilated almost exclusively via 4-oxalocrotonate, by the action of a NAD+-dependent dehydrogenase, in Azotobacter strains.3, A tautomerase activity responsible for the interconversion of the enol and keto forms of 4-oxalocrotonic acid was found in extracts from benzoate-grown cells.4. 4-Oxalocrotonate was stoicheiometrically converted to CO, and 4-hydroxy-2-oxovalerate by a partially purified extract, with the transient formation of a compound that appears to be 2-oxopent-4-enoic acid. The 4-oxalocrotonate decarboxylase activity was stimulated by Mg2+ or Mn2+ ions and was inhibited by EDTA.5. Cell-free extracts from Azotobacter strains converted synthetic 4-hydroxy-2-oxovalerate to acetaldehyde and pyruvate.6. A reaction sequence, termed the 4-oxalocrotonate pathway, for the dissimilation of catechol to acetaldehyde and pyruvate by Azotobacter species is presented. All the enzymes operative in this pathway were inducible, except the 4-hydroxy-2-oxovalerate aldolase. 7. The findings described here are discussed in connection with the two previously reported meta cleavage pathways for the oxidation of catechol in Pseudomonas strains.