Immunological comparison of enzymes of the beta-ketoadipate pathway.

beta-Carboxy-cis,cis-muconate lactonizing enzyme and gamma-carboxymuconolactone decarboxylase catalyze sequential reactions in the beta-ketoadipate pathway; the subunit sizes of the enzymes from Pseudomonas putida, biotype A, are 40 000 and 13 000, respectively. The cross reaction of antisera prepared against the enzymes was tested with the isofunctional enzymes formed by representatives of other bacterial species. Despite the differences in the subunit sizes of the enzymes, the antisera revealed the same general pattern: cross reaction was observed with the corresponding enzymes formed by other strains in the fluorescent Pseudomonas RNA homology group I and generally was not observed with enzymes from other Pseudomonas species or from other bacterial genera. Exceptions were provided by representatives of Pseudomonas cepacia. Members of this species are classified outside the fluorescent Pseudomonas RNA homology group. Nevertheless, the gamma-carboxymuconolactone decarboxylases from these organisms formed precipitin bands with antisera prepared against the corresponding enzyme from P.putida, biotype A; the lactonizing enzymes from the two species did not appear to cross react. Immunodiffusion experiments with gamma-carboxymuconolactone decarboxylase indicated that a common set of antigenic determinants for the enzyme is conserved among strains that have been classified together by other criteria; the relative immunological distances of the decarboxylases of each taxon from the reference P.putida, biotype A, enzyme were indicated by spurring patterns on Ouchterlony plates. These results suggested that the interspecific transfer of the structural gene for the enzyme is not a common event in Pseudomonas.