Physical map of the aromatic amine and m-toluate catabolic plasmid pTDN1 in Pseudomonas putida: location of a unique meta-cleavage pathway

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Abstract
Summary: Azotobacter vinelandii strain 206 degrades benzoate via the meta-cleavage pathway. In a genomic library derived from this organism a clone was obtained which carried and expressed the gene for the third enzyme in this pathway, catechol 2,3-dioxygenase (EC 1.13.11.2), on a 5.9 kb SalI restriction fragment. The structural gene was more precisely mapped on an internal 1.6 kb EcoRI fragment which, after insertion into expression vectors, directed the synthesis of a 33 kDa polypeptide. The gene showed very little or no homology with isofunctional genes derived from Pseudomonas. Comprehensive substrate specificity analysis showed significant differences between the specific activities obtained from the cloned gene product and extracts derived from Azotobacter itself.