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Mutant strains ofPseudomonas putida (arvilla ) mt-2 which have lost the ability to grow at the expense ofm - orp -toluate (methylbenzoate) but retain the ability to grow with benzoate arise spontaneously during growth on benzoate; this genetic loss occurs to a lesser extent during growth on nonaromatic carbon sources in the presence of mitomycin C. The mutants have totally lost the activity of the enzymes of the divergentmeta pathway with the possible exception of 2-oxopent-4-enoate hydratase and 4-hydroxy-2-oxovalerate aldolase; unlike the wild type they utilize benzoate by theortho pathway. Evidence is presented that these mutants have lost a plasmid coding for the enzymes of themeta pathway, which may be transmitted back to them or into otherP. putida strains. Preliminary results from these mutants and from a mutant defective in the regulation of the plasmid-carried pathway suggest that the wild type contains two benzoate oxidase systems, one on the plasmid which is nonspecific in both its catalysis and its induction and one on the chromosome which is more specific to benzoate as substrate and is specifically induced by benzoate.

Metabolism of Benzoate and the Methylbenzoates by Pseudomonas putida ( arvilla ) mt-2: Evidence for the Existence of a TOL Plasmid