IN-VIVO CONSTRUCTION OF A HYBRID PATHWAY FOR METABOLISM OF 4-NITROTOLUENE IN PSEUDOMONAS-FLUORESCENS

Pseudomonas fluorescens 410PR grows on 4-nitrobenzoate but does not me tabolize 4-nitrotoluene, The TOL pWW0 Delta pm plasmid converts 4-nitr otoluene into 4-nitrobenzoate through its upper pathway, but it does n ot metabolize 4-nitrobenzoate. P, fluorescens 410PR(pWW0 Delta pm) tra nsconjugants were isolated and found to be able to grow on 4-nitrotolu ene, This phenotype was stable after growth for at least 300 generatio ns without any selective pressure, P, fluorescens 410PR(pWW0 Delta pm) converted 4-nitrotoluene into 4-nitrobenzoate via 4-nitrobenzylalcoho l and 4-nitrobenzaldehyde. 4-Nitrobenzoate was metabolized via 4-hydro xylaminobenzoate and finally yielded NH4+ and 3,4-dihydroxybenzoate, w hich was mineralized.