Degradation of chloro- and methyl-substituted benzoic acids by a genetically modified microorganism

Degradation of 3-chlorobenzoic acid (3CB), 4-chlorobenzoic acid (4CB), and 4-methylbenzoic acid (4MB) as single substrates (carbon sources) and as a substrate mixture were studied in batch and continuous culture using the genetically modified microorganism Pseudomonas sp. B13 FR1 SN45P. The strain was able to mineralize the single compounds as well as the substrate mixture completely. Conversion of the three compounds in the substrate mixture proceeded simultaneously. Maximum specific substrate conversion rates were calculated to be 0.9 g g(-1) h(-1) for 3 CB and 4CB and 1.1 g g(-1) h(-1) for 4MB. Mass balances indicated the transient accumulation of pathway intermediates during batch cultivations. Hence, the rate limiting step in the degradative pathway is not the initial microbial attack of the original substrate or its transport through the cell membrane. Degradation rates on 3CB were comparable to those of the parent strain Pseudomonas sp. B13. The stability of the degradation pathways of strain Pseudomonas sp. B13 FR1 SN45P could be demonstrated in a continuous cultivation over 3.5 months (734 generation times) on 3CB, 4MB, and 4CB, which were used as single carbon sources one after the other.