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( Sanyuan Shi ),( Liu Yang ),( Chen Yang ),( Shanshan Li ),( Hong Zhao ),( Lu Ren ),( Xiaokang Wang ),( Fuping Lu ),( Ying Li ),( Huabing Zhao ) 한국미생물생명공학회(구 한국산업미생물학회) 2021 Journal of microbiology and biotechnology Vol.31 No.2
Many bacteria metabolize aromatic compounds via catechol as a catabolic intermediate, and possess multiple genes or clusters encoding catechol-cleavage enzymes. The presence of multiple isozyme-encoding genes is a widespread phenomenon that seems to give the carrying strains a selective advantage in the natural environment over those with only a single copy. In the naphthalene-degrading strain Pseudomonas putida ND6, catechol can be converted into intermediates of the tricarboxylic acid cycle via either the ortho- or meta-cleavage pathways. In this study, we demonstrated that the catechol ortho-cleavage pathway genes (catB<sub>I</sub>C<sub>I</sub>A<sub>I</sub> and catB<sub>II</sub>C<sub>II</sub>A<sub>II</sub>) on the chromosome play an important role. The cat<sub>I</sub> and cat<sub>II</sub> operons are co-transcribed, whereas catA<sub>I</sub> and catA<sub>II</sub> are under independent transcriptional regulation. We examined the binding of regulatory proteins to promoters. In the presence of cis-cis-muconate, a well-studied inducer of the cat gene cluster, CatR<sub>I</sub> and CatR<sub>II</sub> occupy an additional downstream site, designated as the activation binding site. Notably, CatR<sub>I</sub> binds to both the cat<sub>I</sub> and cat<sub>II</sub> promoters with high affinity, while CatR<sub>II</sub> binds weakly. This is likely caused by a T to G mutation in the G/T-N11-A motif. Specifically, we found that CatR<sub>I</sub> and CatR<sub>II</sub> regulate catB<sub>I</sub>C<sub>I</sub>A<sub>I</sub> and catB<sub>II</sub>C<sub>II</sub>A<sub>II</sub> in a cooperative manner, which provides new insights into naphthalene degradation.
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