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A bacterial isolate (O-1) which degrades different substituted benzenesulphonates was identified as a strain of Alcaligenes sp. using chemotaxonomic, DNA:DNA hybridization and serolgical methods. Its ability to mineralize orthanilic acid (2-aminobenzenesulphonate) is correlated with the presence of a 117 MDa plasmid, designated pSAH. Selective curing of pSAH resulted in the abolition of the orthanilic acid minieralization phenotype, which could be fully restored by re-establishment of pSAH. A second plasmid of 6·8 MDa, designated pME1702, with no detectable functions, is also present in Alcaligenes sp. O-1. Upon conjugation with Alcaligenes sp. O-1 as donor, the plasmid-free recipient Pseudomonas putida PaW130 acquird the ability to utilize orthanilic Alcalignes sp. O-1. Growth characteristics, the inducibility of enzyme activity (desulphonation), and expression of characteristic proteins during mineralization of orthanilic acid indicate that a plasmid-encoded cluster of genes is involed in the biodegradation of orthanilic acid. Southern blot hybridizations did not reeal any relationship between pSAH and the archetypal TOL plasmid pWW0, which also mediates degradation of substituted aromatic compounds.
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