Characterization of the Plasmid-Encoded Arsenic Salts Resistance Determinant from Klebsiella oxytoca D12

摘要

The arsenical resistance (ars) operon was cloned from a 67-kilobase pair (kb) plasmid, which was previously shown to be responsible for arsenic salts resistance in K, oxytoca D12. When plasmid pAE48, carrying the ars operon, was transformed into E. coli, transformed cells displayed enhanced survival in the presence of 4 mM arsenite, 50 mM arsenate, or 0.4 mM antimonite. The nucleotide sequence of the 5.6-kb fragment encoding arsenical resistance revealed five open reading frames (ORFs), which were predicted to encode polypeptides of 12.8 (arsR), 13.4 (arsD), 62.6 (arsA), 45 (arsB), and 16.7 (arsQ kilodaltons (kDa). Each ORF was preceded by a ribosome binding site. A putative promoter-like sequence was identified upstream of arsR, and a possible termination site was found downstream of arsC. When the deduced ammo acid sequences of the K. oxytoca D12 Ars proteins were compared with the amino acid sequences of the E, coli R773 Ars proteins, a significant amino acid similarity was observed (87.9% for ArsR, 89.2% for ArsD, 83.2% for ArsA, 92.6% for ArsB, and 91.3% for ArsC), suggesting an evolutionary relationship of the ars genes of E. coli plasmid R113 and K. oxytoca D12.