The pUM505 plasmid, isolated from a clinical isolate, confers resistance to ciprofloxacin (CIP) when transferred into the standard strain PAO1. CIP is an antibiotic of the quinolone family that is used to treat infections. analysis, performed to identify CIP resistance genes, revealed that the 65-amino-acid product encoded by the gene in pUM505 displays 40% amino acid identity to the aminoglycoside phosphotransferase (an enzyme that phosphorylates and inactivates aminoglycoside antibiotics). We cloned (renamed, for iprofloxacinesistance rotein,lasmid encoded) into the pUCP20 shuttle vector. The resulting recombinant plasmid, pUC-, conferred resistance to CIP on strain J53-3, suggesting that this gene encodes a protein involved in CIP resistance. Using coupled enzymatic analysis, we determined that the activity of CrpP on CIP is ATP dependent, while little activity against norfloxacin was detected, suggesting that CIP may undergo phosphorylation. Using a recombinant His-tagged CrpP protein and liquid chromatography-tandem mass spectrometry, we also showed that CIP was phosphorylated prior to its degradation. Thus, our findings demonstrate that CrpP, encoded on the pUM505 plasmid, represents a new mechanism of CIP resistance in, which involves phosphorylation of the antibiotic.
Chromium (Cr) is a highly toxic metal for microorganisms as well as plants and animal cells. Due to its widespread industrial use, Cr has become a serious pollutant in diverse environmental settings. The hexavalent form of the metal, Cr(VI), is considered a more toxic species than the relatively innocuous and less mobile Cr(III) form. The study of the interactions between microorganisms and Cr has been helpful to unravel the mechanisms allowing organisms to survive in the presence of high concentrations of Cr(VI) and to detoxify and remove the oxyanion. Various mechanisms of interactions with Cr have been identified in diverse species of bacteria and fungi, including biosorption, bioaccumulation, reduction of Cr(VI) to Cr(III), and chromate efflux. Some of these systems have been proposed as potential biotechnological tools for the bioremediation of Cr pollution using bioreactors or by in situ treatments. In this review, the interactions of microorganisms with Cr are summarised, emphasising the importance of new research avenues using advanced methodologies, including proteomic, transcriptomic, and metabolomic analyses, as well as the use of techniques based on X-ray absorption spectroscopy and electron paramagnetic resonance spectroscopy.
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