Résumé
NDM-1 (New Delhi metallo-beta-lactamase) gene encodes a metallo-beta-lactamase (MBL) with high carbapenemase activity, which makes the host bacterial strain easily dispatch the last-resort antibiotics known as carbapenems and cause global concern. Here we present the bioinformatics data showing an unexpected similarity between NDM-1 and beta-lactamase II from Erythrobacter litoralis, a marine microbial isolate. We have further expressed these two mature proteins in E. coli cells, both of which present as a monomer with a molecular mass of 25 kDa. Antimicrobial susceptibility assay reveals that they share similar substrate specificities and are sensitive to aztreonam and tigecycline. The conformational change accompanied with the zinc binding visualized by nuclear magnetic resonance, Zn(2+)-bound NDM-1, adopts at least some stable tertiary structure in contrast to the metal-free protein. Our work implies a close evolutionary relationship between antibiotic resistance genes in environmental reservoir and in the clinic, challenging the antimicrobial resistance monitoring.
Sujets)
Séquence d'acides aminés , Antibactériens , Pharmacologie , Aztréonam , Pharmacologie , Cephalosporinase , Chimie , Génétique , Métabolisme , Biologie informatique , Méthodes , Résistance bactérienne aux médicaments , Génétique , Stabilité enzymatique , Évolution moléculaire , Minocycline , Pharmacologie , Données de séquences moléculaires , Phylogenèse , Structure tertiaire des protéines , Similitude de séquences d'acides nucléiques , Sphingomonadaceae , Génétique , Tigecycline , Zinc , Pharmacologie , bêta-Lactamases , Chimie , Génétique , MétabolismeRésumé
O principal objetivo deste trabalho foi estudar a degradação de PCP por Sphingomonas chlorophenolicaem solo argiloso na presença e ausência de trigo. As concentrações de PCP foram determinadas através de Análises de Alta Performance de Cromatografia Líquida. Os efeitos tóxicos de PCP foram estudados através do monitoramento do crescimento das plantas. A biodegradação de PCP por S. chlorophenolica foi acompanhada por testes de bioluminescência de Escherichia coli HB101 pUCD607 e contagens bacterianas no solo e nas raízes. A degradação de PCP ocorreu de forma mais rápida no solo plantado e inoculado quando comparada ao solo sem plantas. Houve um aumento significativo nas populações dos organismos testados nas raízes quando comparadas com as populações presentes no solo. O monitoramento do crescimento da planta mostrou o papel protetor exercido pela S.chlorophenolica contra a toxicidade do PCP.
The main objective of this study was study the PCP degradation by Sphingomonas chlorophenolica in a loamy soil in the presence and absence of plants (Winter wheat). Measurements of PCP concentrations were carried out in a laboratory basis using High performance liquid chromatography analysis (HPLC). The toxic effect of PCP on plants was studied through the monitoring of the plant growth. The biodegradation of PCP by S. chlorophenolica in soil was assessed with a bioluminescence assay of Escherichia coli HB101 pUCD607 and bacterial analyses in roots and soil. The planted and inoculated soil showed a faster degradation when compared to the inoculated soil without plants. There was a significative increase in the populations of the organisms tested in the roots when compared to the soil. The monitoring of the plant growth showed a protective role of S. chlorophenolica against the toxicity of PCP in the loamy soil.