title: Molecular Characterization and Computational Modelling of New Delhi Metallo-β-Lactamase-5 from an Escherichia coli Isolate (KOEC3) of Bovine Origin.
abstract: Emergence of antimicrobial resistance mediated through New Delhi metallo-β-lactamases (NDMs) is a serious therapeutic challenge. Till date, 16 different NDMs have been described. In this study, we report the molecular and structural characteristics of NDM-5 isolated from an Escherichia coli isolate (KOEC3) of bovine origin. Using PCR amplification, cloning and sequencing of full blaNDM gene, we identified the NDM type as NDM-5. Cloning of full gene in E. coli DH5α and subsequent assessment of antibiotic susceptibility of the transformed cells indicated possible role of native promoter in expression blaNDM-5. Translated amino acid sequence had two substitutions (Val88Leu and Met154Leu) compared to NDM-1. Theoretically deduced isoelectric pH of NDM-5 was 5.88 and instability index was 36.99, indicating a stable protein. From the amino acids sequence, a 3D model of the protein was computed. Analysis of the protein structure elucidated zinc coordination and also revealed a large binding cleft and flexible nature of the protein, which might be the reason for broad substrate range. Docking experiments revealed plausible binding poses for five carbapenem drugs in the vicinity of metal ions. In conclusion, results provided possible explanation for wide range of antibiotics catalyzed by NDM-5 and likely interaction modes with five carbapenem drugs.
authors: Division of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya 793103 India.
authors: D
authors: D
authors: Purkait
authors: Division of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya 793103 India.
authors: A
authors: A
authors: Ahuja
authors: Division of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya 793103 India.
authors: U
authors: U
authors: Bhattacharjee
authors: West Bengal University of Animal and Fishery Sciences, Belgachia, West Bengal 700037 India.
authors: A
authors: A
authors: Singha
authors: Division of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya 793103 India.
authors: K
authors: K
authors: Rhetso
authors: Division of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya 793103 India.
authors: T K
authors: TK
authors: Dey
authors: Division of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya 793103 India.
authors: S
authors: S
authors: Das
authors: Division of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya 793103 India.
authors: R K
authors: RK
authors: Sanjukta
authors: Division of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya 793103 India.
authors: K
authors: K
authors: Puro
authors: Division of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya 793103 India.
authors: I
authors: I
authors: Shakuntala
authors: Division of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya 793103 India.
authors: A
authors: A
authors: Sen
authors: Division of Crop Protection, ICAR Research Complex for NEH Region, Umiam, Meghalaya 793103 India.
authors: A
authors: A
authors: Banerjee
authors: Department of Microbiology, School of Life Sciences, Assam University, Silchar, Assam 788011 India.
authors: I
authors: I
authors: Sharma
authors: Pharmacokinetics and Metabolism Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors: R S
authors: RS
authors: Bhatta
authors: Nazareth Hospital, Shillong, Meghalaya 793003 India.
authors: M
authors: M
authors: Mawlong
authors: West Bengal University of Animal and Fishery Sciences, Belgachia, West Bengal 700037 India.
authors: C
authors: C
authors: Guha
authors: West Bengal University of Animal and Fishery Sciences, Belgachia, West Bengal 700037 India.
authors: N R
authors: NR
authors: Pradhan
authors: Division of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya 793103 India.
authors: S
authors: S
authors: Ghatak
keywords: Carbapenem; Cleft; Docking; Metallo-β-lactamase; Structure; blaNDM-5