Molecular Characterization and Computational Modelling of New Delhi Metallo-β-Lactamase-5 from an Escherichia coli Isolate (KOEC3) of Bovine Origin

被引：11

作者：

Purkait, D. ^{[1
]}

Ahuja, A. ^{[1
]}

Bhattacharjee, U. ^{[1
]}

Singha, A. ^{[2
]}

Rhetso, K. ^{[1
]}

Dey, T. K. ^{[1
]}

Das, S. ^{[1
]}

Sanjukta, R. K. ^{[1
]}

Puro, K. ^{[1
]}

Shakuntala, I. ^{[1
]}

Sen, A. ^{[1
]}

Banerjee, A. ^{[3
]}

Sharma, I. ^{[4
]}

Bhatta, R. S. ^{[5
]}

Mawlong, M. ^{[6
]}

Guha, C. ^{[2
]}

Pradhan, N. R. ^{[2
]}

Ghatak, S. ^{[1
]}

机构：

[1] ICAR Res Complex NEH Reg, Div Anim Hlth, Umiam 793103, Meghalaya, India

[2] West Bengal Univ Anim & Fishery Sci, Belgachia 700037, W Bengal, India

[3] ICAR Res Complex NEH Reg, Div Crop Protect, Umiam 793103, Meghalaya, India

[4] Assam Univ, Sch Life Sci, Dept Microbiol, Silchar 788011, Assam, India

[5] Cent Drug Res Inst, CSIR, Pharmacokinet & Metab Div, Lucknow 226001, Uttar Pradesh, India

[6] Nazareth Hosp, Shillong 793003, Meghalaya, India

来源：

INDIAN JOURNAL OF MICROBIOLOGY | 2016年 / 56卷 / 02期

关键词：

Metallo-beta-lactamase; blaNDM-5; Structure; Carbapenem; Cleft; Docking; METALLO-BETA-LACTAMASE; NDM-5; ENTEROBACTERIACEAE; PLASMID; DOCKING; PATIENT; GENE;

D O I：

10.1007/s12088-016-0569-5

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Emergence of antimicrobial resistance mediated through New Delhi metallo-beta-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 DH5 alpha 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.

引用

页码：182 / 189

页数：8

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