PNGM-1, a novel subclass B3 metallo-β-lactamase from a deep-sea sediment metagenome
PNGM-1, a novel subclass B3 metallo-β-lactamase from a deep-sea sediment metagenome
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DOI：
10.1016/j.jgar.2018.05.021
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发表时间：
2018-09-01
影响因子：
4.6
通讯作者：
Lee, Sang Hee
中科院分区：
文献类型：
--
作者：
Park, Kwang Seung;Kim, Tae Yeong;Lee, Sang Hee
目的：从爱迪生海山(距今约1万年)深海沉积物中筛选出抗菌素耐药基因(S)的功能文库。方法：从60个抗菌素耐药克隆中筛选出氨苄西林最低抑菌浓度(MIC)最高的克隆。序列分析发现一个新的金属β-内酰胺酶(MBL)基因，命名为bla(PNGm-1)。PNGM-1保留了一个锌离子结合基序(H116XH118XD120H121、H-196和H-263)，在B3MBLs亚类中保守。结果：药敏试验结果表明，携带blA基因的TOP10转化菌对青霉素类、窄谱和超广谱头孢菌素、碳青霉烯类抗生素的敏感性降低。此外，动力学分析表明，PNGM-1几乎能降解所有的β-内酰胺。结论：PNGM-1酶是来自抗生素时代之前深海沉积物的功能元基因组文库的B3MBL亚类的第一例。(C)2018年国际感染和癌症化疗学会。爱思唯尔有限公司出版。保留所有权利。
Objectives: In order to find antimicrobial resistance gene(s) pre-dating the use of antibiotics through metagenomics, functional screening of a metagenomic library from the deep-seep sediments of Edison Seamount (ca. 10 000 years old) was performed.Methods: Among 60 antimicrobial-resistant clones, a single clone with the highest minimum inhibitory concentration (MIC) for ampicillin was selected. Sequence analysis revealed a new metallo-beta-lactamase (MBL) gene, designated as bla(PNGm-1). PNGM-1 retains a zinc ion-binding motif (H116XH118XD120H121, H-196 and H-263), conserved in subclass B3 MBLs. The catalytic parameters of purified PNGM-1 and the MICs of beta-lactams for Escherichia coli TOP10 transformants harbouring the bla(PNGM-1) gene were assessed.Results: Antimicrobial susceptibility testing indicated reduced susceptibility to penicillins, narrow- and extended-spectrum cephalosporins, and carbapenems in E. coli TOP10 transformants harbouring the bla(PNGM-1) gene. In addition, kinetic analyses revealed that PNGM-1 hydrolysed almost all beta-lactams.Conclusions: The PNGM-1 enzyme is the first case of a subclass B3 MBL derived from a functional metagenomic library of a deep-sea sediment that pre-dates the antibiotic era. (C) 2018 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.