Department of Infectious Diseases, the First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China.
J Antibiot (Tokyo). 2011 Apr;64(4):317-20. doi: 10.1038/ja.2011.1. Epub 2011 Feb 9.
The purpose of this study was to investigate the phenotypic and molecular characterization of a novel plasmid-mediated AmpC-type β-lactamase in Klebsiella pneumoniae E701 isolated from Anhui province in China. In comparison with the ACT-1, sequence analysis revealed that there were 43 point mutations in the coding gene, and 10 of which led to amino-acid substitution. Resistance could be transferred by conjugation or transformation with plasmid DNA into E. coli JM109, which was due to the production of a β-lactamase with an isoelectric point of 8.4 named ACT-6. Cloning, expression, purification and kinetics were carried out to study the characterization of the novel AmpC-type β-lactamase. The results of MIC determinations and substrate profiles showed there was no significant difference in the activities of the novel enzyme and ACT-1. Moreover, the class 1 integron and the whole open reading frame of the novel AmpC-type β-lactamase from K.pneumoniae E701 were detectable in the same size plasmid. This is the first report on the emergence of the novel ACT-6 type β-lactamases in K. pneumoniae.
本研究旨在探究从中国安徽省分离出的肺炎克雷伯菌 E701 中一种新型质粒介导的 AmpC 型β-内酰胺酶的表型和分子特征。与 ACT-1 相比,序列分析显示编码基因有 43 个点突变,其中 10 个导致氨基酸取代。通过接合或转化将质粒 DNA 转入 E. coli JM109 可转移耐药性,这是由于产生了一种等电点为 8.4 的β-内酰胺酶,命名为 ACT-6。通过克隆、表达、纯化和动力学研究对新型 AmpC 型β-内酰胺酶的特性进行了研究。MIC 测定和底物谱结果表明,新型酶和 ACT-1 的活性没有显著差异。此外,肺炎克雷伯菌 E701 中的新型 AmpC 型β-内酰胺酶的 1 类整合子和完整开放阅读框可在相同大小的质粒中检测到。这是首次报道肺炎克雷伯菌中出现新型 ACT-6 型β-内酰胺酶。