College of Marine Life Sciences, Ocean University of China, Qingdao, China.
J Appl Microbiol. 2012 Jun;112(6):1075-85. doi: 10.1111/j.1365-2672.2012.05291.x. Epub 2012 Apr 20.
The aim of this study was to investigate the role of membrane-bound lytic murein transglycosylase A (MltA) in a bacterial fish pathogen Edwardsiella tarda.
An mltA in-frame deletion mutant (ΔmltA) and an mltA overexpression strain (mltA(+)) of Edw. tarda were constructed through double-crossover allelic exchange and by transformation of a low-copy plasmid carrying the intact mltA into the ΔmltA mutant, respectively. Either inactivation or overexpression of MltA in Edw. tarda resulted in elevated sensitivity to β-lactam antibiotics and lower viability in oligotrophic or high osmotic environment than wild-type strain. Autolysis induced by EDTA was reduced in ΔmltA strain, while mltA(+) strain was virtually flimsy, indicating that MltA is responsible for the lysis effect. Moreover, mltA(+) strain exhibited significant increases in lipopolysaccharide (LPS) biosynthesis and virulence to zebra fish compared with wild-type strain.
The results indicated that MltA plays essential roles in β-lactam antibiotics and environmental stresses resistance, autolysis, LPS biosynthesis and pathogenicity of Edw. tarda. This is the first report that MltA has a virulence-related function in Edw. tarda.
This study provided useful information for further studies on pathogenesis of Edw. tarda.
本研究旨在探讨膜结合溶菌壁转糖基酶 A(MltA)在鱼类病原菌爱德华氏菌中的作用。
通过双交换基因同源重组构建了 Edw. tarda 的 mltA 缺失突变株(ΔmltA)和 mltA 过表达株(mltA(+)),并通过转化带有完整 mltA 的低拷贝质粒将 mltA(+)转入 ΔmltA 突变株中。Edw. tarda 中 MltA 的失活或过表达导致其对β-内酰胺类抗生素的敏感性增加,在贫营养或高渗环境中的生存能力降低。与野生型菌株相比,ΔmltA 菌株的 EDTA 诱导自溶减少,而 mltA(+)菌株几乎脆弱,表明 MltA 负责溶菌作用。此外,mltA(+)菌株的脂多糖(LPS)生物合成和对斑马鱼的毒力与野生型菌株相比显著增加。
结果表明,MltA 在 Edw. tarda 对β-内酰胺类抗生素和环境胁迫的抗性、自溶、LPS 生物合成和致病性中起重要作用。这是首次报道 MltA 在 Edw. tarda 中具有与毒力相关的功能。
本研究为进一步研究 Edw. tarda 的发病机制提供了有用的信息。