Joshi Sudha, Kaur Amanjot, Sharma Prince, Harjai Kusum, Capalash Neena
Department of Biotechnology, Panjab University, Chandigarh, 160014, India,
World J Microbiol Biotechnol. 2014 Aug;30(8):2241-9. doi: 10.1007/s11274-014-1645-9. Epub 2014 Mar 27.
Pseudomonas aeruginosa possesses an arcade of both cell-associated and extracellular cytotoxic virulence factors which are regulated by a multi-component quorum sensing system. Many research studies report success of lactonase in combating the pathogenicity of P. aeruginosa but delivery of lactonase remains a challenge. The present study aims at developing a delivery vehicle for lactonase. Lactobacillus plantarum NC8 was used as host for aiiA (Bacillus thuringiensis 4A3 lactonase gene) using pSIP409 expression vector. pSIP409: aiiA construct was stably maintained in L. plantarum NC8. Co-culturing of multi-drug resistant (MDR) clinical isolates of P. aeruginosa and PAO1 with recombinant L. plantarum NC8 led to significant reduction (p < 0.001) in extracellular virulence factors like pyocyanin, protease, elastase and rhamnolipids in P. aeruginosa and also showed significant reduction in adhesion of P. aeruginosa strains to uroepithelial cells in vitro. This study shows the heterologous expression of AiiA lactonase in L. plantarum NC8. Co-culturing of lactonase expressing L. plantarum NC8 with MDR P. aeruginosa strains led to attenuation of their virulence significantly. These results underscore the potential application of recombinant L. plantarum NC8 with anti-quorum sensing properties to control infections caused by multidrug resistant P. aeruginosa.
铜绿假单胞菌拥有一系列与细胞相关和细胞外的细胞毒性毒力因子,这些因子由一个多组分群体感应系统调控。许多研究报告了内酯酶在对抗铜绿假单胞菌致病性方面的成功,但内酯酶的递送仍然是一个挑战。本研究旨在开发一种内酯酶递送载体。植物乳杆菌NC8被用作宿主,使用pSIP409表达载体来表达aiiA(苏云金芽孢杆菌4A3内酯酶基因)。pSIP409:aiiA构建体在植物乳杆菌NC8中稳定维持。将铜绿假单胞菌的多药耐药(MDR)临床分离株和PAO1与重组植物乳杆菌NC8共培养,导致铜绿假单胞菌细胞外毒力因子如绿脓菌素、蛋白酶、弹性蛋白酶和鼠李糖脂显著减少(p < 0.001),并且在体外也显示出铜绿假单胞菌菌株对尿上皮细胞的黏附显著减少。本研究展示了AiiA内酯酶在植物乳杆菌NC8中的异源表达。将表达内酯酶的植物乳杆菌NC8与多药耐药铜绿假单胞菌菌株共培养导致其毒力显著减弱。这些结果强调了具有抗群体感应特性的重组植物乳杆菌NC8在控制多药耐药铜绿假单胞菌引起的感染方面的潜在应用。