Monnappa Ajay K, Dwidar Mohammed, Seo Jeong Kon, Hur Jin-Hoe, Mitchell Robert J
1] School of Life Sciences, Ulsan National Institute of Science and Technology [2].
UNIST Central Research Facility, Ulsan National Institute of Science and Technology.
Sci Rep. 2014 Jan 22;4:3811. doi: 10.1038/srep03811.
Bdellovibrio bacteriovorus HD100 is a predatory bacterium that attacks many Gram-negative human pathogens. A serious drawback of this strain, however, is its ineffectiveness against Gram-positive strains, such as the human pathogen Staphylococcus aureus. Here we demonstrate that the extracellular proteases produced by a host-independent B. bacteriovorus (HIB) effectively degrade/inhibit the formation of S. aureus biofilms and reduce its virulence. A 10% addition of HIB supernatant caused a 75% or greater reduction in S. aureus biofilm formation as well as 75% dispersal of pre-formed biofilms. LC-MS-MS analyses identified various B. bacteriovorus proteases within the supernatant, including the serine proteases Bd2269 and Bd2321. Tests with AEBSF confirmed that serine proteases were active in the supernatant and that they impacted S. aureus biofilm formation. The supernatant also possessed a slight DNAse activity. Furthermore, treatment of planktonic S. aureus with the supernatant diminished its ability to invade MCF-10a epithelial cells by 5-fold but did not affect the MCF-10a viability. In conclusion, this study illustrates the hitherto unknown ability of B. bacteriovorus to disperse Gram-positive pathogenic biofilms and mitigate their virulence.
食菌蛭弧菌HD100是一种捕食性细菌,可攻击多种革兰氏阴性人类病原体。然而,该菌株的一个严重缺点是对革兰氏阳性菌株无效,比如人类病原体金黄色葡萄球菌。在此我们证明,一种不依赖宿主的食菌蛭弧菌(HIB)产生的细胞外蛋白酶能有效降解/抑制金黄色葡萄球菌生物膜的形成并降低其毒力。添加10%的HIB上清液可使金黄色葡萄球菌生物膜形成减少75%或更多,同时使预先形成的生物膜分散75%。液相色谱-串联质谱分析确定了上清液中的多种食菌蛭弧菌蛋白酶,包括丝氨酸蛋白酶Bd2269和Bd2321。用AEBSF进行的测试证实,丝氨酸蛋白酶在上清液中具有活性,并且它们影响金黄色葡萄球菌生物膜的形成。上清液还具有轻微的脱氧核糖核酸酶活性。此外,用上清液处理浮游的金黄色葡萄球菌可使其侵袭MCF-10a上皮细胞的能力降低5倍,但不影响MCF-10a的活力。总之,本研究揭示了食菌蛭弧菌迄今为止未知的分散革兰氏阳性致病生物膜并减轻其毒力的能力。