Department of Microbiology, Faculty of Pharmacy, Cartuja Campus, University of Granada, 18071 Granada, Spain.
Microbiol Res. 2013 Nov 7;168(9):547-54. doi: 10.1016/j.micres.2013.04.011. Epub 2013 Jun 3.
Quorum sensing (QS) systems, which depend on N-acylhomoserine lactone (AHL) signal molecules, mediate the production of virulence factors in many pathogenic microorganisms. One hundred and forty-six bacterial strains, isolated from a bivalve hatchery, were screened for their capacity to degrade five synthetic AHLs [N-butyryl-DL-homoserine lactone (C4-HSL), N-hexanoyl-DL-homoserine lactone (C6-HSL), N-octanoyl-DL-homoserine lactone (C8-HSL), N-decanoyl-DL-homoserine lactone (C10-HSL) and N-dodecanoyl-DL-homoserine lactone (C12-HSL)] using well diffusion agar-plate assays with three biosensors, Chromobacterium violaceum CV026, C. violaceum VIR07 and Agrobacterium tumefaciens NTL4 (pZLR4). The results of these assays led to our choosing four strains (PP2-67, PP2-459, PP2-644 and PP2-663) that were able to degrade all five synthetic AHLs, thus showing a wide spectrum of quorum quenching (QQ) activity. We subsequently confirmed and measured the QQ activity of the four strains by high-performance liquid chromatography plus mass-spectrometry analysis (HPLC-MS). One of the strains which showed the highest AHL-degrading activity, PP2-459, identified as being a member of the genus Thalassomonas was chosen for further study. Finally, using thin-layer chromatography (TLC), we went on to confirm this strain's capacity to degrade the AHLs produced by other non-pathogenic and pathogenic bacteria not taxonomically related.
群体感应(QS)系统依赖于 N-酰基高丝氨酸内酯(AHL)信号分子,介导许多致病性微生物中毒力因子的产生。从一个双壳贝类孵化场分离的 146 株细菌菌株,通过使用三种生物传感器(紫色杆菌 CV026、紫色杆菌 VIR07 和根癌农杆菌 NTL4(pZLR4))的琼脂扩散平板试验,筛选其降解五种合成 AHLs[N-丁酰基-DL-高丝氨酸内酯(C4-HSL)、N-己酰基-DL-高丝氨酸内酯(C6-HSL)、N-辛酰基-DL-高丝氨酸内酯(C8-HSL)、N-癸酰基-DL-高丝氨酸内酯(C10-HSL)和 N-十二酰基-DL-高丝氨酸内酯(C12-HSL)]的能力。这些试验的结果导致我们选择了四株(PP2-67、PP2-459、PP2-644 和 PP2-663)能够降解所有五种合成 AHLs 的菌株,从而显示出广泛的群体感应(QQ)活性。随后,我们通过高效液相色谱加质谱分析(HPLC-MS)确认和测量了这四株菌的 QQ 活性。其中一株表现出最高 AHL 降解活性的菌株 PP2-459 被鉴定为属于海洋单胞菌属的成员,被选为进一步研究。最后,我们通过薄层层析(TLC)进一步证实了该菌株能够降解与非致病性和致病性细菌相关的其他非分类相关细菌产生的 AHLs。