通过生物学方法检测坚强肠球菌SQ-3-2是否产生群体感应信号分子AI-2,并对检测条件进行优化。将坚强肠球菌SQ-3-2培养上清液加入到由哈维氏弧菌BB170构成的特异性报告系统中,使用多功能酶标仪化学发光模式检测荧光强度,通过与AB培养基空白对照进行荧光强度的比较得出坚强肠球菌SQ-3-2是否产生具有活性的AI-2信号分子,同时依据荧光强度的大小对加样比和酸碱度两个条件进行优化。研究结果表明,坚强肠球菌SQ-3-2培养上清液中含有AI-2信号分子,随着菌体密度的增加信号分子AI-2的浓度也随之增大,在对数期末期达到最大值。方法优化实验证明最佳检测条件为待测样品pH=7,加样比例为1∶100。实验为进一步研究坚强肠球菌SQ-3-2的Lux S系统打下基础。同时,方法优化实验为检测各类乳酸菌是否分泌AI-2信号分子提供了一定的实验基础和理论依据。

To explore that if the Enterococcus durans SQ-3-2 produces the quorum-sensing signal autoinducer-2 by utilizing a biological assay and optimize the test condition at the same time.The experiment employed a bioluminescent bacterial reporter strain called Vibrio harveyi BB170,which produces light in response to AI-2.We collected the cell-free culture fluids and added them to the V.harveyi BB170 reporter strain.The resulting light production was measured using a luminometer or scintillation counter.AI-2 activity was deduced according to the induction of luminescence of V.harveyi BB170.The fluorescence of the V.harveyi increased after adding the cell-free culture fluids of E.durans SQ-3-2 and based on the fluorescence intensity of different sample experiment condition was optimized.We found that the E.durans SQ-3-2 produces the quorum-sensing signal AI-2.With the increase of cell density,concentration of signaling molecule AI-2 increased and the maximum value was reached in logarithmic period.Optimal testing condition was sample pH=7 and sample ratio of 1∶ 100.Experiment lays the foundation to further study of function of AI-2 from E.durans SQ-3-2.At the same time,the optimized experimental conditions for detection of quorum-sensing signal AI-2 of various kinds of lactic acid bacteria is the experimental foundation.