文章摘要
孙梦桐;吕欣然;崔天琦;林洋;白凤翎;励建荣;.鱼类肠道中抑制哈维氏弧菌群体感应及生物膜形成的乳酸菌的筛选和鉴定[J].中国食品学报,2020,20(8):255-263
鱼类肠道中抑制哈维氏弧菌群体感应及生物膜形成的乳酸菌的筛选和鉴定
Screening and Identification of Lactic Acid Bacteria with Inhibitory Activity of Quorum Sensing and Biofilm of Vibrio harveyi from Fish Intestine
  
DOI:
中文关键词: 乳酸菌  群体感应抑制剂  哈维氏弧菌  生物膜  筛选与鉴定
英文关键词: lactic acid bacteria  quorum sensing inhibitor  Vibrio harveyi  biofilm  screening and identification
基金项目:“十三五”国家重点研发计划项目(2019YFD0901702)
作者单位
孙梦桐;吕欣然;崔天琦;林洋;白凤翎;励建荣; 渤海大学食品科学与工程学院辽宁省食品安全重点实验室生鲜农产品贮藏加工及安全控制技术国家地方联合工程研究中心
 
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中文摘要:
      从鱼类肠道来源的乳酸菌中筛选对目标菌—哈维氏弧菌群体感应及生物膜形成具有抑制作用的菌株。采用牛津杯打孔法从30株乳酸菌中筛选5株具有抗群体感应活性的菌株,其中源自鲤鱼肠道的菌株LY3-1的抑制效果较好,抑菌圈直径达14.21 mm。当菌株LY3-1粗提物质量浓度为8.0 mg/mL时,目标菌酰基高丝氨酸内酯(AHLs)信号分子的降解率为41.7%,生物膜抑制率为69.8%;当粗提物质量浓度为16.0 mg/mL时,可完全降解AHLs信号分子,生物膜抑制率达90.6%。菌株LY3-1粗提物经蛋白酶处理,群体感应抑制活性丧失,表明该粗提物中抑制群体感应的活性物质是蛋白质类物质。粗提物经40~121℃处理30 min,其群体感应抑制活性无显著性差异(P>0.05),说明其具有热稳定性;同时,该粗提物在pH 3.5~7.0范围具有群体感应抑制活性。光镜和扫描电镜结果表明:菌株LY3-1粗提物对哈维氏弧菌生物膜形态结构具有显著破坏作用。菌株LY3-1经生理生化反应和16S rRNA测序被鉴定为乳酸乳球菌,其可作为哈维氏弧菌群体感应抑制剂。本研究结果为研发革兰氏阴性菌群体感应抑制剂提供理论基础。
英文摘要:
      To screen lactic acid bacteria (LAB) with inhibitory activity of quorum sensing (QS) and biofilm formation of Vibrio harveyi from fish intestine, five strains LAB with inhibitory activity against QS of V. harveyi were screened from 30 strains LAB by the Oxford cup agar well diffusion method. Among them, strain LY3-1 isolated from Cyprinus carpio showed strong anti-QS activity and the inhibition diameter was 14.21 mm. The degradation rate of crude extracts from strain LY3-1 on acylated homoserine lactones (AHLs) signal molecular was 41.7%, and its biofilm inhibition rate was 69.8% at the mass concentration of 8.0 mg/mL. Moreover, it could completely degrade AHLs signal molecular, and its biofilm inhibition rate was 90.6% at the mass concentration of 16.0 mg/mL. The inhibitory activity of the crude extracts treated by proteases on quorum sensing of V. harveyi was lossed, indicating that the crude extracts had protein properties. After treating at 40-121 ℃ for 30 min, there was no significant difference in quorum sensing inhibitory activity of crude extracts (P>0.05), indicating that the crude extracts had heat resistance; and the crude extracts of strain LY3-1 with quorum-sensing inhibitory activity at pH 3.5-7.0. The results of light and scanning electron microscopy revealed that the crude extracts of strain LY3-1 inhibited the formation of V. harveyi biofilms. The strain LY3-1 was identified as Lactococcus lactis by physiological and biochemical characteristics and 16S rRNA gene sequencing. The study indicated that Lc. lactis could be used as a novel quorum sensing inhibitor of gram-negative bacteria. It will provide fundamental basis for the development of QS inhibitor for gram-negative bacteria quorum sensing.
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