类植物乳杆菌L-ZS9生物被膜态的黏附能力及形成影响因素  

Adhesion Ability of Lactobacillus paraplantarum L-ZS9 and Influencing Factors of Its Biofilm Formation

作　　者：刘蕾[1] 武瑞赟[1,2] 李军 李平兰[1] 

机构地区：[1]中国农业大学食品科学与营养工程学院,北京食品营养与人类健康高精尖创新中心,北京100083 [2]内蒙古大学生命科学学院,内蒙古呼和浩特010021

出　　处：《食品科学》2016年第15期136-143,共8页Food Science

基　　金：国家自然科学基金面上项目(31271827,31471707);国家高技术研究发展计划(863计划)项目(2011AA100805-0-1)

摘　　要：旨在探究生物被膜态类植物乳杆菌L-ZS9的黏附能力,并确定影响其形成的主要因素。通过体外黏附实验,比较L-ZS9浮游与生物被膜两种状态下的黏附能力。结果表明,被膜态L-ZS9黏附率为浮游态的1.5倍,且黏附相关基因fbb、rop N和rrf2均上调表达,显示了被膜态的黏附优势。采用结晶紫染色法考察了培养时间、温度、营养物质、蛋白酶、群体感应自诱导信号分子(autoinducer-2,AI-2)对L-ZS9被膜形成的影响。结果显示,L-ZS9于37℃培养36 h被膜形成量最多,维持营养物质充足有利于被膜的形成,各营养物质在不同阶段发挥不同作用;低p H值和胆盐会破坏已经形成的生物被膜;胃蛋白酶和胰蛋白酶不仅抑制初始被膜的形成,且可以破坏成熟的被膜;体积分数0.5%的信号分子AI-2对被膜形成的促进作用最强,并可缓解胃蛋白酶和胰蛋白酶的作用。Adhesion of probiotics to the gastrointestinal tract is prerequisite for its beneficial function. This study aims toinvestigate the adhesion ability of biofilm-like cells of Lactobacillus paraplantarum L-ZS9 and determine factors influencingthe biofilm formation of this strain. The adhesion ability to HT-29 of biofilm cells of L-ZS9 was compared with its planktoniccells. In addition, the effects of culture time, temperature, nutrition, low pH, high bile salt, proteases and quorum sensingsignal （autoinducer-2, AI-2） on biofilm formation were investigated by crystal violet staining method. The results showedthat the adhesion ratio of biofilm cells of L-ZS9 to HT-29 was 1.5 times higher than that of its planktonic cells. The genesfbb, ropN and rrf2 were up-regulated in biofilm cells. The strain produced the largest number of biofilm cells when culturedat 37 ℃for 36 h. Sufficient nutrition promoted biofilm formation. Low pH and high bile salt destroyed the formed biofilm. Pepsinand trypsin inhibited biofilm formation and destroyed the formed biofilm as well. AI-2 （0.5%, V/V） had the strongest beneficialeffect on biofilm formation and weakened the inhibition and destruction of pepsin and trypsin. This work can lay a foundation forproducing functional foods containing biofilm probiotics with better adhesion to the gastrointestinal tract.

关 键 词：类植物乳杆菌L-ZS9 黏附 生物被膜 影响因素 信号分子AI-2 

分 类 号：Q939.99[生物学—微生物学]