1株肠道高黏附性乳杆菌的分离鉴定  

Isolation and Identification of An Adhesive Probiotic Lactobacillus Strain from Human Gastrointestinal Tract

作　　者：王斌[1] 李秋荣[1] 刘放南[1] 罗楠[1] 李幼生[1] 李宁[1] 黎介寿[1] 

机构地区：[1]南京军区南京总医院普通外科研究所全军胃肠外科重点实验室,南京210002

出　　处：《中国生物制品学杂志》2008年第6期463-466,474,共5页Chinese Journal of Biologicals

基　　金：国家重点基础研究发展规划项目(2007CB513005);京军区总医院院管课题(2006033)

摘　　要：目的从人的肠道中分离具有高黏附性能的益生乳酸菌,为其应用于临床研究提供依据。方法采用改良的LAMVAB选择性培养基,取肠黏膜活检标本进行分级分离。对分离的乳酸菌进行生理、生化鉴定和16SrRNA序列测定,并对其耐酸、耐胆盐特性、黏附性能以及抑菌特性等进行分析。结果从人肠黏膜活检标本中分离出1株乳酸菌L2,该菌能在改良的LAMVAB选择性培养基上生长,并能抵抗pH2.5的酸性环境和0.3%的高胆盐环境;体外黏附试验显示,该菌株具有较好的黏附性能,对人肠上皮样细胞Caco-2和大鼠小肠上皮细胞IEC-6的黏附指数分别达(595±125.76)/100cells和(538±65.2)/100cells,且黏附具有Ca2+依赖性。对大肠杆菌、铜绿假单胞菌等具有较强的抑制作用,对益生菌鼠李糖乳杆菌LGG和嗜酸乳杆菌L05013-12无抑制作用。该菌株为植物乳杆菌(Lactobacillus plantarum)。结论植物乳杆菌L2具有较好的黏附性能,且耐酸、耐胆盐性能以及抑菌特性均较佳,具备作为益生菌使用的潜能。Objective To isolate Lactobacillus strain with high adherence from human gastrointestinal tract for clinical use. Methods Potentially probiotic Lactobacillus strains were isolated using a modified LAMVAB selective medium from human intestinal mucosa samples collected by gastrointestinal biopsy, then subjected to physiological and biochemical identification as well as 16S rRNA sequencing, and tested for acid- and oxgall-resistances, adhesive function and antimicrobial activity. Results An Lactobacillus strain L2 was isolated, which could grow in modified LAMVAB selective medium and showed resistance to acid （pH 2.5） and 0. 3% oxgall. In vitro adhesion test showed that the adhesion indexes of L2 strain to human enteric epithelium-like cells Caco-2 and rat intestinal epithelial cells IEC-6 were （595 ± 125. 76）/100 cells and （538 ±65. 2）/100 cells respectively, and the adhesion was calcium ion-dependent. L2 strain showed strong inhibitory effect on pathogen indicators such as E. coli and Pseduomonas aeruginosa, but no inhibitory effect on probiotics such as Lactobacillus rhamnosus LGG and Lactobacillus acidophilus L050103-12. The strain was identified as Lactobacillus plantarum. Conclusion The isolated Lactobacillus plantarum L2 showed good adhesion, acid-and oxgallresistances and antimicrobial activity, and might be used as a novel probiotics.

关 键 词：乳杆菌 黏附 益生菌 分离 鉴定 

分 类 号：Q372[生物学—遗传学] Q378.2