The production of a new selection system for probiotic lactic acid bacteria adhering to animal intestines and its application

为了筛选出对动物肠道有较强粘附性的乳酸菌，首次建立了一种新的筛选方法，并将鸡肠道粘蛋白（CIM）作为配体包被到5型传感器芯片上，以分离自鸡肠道的几种益生乳酸菌为分析物，对该方法进行了研究。通过共振单位值（RU）评价粘附性。以A型和B型两种商用生物素聚合物探针和人结肠粘蛋白（HCM）为配基，对人肠道进行了分析。本研究还成功分离了几株益生菌，成功克隆了加氏乳杆菌LA2表面层蛋白中的一种凝集素，并对其进行了核苷酸和氨基酸序列分析，证实了该凝集素与HCM联合收割机，为进一步利用新筛选的益生菌作为食品添加剂对动物健康有益。

A new selection method by using BIACORE 1000 (Pharmacia) was first developed for the mass screening for lactic acid bacteria which strongly adhere to animal intestine.For the analysis of chicken intestine, chicken intestinal mucin (CIM) was coated to the sensor chip type 5 as ligand and investigated to several probiotic lactic acid bacteria which were isolated form chicken intestine as analyte. The adhesion was evaluated by resonance unit value (RU). Several bacteria were successfully selected by this method.For the analysis of human intestine, two types of commercial biotinilated polymer probe (blood type A and B) and human colon mucin (HCM) were used for ligand. Several probiotic bacteria were also successfully isolated.One lectin in the surface layer protein of Lactobacillus gasseri LA2 which was confirmed to combine to HCM was successfully cloned and clarified the nucleotide and amino acid sequence.We demonstrated the utilization of new selected bacteria for animal health by supplementing in food as probiotic bacteria in future.