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

动物肠道粘附益生乳酸菌新型筛选体系的研制及应用

• 批准号: 12460117
• 负责人: SAITO Tadao
• 金额: $ 8.26万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12460117/
• 关键词: Probiotic; chicken intestinal mucin; human colon mucin; SPR; BIACORE; adhesion; rectin; cloning; プロバイオティック乳酸菌; ヒト腸管; 乳酸菌; アシドフィルス菌; 付着性; アドヘシン; 遺伝子導入; 遺伝子改変

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.

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。

项目成果

A.Matsumura et al.: "Cloning of a lectin-like protein isolated from Lactobacillus gasseri LA2 that has high adhesion activity to human intestinal mucosa"Journal of Dairy Science. 85(印刷中). (2002)

A. Matsumura 等人：“从加氏乳杆菌 LA2 中分离出的凝集素样蛋白对人肠粘膜具有高粘附活性”，《乳制品科学杂志》85（出版中）。

A. Matsumura, T. Saito, H. Kitazawa, B. Y. Kawai and T. Itoh: "Cloning of a lectin-like protein isolated from Lactobacillus gasseri LA2 that has high adhesion activity to human intestinal mucosa"Journal of Dairy Science. Vol. 85(in press). (2002)

A. Matsumura、T. Saito、H. Kitazawa、B. Y. Kawai 和 T. Itoh：“从格氏乳杆菌 LA2 中分离出的凝集素样蛋白的克隆，该蛋白对人肠粘膜具有高粘附活性”《乳品科学杂志》。