Potencial of fermented milk for anti-adhesion therapy

在本项目开始之前，我们已经证明了皱纹乳杆菌JCM5810能够很好地附着在粘膜下模型上，其43 kDa的S层蛋白(CBSA)已经被鉴定为粘附素。与嗜酸乳杆菌1 cm1132(=ATCC4356)的CBSA和S层蛋白(SIPA)的比对表明，30个氨基酸的信号序列除1个残基外，其余均相同。两个蛋白质在C末端的同源性为77%，在N末端的同源性为30%。这些结果表明，结合区域可能位于N-末端。1998年，我们将缺失和点突变整合到CBSA基因中，并对表达的His-Tag融合蛋白与IV型胶原结合进行了鉴定。结果提示，IV型胶原结合所需的氨基酸序列为1-287个氨基酸。磅。用皱纹夜蛾JCM5810及其S层蛋白抑制产肠毒素大肠杆菌在Matrigel和固定化层粘连蛋白上的黏附，提示S层乳酸菌及其S层蛋白有可能应用于肠道感染性疾病的抗粘连治疗药物，以抑制病原菌的胶原化。

Before this project had started, we had shown that Lactobacillus crispatus JCM 5810 well adhered to submucosal model and its 43-kDa S-layer protein(CbsA) had been identified as an adhesin.In the year 1997 primary structure of CbsA was determined by DNA sequencing. Alignment of CbsA and S-layer protein(SIpA) of non-adhesive Lactobacillus acidophilus 1CM 1132(=ATCC 4356) indicated that signal sequence of 30 amino acids was identical except one residue. The identity of two proteins was 77% in C-terminal sequence and 30% in N-terminal region. These results suggested binding region might be located in the N-terminal region.In the year 1998 deletion and point mutation was incorporated into CbsA gene and the expressed His-tag fusion protein was evaluated for binding to type IV collagen. The results suggested that 1-2 87 amino acid sequence was necessary to bind type IV collagen. Lb. crispatus JCM 5810 and its S-layer protein inhibited the adherence of enterotoxigenic Escherichia coli on Matrigel and immobilized laminin, used as an submucosal model.The above results suggested S-layer-bearing lactobacillar strains and their S-layer protein have the potential to apply anti-adhesion theraputic agent for intestinal infection disease to inhibite collonization of pathogens.