Reduction of Allergenicity of Milk Protein by Conjugation with Acidic Oligosaccharides

通过与酸性低聚糖结合降低牛奶蛋白的过敏性

基本信息

批准号：
12660113
负责人：
HATTORI Makoto
金额：
$ 2.3万
依托单位：
Tokyo University of Agriculture and Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
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/grant/KAKENHI-PROJECT-12660113/
关键词：
β-lactoglobulin protein conjugation reduction of allergenicity algmic acid oligosaccharide Maillard reaction

项目摘要

Bovine β-lactoglobulin-alginic acid oligosaccharide (β-LG-ALGO) conjugate was prepared by the Maillard reaction to reduce the allergenicity and improve the functional properties of β-LG. The molar ratio of β-LG to ALGO in the conjugates was 1 : 6. The isoelectric point of the conjugate was <4.6, which is lower than that of β-LG. Carbohydrate binding sites in β-LG were identified to be 60Lys, 77Lys, lOOLys, 138Lys and141Lys. CD spectra indicated that secondary structure of β-LG was almost maintained after conjugation with ALGO. Fluorescence studies suggested that the conformation around Trp had not changed in the conjugate and that the surface of the conjugate was covered with saccharide chain. Structural analyzes with monoclonal antibodies indicated that the conformation around 15Val29Ile and 8Lys-19-Trp in the conjugate had changed, while native structure was maintained around 125Thr-135Lys. By conjugation with ALGO, β-LG was endowed with high heat stability and improved emulsifying ability. The antiβ-LG antibody response was markedly reduced after immunization with the β-LG- ALGO conjugates in BALB/c, C57BL/6 and C3H/He mice. We determined the B and T cell epitopes of β-LG and the conjugate recognized in these mice and found that the linear epitope profiles of the β-LG- ALGO conjugate were similar to those of β-LG, while the immune response for each epitope was dramatically reduced. Masking of epitopes by ALGO was considered to be responsible for the decreased immunogenicity of the β-LG in the conjugate.

通过美拉德反应制备牛β-乳球蛋白-海藻酸寡糖(β-LG-ALGO)缀合物，以降低致敏性并改善β-LG的功能特性，缀合物中β-LG与ALGO的摩尔比为1。：6.缀合物的等电点<4.6，低于β-LG中的碳水化合物结合位点。 60Lys、77Lys、100Lys、138Lys 和 141Lys 的 CD 光谱表明，与 ALGO 缀合后，β-LG 的二级结构几乎保持不变，荧光研究表明缀合物中 Trp 周围的构象没有变化，并且缀合物的表面被覆盖。与单克隆抗体的结构引擎表明，周围的构象。结合物中的15Val29Ile和8Lys-19-Trp发生了变化，而125Thr-135Lys附近保持了天然结构。通过与ALGO结合，β-LG被赋予了高热稳定性和改善的乳化能力，抗β-LG抗体反应显着。 BALB/c、C57BL/6 和 C3H/He 中使用 β-LG-ALGO 缀合物免疫后减少我们测定了 β-LG 的 B 细胞和 T 细胞表位以及在这些小鼠中识别的缀合物，发现 β-LG-ALGO 缀合物的线性表位谱与 β-LG 的线性表位谱相似，而 β-LG 的免疫反应则不同。 ALGO 对表位的掩蔽被认为是导致缀合物中 β-LG 免疫原性降低的原因。