Hataishi Yui, Tanaka Aya, Ishizuka Misaki, Mizobuchi Hibine, Yoshida Tadashi, Hattori Makoto
Department of Applied Biological Science, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-Cho, Fuchu City, Tokyo 183-8509 Japan.
Cytotechnology. 2025 Feb;77(1):6. doi: 10.1007/s10616-024-00664-9. Epub 2024 Nov 21.
β-Conglycinin was conjugated with carboxymethyl cellulose (CMC) by using water-soluble carbodiimide to improve its function. Two kinds of CMC differing in average molecular weight (about 1 kDa and 90 kDa) were used to investigate the relationship between molecular weight of conjugated saccharide and saccharide content in the conjugates and degree of functional changes in β-conglycinin. The β-conglycinin-CMC conjugates were purified by dialysis using a dialysis membrane whose molecular weight cutoff is 100 kDa. Composition of the β-conglycinin-low molecular weight (LMW) CMC and β-conglycinin-high molecular weight (HMW) CMC was β-conglycinin: CMC = 1:3.3 and 1:2.1 (weight ratio) respectively which was confirmed by BCA method and phenol sulfuric acid method. Conjugation was confirmed by SDS-PAGE with CBB. Solubility of β-conglycinin in the range of pH4.0-7.0 was much improved by conjugation with both LMW and HMW CMC. Emulsifying property of β-conglycinin at pH5.0 and pH7.0 was much improved by conjugation with HMW CMC and greater improvement was achieved by conjugation with LMW CMC. Immunogenicity of β-conglycinin was decreased by conjugation with LMW CMC.
通过使用水溶性碳二亚胺将β-伴大豆球蛋白与羧甲基纤维素（CMC）偶联，以改善其功能。使用两种平均分子量不同（约1 kDa和90 kDa）的CMC来研究偶联糖的分子量与偶联物中糖含量之间的关系以及β-伴大豆球蛋白功能变化的程度。使用截留分子量为100 kDa的透析膜通过透析法纯化β-伴大豆球蛋白-CMC偶联物。通过BCA法和苯酚硫酸法确定β-伴大豆球蛋白-低分子量（LMW）CMC和β-伴大豆球蛋白-高分子量（HMW）CMC的组成分别为β-伴大豆球蛋白：CMC = 1:3.3和1:2.1（重量比）。通过用考马斯亮蓝染色的SDS-PAGE确认偶联。通过与LMW和HMW CMC偶联，β-伴大豆球蛋白在pH4.0 - 7.0范围内的溶解度得到了很大提高。通过与HMW CMC偶联，β-伴大豆球蛋白在pH5.0和pH7.0时的乳化性能得到了很大改善，而与LMW CMC偶联则实现了更大的改善。通过与LMW CMC偶联，β-伴大豆球蛋白的免疫原性降低。
