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通过制备羧甲基纤维素生物共轭物改善β-伴大豆球蛋白的功能。

Functional improvements in β-conglycinin by preparing bioconjugates with carboxymethyl cellulose.

作者信息

Hataishi Yui, Tanaka Aya, Ishizuka Misaki, Mizobuchi Hibine, Yoshida Tadashi, Hattori Makoto

机构信息

Department of Applied Biological Science, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-Cho, Fuchu City, Tokyo 183-8509 Japan.

出版信息

Cytotechnology. 2025 Feb;77(1):6. doi: 10.1007/s10616-024-00664-9. Epub 2024 Nov 21.

Abstract

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.

摘要

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。

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