Yoshida Tadashi, Hamaji Ikumi, Hashimoto Takeshi, Matsumoto Takuya, Hattori Makoto
Department of Applied Biological Science, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-Cho, Fuchu, Tokyo 183-8509 Japan.
Cytotechnology. 2023 Apr;75(2):143-151. doi: 10.1007/s10616-022-00568-6. Epub 2023 Jan 5.
β-Conglycinin was conjugated with ε-polylysine (PL) by means of microbial transglutaminase (MTGase) to improve its function. The β-conglycinin-PL conjugate was purified by dialysis. Composition of the β-conglycinin-PL was β-conglycinin:PL = 1:18 (molar ratio) which was confirmed by amino acid analysis. The β-conglycinin-PL was further conjugated with dextran (Dex) by the Maillard reaction. The β-conglycinin-PL-Dex conjugate was purified by dialysis. Conjugation was confirmed by SDS-PAGE and PAS staining. Composition of the β-conglycinin-PL-Dex was β-conglycinin-PL:Dex = 1:41 (molar ratio) which was confirmed by UV spectra measurement and phenol sulfuric acid method. Solubility of β-conglycinin in the acidic range was much improved by conjugation with PL and further improved by further conjugation with Dex. Emulsifying property of β-conglycinin in acidic pH range was much improved by conjugation with PL and Dex. Immunogenicity of β-conglycinin was decreased by conjugation with PL and Dex.
通过微生物转谷氨酰胺酶（MTGase）将β-伴大豆球蛋白与ε-聚赖氨酸（PL）偶联以改善其功能。通过透析纯化β-伴大豆球蛋白-PL偶联物。通过氨基酸分析证实β-伴大豆球蛋白-PL的组成是β-伴大豆球蛋白：PL = 1:18（摩尔比）。通过美拉德反应将β-伴大豆球蛋白-PL与葡聚糖（Dex）进一步偶联。通过透析纯化β-伴大豆球蛋白-PL-Dex偶联物。通过SDS-PAGE和PAS染色确认偶联。通过紫外光谱测量和苯酚硫酸法证实β-伴大豆球蛋白-PL-Dex的组成是β-伴大豆球蛋白-PL：Dex = 1:41（摩尔比）。通过与PL偶联，β-伴大豆球蛋白在酸性范围内的溶解度大大提高，通过进一步与Dex偶联进一步提高。通过与PL和Dex偶联，β-伴大豆球蛋白在酸性pH范围内的乳化性能大大提高。通过与PL和Dex偶联，β-伴大豆球蛋白的免疫原性降低。
