针对导致黑色素沉淀的酪氨酸酶,制备了特异性卵黄抗体Fab'片段,研究其对酶活力的抑制效果并从动力学角度进行分析.结果表明,胃蛋白酶法制备Fab'片段的最适条件为:pH 4.0、反应比(卵黄抗体-胃蛋白酶)1∶150 (mg/U)、反应时间8h.经免疫亲和柱纯化后可得理化性质稳定、纯度大于95％的Fab'片段;该片段通过非竞争性结合抑制酪氨酸酶活力,IC50为11.16 μmol/L,结合常数为5.79×105,抑制效果相比完整的卵黄抗体分子大大提高.本研究可为卵黄抗体Fab'片段作为潜在的酶活力抑制剂在食品、化妆品和医药等行业中应用提供理论参考.%The production of Fab'fragment from specific egg yolk antibody against tyrosinase and its inhibitory activity on tyrosinase were investigated in this paper.The optimal conditions were obtained as follows:pH 4.0,ratio of egg yolk antibody to pepsin 1∶150 (mg/U),and reaction time 8 h.Fab'fragment purified by immunoaffmity column chromatography to a purity of over 95％ exhibited good physicochemical stability.It was demonstrated as a non-competitive inhibitor with an IC50value of 11.16 tmol/L and a binding constant of 5.79 × 105,whose inhibitory effect was much higher than that of IgY.The results of this study can provide a theoretical basis for the application of egg yolk antibody Fab'fragments in food,cosmetics and pharmaceutical industries as a potential enzyme inhibitor.
展开▼
