摘要
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8 h。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,IC_(50)为11.16μmol/L,结合常数为5.79×10~5,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 immunoaffinity column chromatography to a purity of over 95% exhibited good physicochemical stability.It was demonstrated as a non-competitive inhibitor with an IC（50） value of 11.16 μmol/L and a binding constant of 5.79 × 10~5,whose inhibitory effect was much higher than that of Ig Y.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.
作者
钟欣欣
曹立民
林洪
史佩玉
隋建新
ZHONG Xinxin;CAO Limin;LIN Hong;SHI Peiyu;SUI Jianxin(College of Food Science and Engineering, Ocean University of China, Qingdao 266003, Chin)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2018年第2期119-123,共5页
Food Science
基金
中央高校基本科研业务费专项(201413061)
山东省农业应用创新专项
关键词
酪氨酸酶
卵黄抗体
蛋白酶解
Fab’片段
活性抑制
tyrosinase
egg yolk antibody
enzymatic proteolysis
fragment antigen binding （Fab’ fragment）
enzyme activity inhibition
