食品科学 ›› 0, Vol. ›› Issue (): 0-0.

• 生物工程 •    下一篇

表面等离子体共振技术研究卵黄抗体对酪氨酸酶与底物相互作用的影响

史佩玉1,隋建新1,曹立民2,林洪1   

  1. 1. 中国海洋大学食品科学与工程学院
    2. 中国海洋大学
  • 收稿日期:2017-01-12 修回日期:2017-11-03 出版日期:2018-01-25 发布日期:2018-01-05
  • 通讯作者: 隋建新 E-mail:suijianxin@ouc.edu.cn

Analysis on the Impact of Specific IgY to the Interaction between Tyrosinase and its Substrate by Surface Plasmon Resonance

Pei-Yu SHI1,Jianxin SUI 1   

  • Received:2017-01-12 Revised:2017-11-03 Online:2018-01-25 Published:2018-01-05
  • Contact: Jianxin SUI E-mail:suijianxin@ouc.edu.cn

摘要: 目的 采用表面等离子共振技术(SPR)测定酪氨酸酶与特异性卵黄抗体(IgY)及其与底物L-多巴的亲和力,探讨特异性IgY对酪氨酸酶与底物亲和力的影响。方法 将裸金芯片进行自组装修饰,以3-巯基丙酸(MPA)作为基底膜对酪氨酸酶进行固定,SPR实时监测特异性IgY及底物L-多巴与酪氨酸酶结合后芯片表面的响应信号变化。将数据导入CLAMP分析软件,进行拟合分析,确定动力学常数。结果与结论 酪氨酸酶与特异性IgY的结和常数明显高于其与底物L-多巴的结和常数,且酪氨酸酶与特异性IgY结合后,后者与底物L-多巴亲和力明显下降,可从其结合动力学角度分析特异性IgY对酪氨酸酶酶活抑制的作用。

关键词: 表面等离子体共振, 酪氨酸酶, 特异性卵黄抗体, L-多巴, 动力学常数

Abstract: Aim The surface plasmon resonance(SPR) technology was used to investigate the impact of Specific immunoglobulin yolk antibody to the interacton between Tyrosinase and its substrate L- Dopa. Methods The bare gold chip was modified by self-assembly and 3-Mercaptopropionic acid (MPA) was used as the basement membrane to immobilize tyrosinase, the SPR response signals of tyrosinse binding to specific IgY and L-dopa were monitored in real time. A CLAMP software was used to process the fitting data to determine the kinetic constants.Results and Conclusions The affinity constant of tyrosinase and specific IgY was higher than that of tyrosinase and L-dopa, and after combined with specific IgY, the constant of tyrosinase and L-dopa decreased significantly, which demonstrated that specific IgY had a remarkable inhibitory effect to tyrosinase activity.

Key words: Surface plasmon resonance(SPR), Tyrosinase, Specific IgY, L- Dopa, Kinetic constants

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