食品科学 ›› 0, Vol. ›› Issue (): 0-0.

• 食品化学 •    下一篇

4-HR与抗坏血酸对中华管鞭虾多酚氧化酶的抑制机理研究

周宇芳1,胡建坤1,郝云彬2,相兴伟1,杨会成1,郑斌1,肖金星3   

  1. 1. 浙江省海洋开发研究院
    2. 舟山市渔业检验检测中心
    3. 浙江大学海洋研究中心
  • 收稿日期:2017-12-06 修回日期:2018-09-19 出版日期:2019-02-25 发布日期:2019-03-05
  • 通讯作者: 相兴伟 E-mail:xxw11086@126.com
  • 基金资助:
    海水甲壳类酚酶抑制剂关键技术研究

Inhibitory Mechanism of 4-Hexylresorcinol and Ascorbic Acid on Polyphenoloxidase from Solenocera Crassicornis

  • Received:2017-12-06 Revised:2018-09-19 Online:2019-02-25 Published:2019-03-05

摘要: 研究无硫复合抑制剂主要作用成分4-己基间苯二酚(4-hexylresorcinol,4-HR)、抗坏血酸对中华管鞭虾多酚氧化酶(polyphenoloxidase,PPO)活性的抑制作用,并从抑制剂化学结构及动力学模型出发分析推断其抑制机理。结果表明,4-HR、抗坏血酸均能抑制PPO酶活性,且对酶促反应有延滞作用,两者半抑制浓度分别为0.072、2.02 mmol/L,对中华管鞭虾PPO酶的抑制作用都表现为竞争性抑制;化学动力学模拟分析显示4-HR、抗坏血酸均可与PPO酶双铜活性中心结合,其中4-HR可生成一种高稳定的多酚氧化酶络合物,持续有效地消耗PPO酶。结论,4-HR、抗坏血酸均能有效抑制中华管鞭虾PPO酶酶促反应,其中4-HR抑制效果更显著。本试验为无硫复合抑制剂在海水虾捕捞与加工过程中的应用推广提供了理论依据。

关键词: 中华管鞭虾, 多酚氧化酶(PPO), 4-己基间苯二酚(4-HR), 抗坏血酸, 化学动力学模拟

Abstract: A sulfur-free compound preservative, the main effective constituents of which were 4-hexylresorcinol (4-HR) and ascorbic acid (AA), was developed in view of seawater shrimp melanosis, sulfite abuse and short shelf-life. The purpose of this study was to investigate the inhibitory effects of 4-hexylresorcinol (4-HR) and AA on polyphenoloxidase (PPO) purified from Solenocera Crassicornis. In addtion, the analysis of chemical structural model, dynamic simulation, and interaction energy analysis of the inhibitors were employed to determine the inhibitory mechanism. The results showed that 4-HR and AA could inhibit the enzyme activity of PPO significantly and provoke lag periods of enzymatic reaction. With the increase of 4-HR and AA concentrations, the residual enzyme activity of PPO decreased. The median inhibitory concentrations (IC50) were erstimated to be 0.072 mmol/L for 4-HR and 2.02 mmol/L for AA, respectively. 4-HR and AA both exhibited competitive inhibitory pattern on PPO. According to the results of Chemical kinetics simulation analysis, 4-HR and AA were combined with di-copper binding centre of PPO. And 4-HR would consume PPO effectively as forming a high stable complex. In conclusion, 4-HR and AA have been shown to be effective in the control of enzymatic reaction rate, and 4-HR was more effective. The results could provide some theoretical support for the application of sulfur-free compound preservative on the fishing and processing of seawater shrimp.

Key words: Solenocera Crassicornis, polyphenoloxidase (PPO), 4-hexylresorcinol (4-HR), ascorbic acid, chemical kinetics simulation analysis

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