四种酪氨酸酶抑制剂的两两联合作用机制

摘要/Abstract

摘要： 为探究槐角苷(sophoricoside，Sop)、葛根素(puerarin，Pue)、芒果苷(mangiferin，Man)、曲酸(kojic acid，KA)对酪氨酸酶(tyrosinase，TYR)的联合抑制作用，通过联合作用指数、竞争结合实验确定协同抑制作用组合及其添加顺序，借助多光谱法探究联合作用机制。结果表明：Pue+Sop、Sop+KA、KA+Man组合发挥协同抑制作用，添加顺序为TYR-Pue-Sop、TYR-Sop-KA、TYR-KA-Man。联合作用时均发生静态猝灭，即形成了酶-双抑制剂三元复合物；TYR-Pue-Sop较TYR-Sop的结合常数无较大差别，n≈1，说明两者结合位点不同；结合常数的增加和粒径的降低表明Sop、KA分别促进了TYR-KA、TYR-Man的结合，使其结合更加紧密。抑制剂1(Inhibitor 1，I1)的存在并没有改变抑制剂2(Inhibitor 2，I2)原有的作用力类型，TYR-Pue-Sop以静电相互作用为主，TYR-Sop-KA以氢键和范德华力为主，TYR-KA-Man以疏水相互作用为主要驱动力。红外光谱显示，TYR-Sop-KA、TYR-Pue-Sop趋于向无序化方向转变，TYR-KA-Man的结构更加不稳定。差示扫描量热法(differential scanning calorimetry，DSC)结果显示，复合抑制剂的Tm值均低于单抑制剂，表明联合抑制时TYR的热稳定性降低。结论：I1使I2结合常数的增加及热稳定性的降低是双抑制剂产生协同作用的一些原因。

关键词: 联合抑制, 槐角苷, 葛根素, 芒果苷, 曲酸

Abstract: To investigate the combined inhibitory effects of sophoroside (Sop), puerarin (Pue), mangiferin (Man), and kojic acid (KA) on tyrosinase (TYR), the combination and adding sequence of synergistic inhibition were determined by combined effect index and competitive binding experiment, and the combined action mechanism were explored by multi-spectroscopy. The results showed that the combination of Pue+Sop, Sop+KA, and KA+Man exhibited a synergistic inhibitory effect, with the addition order being TYR-Pue-Sop, TYR-Sop-KA, and TYR-KA-Man. Static quenching occurred during the combined action, that was, the enzyme-double inhibitor ternary complex were formed. The binding constant of TYR-Pue-Sop was not significantly different from that of TYR-Sop, with n ≈ 1, indicating that the binding sites of the two are different. The increase in binding constant and the decrease in particle size indicated that Sop and KA respectively promoted the binding of TYR-KA and TYR-Man, making their binding more compact. The existence of inhibitor 1 (I1) did not change the original force type of inhibitor 2 (I2). Electrostatic interaction was dominant in TYR-Pue-Sop, hydrogen bond and van der Waals force in TYR-Sop-KA and hydrophobic interaction in TYR-KA-Man. Infrared spectroscopy showed that TYR-Sop-KA and TYR-Pue-Sop tended to transition towards disorder, and the structure of TYR-KA-Man was more unstable. The results of differential scanning calorimetry (DSC) showed that the Tm values of the composite inhibitor were lower than those of the single inhibitor, indicating that the thermal stability of TYR decreased when combined inhibition. Conclusion: the increase in the binding constant and the decrease of thermal stability of I2 caused by I1 are some reasons for the synergistic effect of double inhibitors.

Key words: combined inhibition, sophoricoside(Sop), puerarin(Pue), mangiferin(Man), kojic acid(KA)

中图分类号:

TS201.4

苏冬雨刘祎班晨宇张瀛心周素珍范金波. 四种酪氨酸酶抑制剂的两两联合作用机制[J]. 食品科学.

Jin-Bo FAN. the Pairwise Combined Mechanism of Four Tyrosinase Inhibitors[J]. FOOD SCIENCE.

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