4种酪氨酸酶抑制剂的两两联合作用机制  

Action Mechanisms of Binary Combinations of Four Tyrosinase Inhibitors

作　　者：苏冬雨 刘祎 班晨宇 张瀛心 周素珍 范金波 SU Dongyu;LIU Yi;BAN Chenyu;ZHANG Yingxin;ZHOU Suzhen;FAN Jinbo(College of Food Science and Engineering,Bohai University,Jinzhou 121013,China)

机构地区：[1]渤海大学食品科学与工程学院,辽宁锦州121013

出　　处：《食品科学》2025年第8期8-15,共8页Food Science

基　　金：“十四五”国家重点研发计划重点专项(2023YFF1103502)。

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,结合位点数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的存在并没有改变抑制剂2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,与TYR样品相比,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。差示扫描量热法结果显示,复合抑制剂的热变性熔融温度均低于单抑制剂,表明联合抑制时TYR的热稳定性降低。抑制剂1使抑制剂2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。To investigate the combined inhibitory effects of sophoroside(Sop),puerarin(Pue),mangiferin(Man),and kojic acid(KA)on tyrosinase(TYR),we determined the combination index(CI).Competitive binding experiments were done to evaluate the effect of the order of addition of TYR inhibitors on their binding to TYR,and the mechanism of the combined action of TYR inhibitors was explored by various spectroscopies.The results showed that the combinations of Pue+Sop,Sop+KA,and KA+Man exhibited a synergistic inhibitory effect on TYR,with addition orders of TYR-Pue-Sop,TYRSop-KA,and TYR-KA-Man.All inhibitor combinations statically quenched the fluorescence of TYR,forming a ternary complex.The binding constant of Pue+Sop to TYR was not significantly different from that of Sop,and the number of binding sites was approximately to 1,indicating that the binding sites of the two inhibitors were different.As evidenced by increased binding constant and decreased particle size,Sop and KA promoted the binding of KA and Man to TYR,respectively,which consequently became tighter.The interaction type of one inhibitor was not changed by the presence of another.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 compared with TYR,TYR-Sop-KA and TYR-Pue-Sop tended to transit towards disorder,and the structure of TYR-KA-Man was more unstable.The results of differential scanning calorimetry(DSC)showed that the melting temperature(Tm)of the TYR-inhibitor ternary complexes was lower than that of the binary complexes,indicating decreased thermal stability.In conclusion,an increase in the binding constant of one TYR inhibitor and a decrease in the thermal stability of complexes in the presence of another may be reasons for the synergistic action of the two TYR inhibitors.

关 键 词：联合抑制 槐角苷 葛根素 芒果苷 曲酸 

分 类 号：TS201.2[轻工技术与工程—食品科学]