合成染料与染料脱色过氧化物酶Il-DyP4相互作用的光谱分析  

Exploring the interaction between synthetic dyes and dye decolorization peroxidase of Il-DyP4 by spectral analysis

作　　者：严凤燕 杨阳[1] 荚荣[1] YAN Fengyan;YANG Yang;JIA Rong(School of Life Sciences,Anhui University,Hefei230601,China)

机构地区：[1]安徽大学生命科学学院,安徽合肥230601

出　　处：《安徽大学学报（自然科学版）》2024年第3期91-101,共11页Journal of Anhui University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(31970100)。

摘　　要：利用紫外-可见吸收光谱法和荧光光谱法,分析蒽醌类染料(reactive blue 19(RB19),reactive blue 4(RB4))、偶氮类染料(reactive violet 5(RV5),reactive black 5(RB5))及底物2,2′-联氮-双(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)与染料脱色过氧化物酶Il-DyP4之间的相互作用.紫外-可见吸收光谱法结果表明,Il-DyP4和上述底物之间均形成了复合物;荧光光谱法结果表明,上述底物均使Il-DyP4产生了荧光淬灭效应,且淬灭类型为静态淬灭,符合紫外-可见吸收光谱的结果.根据双对数方程■,计算出RB19,RB4,RV5,RB5,ABTS与Il-DyP4相互作用的表观结合常数(Ka)分别为:9.386 2×10^(4)L·mol^(-1)(298 K),5.615 7×10^(6) L·mol^(-1)(298 K),1.926 2×10^(6) L·mol^(-1)(288 K),5.277 8×10^(5) L·mol^(-1)(298 K),2.234 6×10^(5) L·mol^(-1)(298 K).热力学参数分析结果表明,上述底物与Il-DyP4的结合依赖于疏水相互作用,是自发的吸热过程.In this work,the authors investigated the interaction between dye-decolorizing peroxidase(Il-DyP4)and dyed substrates using ultraviolet-visible(UV-Vis)absorption spectrum and fluorescence spectrum,which included anthraquinone dyes(reactive blue 19,RB19;reactive blue 4,RB4),azo dyes(reactive purple 5,RV5;reactive black 5,RB5),and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt(ABTS).The UV-Vis absorption spectrum showed that the substrates studied in this work could form complexes with Il-DyP4.Furthermore,the fluorescence spectrum also showed that the studied dyed substrates had strong fluorescence quenching effect on Il-DyP4,and the type of quenching was static quenching which depended on the complex formation.The apparent binding constants(K_a)of RB19,RB4,RV5,RB5,ABTS interacting with Il-DyP4 were calculated at different temperatures based on the double logarithmic equation as follows:9.3862×10^(4)L·mol^(-1)(298 K),5.6157×10^(6) L·mol^(-1)(298 K),1.9262×10^(6) L·mol^(-1)(288 K),5.2778×10^(5) L·mol^(-1)(298 K),2.2346×10^(5) L·mol^(-1)(298 K).According to the analysis of thermodynamic parameters,the interaction between Il-DyP4 and the substrate depended on hydrophobic interaction,which was a spontaneous endothermic process.

关 键 词：染料脱色过氧化物酶 紫外-可见吸收光谱 荧光光谱 染料类底物 疏水相互作用 

分 类 号：Q814.9[生物学—生物工程]