模拟酶荧光素抑制剂的筛选及分析应用研究

Nature enzyme, such as HRP, have been exploited for widespread applications, owing to their high efficiency and specificity under mild conditions. Considering the intrinsic drawbacks of natural enzymes, such as high cost in large scale production, poor stability of catalytic activities against denaturation, digestion or the variations of environmental conditions, researchers have therefore endeavoured to develop artificial enzymes as viable alternatives. Construction of nanozymes has been focus of research over past decade. Compared with natural enzymes, these nanozymes are more stable against denaturation or protease digestion, low in cost, and easy to prepare, store and treat. Because most of them are spontaneous aggregation in aqueous solutions or under high salt conditions to lose the catalytical activity. Moreover, the preparation of water-soluble nanozymes and the loading steps of nanomaterials are complicated and time-consuming. The low stability under high salt conditions and unsatisfied batch-to-batch consistence of peroxidase-like activity has unfortunately restricted their further applications in a way. Therefore, it is highly desirable to search for a new mimetic enzyme that possesses highly repeatable activity and highly stable against harsh condition with low cost. We found fluorescein possesses mimic peroxidase and visible-light-induced oxidase mimic property. As a new class of enzyme mimics, inhibitors of small molecule-based artificial enzymes have not been investigated. Due to a quite different composition and structure from nature enzymes and nanozymes that they have distinct inhibitors. Firstly, screening of inhibitors of artificial enzyme of fluorescein and inhibition mechanisms will be studied. Secondly, we will investigate evaluation of antioxiant behavior. Thirdly, new colorimetric strategies based on the analyte tuned peroxidase/oxidase-mimicking activity of fluorescein alternatives will be developed. This work not only will offer a new insight into screening of inhibitors of enzyme mimetics, but also broaden the scope of analytical application.

天然酶，如辣根过氧化物酶（HRP），由于其高的催化效率和特异性而被广泛应用在各个领域。但其也存在生产成本高、易失活和变性等一些固有缺陷，因此研究者一直致力于开发能克服上述缺陷的模拟酶。在过去的十年里，纳米酶的构建已经成为研究热点。但它们大多数在水溶液中或高盐条件下长时间放置易于团聚而失去催化活性。另外，大部分的纳米酶很难大批量的制备，而且同种纳米酶的催化活性也存在批次间的差异。我们发现，有机小分子物质——荧光素具有过氧化物模拟酶和可见光诱导的氧化物模拟酶活性，提出了发展小分子模拟酶的思路。作为小分子，荧光素活性不易受环境影响、无批间差异和可通过改变基团调控。荧光素作为新型的模拟酶，由于与天然酶或其它人工酶的组成和结构不同，所以其抑制剂也会有所不同。基于此，我们拟完成：筛选荧光素模拟酶抑制剂并探讨抑制机理；研究荧光素模拟酶对抗氧化剂的抗氧化能力；设计荧光素基模拟酶探针构建新的分析方法。

天然酶，如辣根过氧化物酶（HRP），由于其高的催化效率和特异性而被广泛应用在各个领域。但其也存在生产成本高、易失活和变性等一些固有缺陷，因此研究者一直致力于开发能克服上述缺陷的模拟酶。在过去的十年里，纳米酶的构建已经成为研究热点。但它们大多数在水溶液中或高盐条件下长时间放置易于团聚而失去催化活性。另外，大部分的纳米酶很难大批量的制备，而且同种纳米酶的催化活性也存在批次间的差异。我们发现，有机小分子物质——荧光素具有过氧化物模拟酶和可见光诱导的氧化物模拟酶活性，提出了发展小分子模拟酶的思路。作为小分子，荧光素活性不易受环境影响、无批间差异和可通过改变基团调控。荧光素作为新型的模拟酶，由于与天然酶或其它人工酶的组成和结构不同，所以其抑制剂也会有所不同。基于此，我们完成了：筛选荧光素模拟酶抑制剂并探讨抑制机理；研究荧光素模拟酶对抗氧化剂的抗氧化能力；设计荧光素基模拟酶探针构建新的分析方法。具体内容如下：.1、5-硝基荧光素过氧化物模拟酶活性的探索.2、基于5-硝基荧光素的过氧化物模拟酶活性的光度法检测头孢呋辛钠.3、头孢呋辛钠对5-硝基荧光素过氧化物模拟酶活性的抑制机理和动力学.4、不同基团对卤代荧光素在可见光诱导下模拟氧化酶活性的影响.5、不同官能团对荧光素衍生物光催化性能的影响.6、6-硝基荧光素过氧化物模拟酶活性的发现.7、二碘荧光素过氧化物模拟酶的性能研究.8、基于荧光素及其衍生物的过氧化物模拟酶活性用于生物检测和生物抗氧化行为的评估.9、羧基荧光素模拟过氧化物酶比色法快速检测水果和维生素C片中抗坏血酸.10、基于荧光素衍生物过氧化物模拟酶的活性比色检测L-半胱氨酸.11、基于优良的5(6)-氨基荧光素类过氧化物模拟酶性能灵敏检测酒石酸.12、比色法检测虎杖痛酊中白藜芦醇.13、没食子酸抑制荧光素衍生物的过氧化物模拟酶活性的比色分析.14、基于5(6)-羧基荧光素过氧化物模拟酶活性的硫离子检测.15、基于羧基荧光素模拟酶活性比色法检测茶多酚.16、氨基荧光素作为过氧化物模拟酶用于谷胱甘肽的检测