篇名 | Screening Tyrosinase Inhibitors from Traditional Chinese Medicine by Capillary Electrophoresis with Electrophoretically Mediated Microanalysis |
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卷期 | 20:1 |
並列篇名 | 利用電泳中介微分析法篩選中草藥中酪氨酸酶抑製劑 |
作者 | 張文慧 、 呂志華 、 姜廷福 、 王遠紅 、 郭麗萍 |
頁次 | 159-163 |
關鍵字 | 酪氨酸酶 、 篩選 、 中草藥 、 電泳中介微分析 、 electrophorerically mediated microanalysis 、 tyrosinase inhibitor 、 screening 、 traditional Chinese drugs 、 MEDLINE 、 Scopus 、 SCIE |
出刊日期 | 201203 |
本文介紹了電泳中介微分析法篩選酪氨酸酶抑製劑的模型,試驗中優化了酪氨酸酶與底物L-酪氨酸的反應條件,測定了酪氨酸酶的米式常數為0.0532 mM,抑製劑曲酸的抑制機制並計算得到曲酸的抑制常數,與文獻報導相符,證實了該篩選方法的可靠性。並用該方法對20種中草藥萃取物進行了篩選。與傳統的酶抑制劑篩選方法相比,不僅減少了假陽性結果,而且簡化了實驗過程,可用於篩選抑制劑在天然取提物。
Electrophoretically mediated microanalysis (EMMA) is an analytical method using on-column assays. In the setup, substrate and enzyme were individually introduced into the capillary as distinct plugs. The electrophoretic conditions for the enzyme reaction and separation of substrate and product were optimized in this study. The inhibition constant and type of kojic acid (a reference tyrosinase inhibitor) were obtained by this method, which agreed with the literature data. Furthermore, the inhibitory ratios of enzymatic activity (IRE) of 21 traditional Chinese drugs were determined. The EMMA method has superiority over traditional assay methods, in not only minimizing the false-positive results but also simplifying the experimental procedure. Therefore, it could be employed to screen inhibitors from natural sources.