Yang Yongzheng, Babiak Peter, Reymond Jean-Louis
Department of Chemistry & Biochemistry, University of Berne, Switzerland.
Org Biomol Chem. 2006 May 7;4(9):1746-54. doi: 10.1039/b601151a. Epub 2006 Apr 3.
FRET-based fluorogenic substrates for lipases and esterases were prepared in four steps from commercially available building blocks. The substrates are pyrenebutyric acid monoesters of aliphatic 1,2-diols bearing a dinitrophenylamino group as a quencher. The most enzyme-reactive substrate is ester 2a. The substrates do not show any measurable background reaction in the absence of enzyme even at pH 11, but react fast and specifically with lipases and esterases. These substrates offer an unprecedented and practical solution to the long-standing problem of a simple yet efficient high-throughput screening tool for lipase activities under basic conditions.
基于荧光共振能量转移（FRET）的脂肪酶和酯酶荧光底物由市售的起始原料分四步制备而成。这些底物是带有二硝基苯基氨基作为猝灭剂的脂肪族1,2 -二醇的芘丁酸单酯。酶反应活性最高的底物是酯2a。即使在pH 11的条件下，这些底物在没有酶的情况下也不会显示出任何可测量的背景反应，但能与脂肪酶和酯酶快速且特异性地反应。这些底物为长期存在的在碱性条件下简单而高效的脂肪酶活性高通量筛选工具这一问题提供了前所未有的实用解决方案。
