Divakar K, Gautam Pennathur
Centre for Biotechnology, Anna University, Chennai 600 025, India.
Centre for Biotechnology, Anna University, Chennai 600 025, India.
Anal Biochem. 2014 Mar 1;448:38-40. doi: 10.1016/j.ab.2013.11.031. Epub 2013 Dec 6.
Lipases and esterases are hydrolytic enzymes and are known to hydrolyze esters with unique substrate specificity and acyl chain length selectivity. We have developed a simple competitive multiple substrate assay for determination of acyl chain length selectivity of lipases/esterases using RP-HPLC with UV detection. A method for separation and quantification of 4-nitrophenyl fatty acid esters (C4-C18) was developed and validated. The chain length selectivity of five lipases and two esterases was determined in a multisubstrate reaction system containing equimolar concentrations of 4-nitrophenyl esters (C4-C18). This assay is simple, reproducible, and a useful tool for determining chain length selectivity of lipases/esterases.
脂肪酶和酯酶是水解酶，已知它们能以独特的底物特异性和酰基链长度选择性水解酯类。我们开发了一种简单的竞争性多底物测定法，用于使用带紫外检测的反相高效液相色谱法（RP-HPLC）测定脂肪酶/酯酶的酰基链长度选择性。建立并验证了一种分离和定量4-硝基苯基脂肪酸酯（C4-C18）的方法。在含有等摩尔浓度4-硝基苯基酯（C4-C18）的多底物反应体系中测定了五种脂肪酶和两种酯酶的链长度选择性。该测定法简单、可重复，是测定脂肪酶/酯酶链长度选择性的有用工具。
