A multisubstrate assay for lipases/esterases: Assessing acyl chain length selectivity by reverse-phase high-performance liquid chromatography

被引：4

作者：

Divakar, K. ^{[1
]}

Gautam, Pennathur ^{[1
]}

机构：

[1] Anna Univ, Ctr Biotechnol, Madras 600025, Tamil Nadu, India

来源：

ANALYTICAL BIOCHEMISTRY | 2014年 / 448卷

关键词：

Acyl chain length; HPLC; Lipase; Multisubstrate; Substrate specificity; 4-Nitrophenol; NITROPHENYL PALMITATE ASSAY; ORGANIC MEDIA; LIPASE ACTIVITY; FATTY-ACIDS; ESTERIFICATION; SPECIFICITY; SOLVENT;

D O I：

10.1016/j.ab.2013.11.031

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 (C-4-C-18) 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 (C-4-C-18). This assay is simple, reproducible, and a useful tool for determining chain length selectivity of lipases/esterases. (C) 2013 Elsevier Inc. All rights reserved.

引用

页码：38 / 40

页数：3

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