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

作者：Divakar K; Gautam Pennathur^*

来源：Analytical Biochemistry, 2014, 448: 38-40.

DOI：10.1016/j.ab.2013.11.031

• Acyl chain length
• HPLC
• Lipase
• Multisubstrate
• Substrate specificity
• 4-Nitrophenol

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.

• 出版日期2014-3-1