title: A multisubstrate assay for lipases/esterases: assessing acyl chain length selectivity by reverse-phase high-performance liquid chromatography.
abstract: 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.
authors: Centre for Biotechnology, Anna University, Chennai 600 025, India.
authors: K
authors: K
authors: Divakar
authors: Centre for Biotechnology, Anna University, Chennai 600 025, India. Electronic address: pgautam@annauniv.edu.
authors: Pennathur
authors: P
authors: Gautam
mesh_terms: D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D057075:Enzyme Assays / Q000379:methods*; D004950:Esterases / Q000378:metabolism*; D004952:Esters; D005227:Fatty Acids / Q000737:chemistry / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D008049:Lipase / Q000378:metabolism*; D009596:Nitrophenols / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity
chemical_list: D004952:Esters; D005227:Fatty Acids; D009596:Nitrophenols; C024719:4-nitrophenyl; D004950:Esterases; D008049:Lipase
keywords: 4-Nitrophenol; Acyl chain length; HPLC; Lipase; Multisubstrate; Substrate specificity