A multisubstrate assay for lipases/esterases: Assessing acyl chain length selectivity by reverse-phase high-performance liquid chromatography |
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Authors: | K Divakar Pennathur Gautam |
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Institution: | Centre for Biotechnology, Anna University, Chennai 600 025, India |
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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. |
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Keywords: | Acyl chain length HPLC Lipase Multisubstrate Substrate specificity 4-Nitrophenol |
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