Chapter title |
Functional-Based Screening Methods for Detecting Esterase and Lipase Activity Against Multiple Substrates
|
---|---|
Chapter number | 4 |
Book title |
Lipases and Phospholipases
|
Published in |
Methods in molecular biology, August 2018
|
DOI | 10.1007/978-1-4939-8672-9_4 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8671-2, 978-1-4939-8672-9
|
Authors |
Dolores Reyes-Duarte, Cristina Coscolín, Mónica Martínez-Martínez, Manuel Ferrer, Humberto García-Arellano, Reyes-Duarte, Dolores, Coscolín, Cristina, Martínez-Martínez, Mónica, Ferrer, Manuel, García-Arellano, Humberto |
Abstract |
Functional screens have been extensively used for searching native enzymes or mutant variants in clone libraries. Esterases and lipases are the most retrieved enzymes, because they are within the more demanded industrial enzymes and because a number of simple and generic screening methods can be applied for their screen. Here, we describe the use of a generic pH indicator assay protocol which unambiguously allows detecting in high-throughput manner esterase and lipase activity and quantifying specific activities using an ester concentration above 0.5 mM. The described method is simple and generic to allow the selection of esterases and lipases targeting desired esters. |
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