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Updated: Sep 25, 2026

Defining Substrate Specificities for Lipase and Phospholipase Candidates
Published on: November 23, 2016
Lipase Activity, Analytical Methods: Brief Review and Gold Standards
Josu López-Fernández1, Arnau Gasset1, Marina Guillén1
1Department of Chemical, Biological and Environmental Engineering, School of Engineering, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain.
Abstract:
Amid the growing demand for sustainable practices in industry, lipases emerge as crucial players, constituting a significant share of the enzyme market. Consequently, the measurement of lipase activity remains essential for their effective utilization. In this chapter, quantitative techniques are reviewed, with a focus on hydrolytic methods. Titrimetric approaches, relying on free fatty acid titration, and spectrophotometric methods using soluble p-nitrophenyl esters are highlighted as gold standards. Challenges in lipase activity measurement, including substrate specificity and assay conditions, are discussed. Detailed protocols for lipase activity assessment using oil and p-nitrophenyl butyrate as substrates-visible spectrophotometry and titrimetry-are provided. Ultimately, this chapter underscores the importance of robust methodologies in unlocking the full potential of lipases for diverse industrial applications.

