Jump to main content
Jump to site search
Access to RSC content Close the message box

Continue to access RSC content when you are not at your institution. Follow our step-by-step guide.


Issue 4, 2011
Previous Article Next Article

Optimization of a colorimetric assay for yeast lipase activity in complex systems

Author affiliations

Abstract

The present work describes a simple and sensitive spectrophotometric method based on the release of p-nitrophenyl butyrate (p-NPB) for the estimation of lipase activity. The method was optimized and validated in biphasic complex media containing methyl ricinoleate (MR) or olive mill wastewater (OMW), although it may be used for other biphasic media. Reaction mixtures containing substrate (2.63 mM p-NPB in sodium acetate buffer, 0.05 M and pH 5.6, with 4% (v/v) Triton X-100) and lipase are incubated at 37 °C during 15 minutes. After this time, 2 mL of acetone are added to stop the reaction and the corresponding absorbances are measured at 405 nm in a microplate reader. A linear response was obtained for Candida rugosalipase concentrations in the range of 0.0054 to 0.1 g L−1 and 0.093 to 0.5 g L−1 for MR and OMW media, respectively. The method revealed to be more sensitive for MR medium. This conclusion was corroborated by the detection and quantification limits, which were smaller for MR than for OMW medium. The method is precise for both tested media, according to the Horwitz criterion. Besides the simplicity of this method, it is sensitive and precise, and could be adopted for routine analysis, as a tool for screening and detection of lipase activity. In addition, the fact of measuring the absorbances of the samples in a microplate reader, allows to analyze a great number of samples at the same time and to achieve a considerable time saving.

Graphical abstract: Optimization of a colorimetric assay for yeast lipase activity in complex systems

Back to tab navigation

Article information


Submitted
08 Nov 2010
Accepted
03 Feb 2011
First published
24 Feb 2011

Anal. Methods, 2011,3, 1008-1013
Article type
Technical Note

Optimization of a colorimetric assay for yeast lipase activity in complex systems

N. Gomes, C. Gonçalves, M. García-Román, J. A. Teixeira and I. Belo, Anal. Methods, 2011, 3, 1008
DOI: 10.1039/C0AY00680G

Social activity

Search articles by author

Spotlight

Advertisements