Development of a high-throughput liquid state assay for lipase activity using natural substrates and rhodamine B

A fluorescence-based assay for the determination of lipase activity using rhodamine B as an indicator, and natural substrates such as olive oil, is described. It is based on the use of a rhodamine B–natural substrate emulsion in liquid state, which is advantageous over agar plate assays. This high-t...

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Bibliographic Details
Published in:Analytical biochemistry 2016-03, Vol.496, p.25-29
Main Authors: Zottig, Ximena, Meddeb-Mouelhi, Fatma, Beauregard, Marc
Format: Article
Language:English
Subjects:
Colorimetry
Fluorescence
High-Throughput Screening Assays
Lipase - metabolism
Lipase activity
Olive oil
Rhodamine B
Rhodamines - metabolism
Substrate Specificity
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Summary:A fluorescence-based assay for the determination of lipase activity using rhodamine B as an indicator, and natural substrates such as olive oil, is described. It is based on the use of a rhodamine B–natural substrate emulsion in liquid state, which is advantageous over agar plate assays. This high-throughput method is simple and rapid and can be automated, making it suitable for screening and metagenomics application. Reaction conditions such as pH and temperature can be varied and controlled. Using triolein or olive oil as a natural substrate allows monitoring of lipase activity in reaction conditions that are closer to those used in industrial settings. The described method is sensitive over a wide range of product concentrations and offers good reproducibility.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2015.11.020