Enzyme assay and activity fingerprinting of hydrolases with the red-chromogenic adrenaline test

Nature Protocols
Viviana S FluxáJean-Louis Reymond

Abstract

The adrenaline test for enzymes is a colorimetric enzyme assay based on the quantification of periodate-sensitive reaction products such as 1,2-diols and 1,2-aminoalcohols by back-titration of the oxidant with adrenaline to produce adrenochrome as an easily detectable red product. The test uses commercial reagents and is suitable for screening the activity of various hydrolases. It is demonstrated here for testing epoxide hydrolases, lipases and esterases, and for activity fingerprinting of these enzymes across substrate series. The complete assay requires 2-3 h.

References

Aug 31, 2002·Chembiochem : a European Journal of Chemical Biology·Jean-Louis Reymond, Denis Wahler
Dec 12, 2002·Journal of the American Chemical Society·Cleo M SalisburyJonathan A Ellman
Dec 20, 2002·Angewandte Chemie·Denis Wahler, Jean-Louis Reymond
Jul 10, 2003·Proceedings of the National Academy of Sciences of the United States of America·Dhaval N Gosalia, Scott L Diamond
Sep 10, 2004·Journal of the American Chemical Society·Jean-Philippe Goddard, Jean-Louis Reymond
Jan 15, 2005·Analytical Chemistry·Peter Babiak, Jean-Louis Reymond
Apr 6, 2005·Bioorganic & Medicinal Chemistry Letters·Mahesh UttamchandaniShao Q Yao
Oct 27, 2005·Analytical Chemistry·James P Smith, Vicki Hinson-Smith
Aug 2, 2006·Molecular BioSystems·Yang Yongzheng, Jean-Louis Reymond
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Jan 12, 2007·Molecular BioSystems·Johann Grognux, Jean-Louis Reymond
Jan 30, 2007·Organic Letters·Sungjin Park, Injae Shin

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Citations

Jul 11, 2009·Applied Microbiology and Biotechnology·Won Jae Choi
Nov 23, 2010·Biotechnology Letters·Abeer Ahmed QaedZhong-Liu Wu
Mar 24, 2012·Chemical Communications : Chem Comm·Cristian MacoveiFrédéric Taran
Dec 17, 2008·Chemical Communications : Chem Comm·Jean-Louis ReymondNoélie Maillard
Sep 3, 2020·Acta Crystallographica. Section D, Structural Biology·Gabriela D Tormet-GonzálezMarcio Vinicius Bertacine Dias
Jul 29, 2011·Organic & Biomolecular Chemistry·Suttiporn PinijsuwanKurt V Gothelf

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