Enzyme assay and activity fingerprinting of hydrolases with the red-chromogenic adrenaline test.
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.
Fluxá
Viviana S
VS
Department of Chemistry and Biochemistry, University of Berne, Freiestrasse 3, Berne CH-3012, Switzerland.
Wahler
Denis
D
Reymond
Jean-Louis
JL
D000323:Adrenochrome / Q000032:analysis* / Q000737:chemistry; D001234:Aspergillus niger / Q000201:enzymology; D003124:Colorimetry / Q000379:methods*; D004837:Epinephrine / Q000737:chemistry; D004851:Epoxide Hydrolases / Q000737:chemistry / Q000378:metabolism*; D004950:Esterases / Q000737:chemistry / Q000378:metabolism*; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*
D000323:Adrenochrome; D004950:Esterases; D008049:Lipase; D004851:Epoxide Hydrolases; D004837:Epinephrine