Colorimetric assay of ethanol using alcohol dehydrogenase from dry bakerʹs yeast

Alcohol dehydrogenases (ADHs) are oxidoreductases present in animal tissues, plants, and microorganisms. These enzymes attract major scientific interest for the evolutionary perspectives, afforded by their wide occurrence in nature, and for their use in synthesis, thanks to their broad substrate specificity and stereoselectivity. In the present study, the standardization of the activity of the alcohol dehydrogenase from bakerʹs yeast was accomplished, and the pH and temperature stability showed, that the enzyme presented a high stability to pH 6.0–7.0 and the thermal stability were completely maintained up to 50 °C during 1 h. The assays of ethanol (detection range 1–5 mM or 4.6 × 10−2 to 23.0 × 10−2 g/L) in different samples in alcoholic beverages, presented a maximum deviation of only 7.2%. The standard curve and the analytic curve of this method meet the conditions of precision, sensitivity, simplicity, and low cost, required for a useable analytical method.