Abstract
“Mureka”(an unpleasant smell developed in nonpasteurized sake) is composed of i-valeraldehyde (i-Val), ethyl i-valerate and 1, 1-diethoxy-3-methylbutane. i-Val is the main component of “Mureka” and formed by an oxidative enzyme reaction on i-Amyl alcohol (i-AmOH). The method for measuring the activity of the “Mureka”-forming enzyme using chromogenic hydrogen donors and peroxidase (POD) was studied. Among various chromogenic hydrogen donors tested, Leuo Patent Blue Violet (LPBV) was most sensitive and stable for measuring the enzyme activity. The reaction mixture contained 0.3ml of the enzyme solution and 2.2ml of substrate solution containing 50mM PIPES buffer, pH 6.5, 0.2mM LPBV, 22 unit POD from horseradish and 1000ppm i-AmOH. The reaction mixture was incubated at 35° for 3hr, and the increase in absorvance at 639nm was measured. It was suggested that the method could also be applied to the measurement of the activity of alcohol oxidase.
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