Use of alcohol oxidase to measure the methanol produced during the hydrolysis of D- and L-methyl-3-hydroxybutyric acid.
An enzymatic assay for the measurement of methanol has been developed. The assay uses alcohol oxidase and peroxidase coupled to the oxidation of 2,2'-azino-di-(3-ethyl)-benzthiazoline-6-sulfonic acid as the chromogen. The assay is linear up to 50 nmol of methanol in a 200-microliters sample and sensitive; 1.25 nmol of methanol in a 200-microliters sample can be measured. The assay is rapid and measurements can be made at any convenient time between 15 min and 4 h after initiation of the reaction. The assay shows highest activity with methanol but significant activity with other primary alcohols up to 1-butanol. Little activity is shown with secondary alcohols and diols. We have used this assay to follow the hydrolysis of the two isomers of the methyl ester of 3-hydroxybutyric acid.
D000429:Alcohol Oxidoreductases; D006868:Hydrolysis; D006885:Hydroxybutyrates; D000432:Methanol / Q000032:analysis*; D010544:Peroxidases; D013053:Spectrophotometry
D006885:Hydroxybutyrates; C016579:2-methyl-3-hydroxybutyric acid; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D010544:Peroxidases; D000432:Methanol