Abstract
Peroxygenase is a plant enzyme that catalyzes the oxidation of a double bond to an epoxide in a stereospecific and enantiofacially selective manner. A microsomal fraction containing peroxygenase was prepared from oat (Avena sativa) seeds and the enzyme immobilized onto a hydrophobic membrane. The enzymatic activity of the immobilized preparation was assayed in 1 h by measuring epoxidation of sodium oleate (5 mg) in buffer-surfactant mixtures. The pH optimum of the reaction was 7.5 when t-butyl hydroperoxide was the oxidant and 5.5 when hydrogen peroxide was the oxidant. With t-butyl hydroperoxide as oxidant the immobilized enzyme showed increasing activity to 65°C. The temperature profile with hydrogen peroxide was flatter, although activity was also retained to 65°C. In 1 h reactions at 25°C at their respective optimal pH values, t-butyl hydroperoxide and hydrogen peroxide promoted epoxide formation at the same rate. Larger-scale reactions were conducted using a 20-fold increase in sodium oleate (to 100 mg). Reaction time was lengthened to 24 h. At optimized levels of t-butyl hydroperoxide 80% conversion to epoxide was achieved. With hydrogen peroxide only a 33% yield of epoxide was obtained, which indicates that hydrogen peroxide may deactivate peroxygenase.
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Piazza, G.J., Foglia, T.A. & Nuñez, A. Optimizing reaction parameters for the enzymatic synthesis of epoxidized oleic acid with oat seed peroxygenase. J Amer Oil Chem Soc 78, 589–592 (2001). https://doi.org/10.1007/s11746-001-0309-9
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DOI: https://doi.org/10.1007/s11746-001-0309-9