Formation of Anethole Dimer Compound Using Biocatalyst Laccase from White Oyster Mushroom

Abstract

Due to their specificity and environmental friendliness, enzymes are frequently used as biocatalysts for organic compound synthesis. The goal of this research is to synthesize anethole dimer using the laccase enzyme as the biocatalyst. Laccase is a widely used enzyme that belongs to the oxidoreductase group. This study employed anise oil containing 90% anethole, laccase isolated from the white oyster mushroom (Pleurotus ostreatus) with an activity of 712,758 U/L, and hydroquinone as a mediator. The anethole dimerization was conducted in a biphasic medium (ethyl acetate and phosphate buffer in a 4:1 ratio) for 24 and 48 hours. The reaction mixture was extracted with ethyl acetate, yielding a brownish, viscous liquid that exhibited increased color intensity after 48 hours. Gas chromatography analysis of anise oil and the reactions after 24 and 48 hours revealed increased peak intensity and changes in peaks after 48 hours. The percentage area of anethole and p-anisaldehyde was smaller in the 48-hour reaction than in the 24-hour reaction. Identification of the compounds after 48 hours suggested the formation of new compounds resulting from oxidation reactions. Examples of these new compounds include caryophyllene oxide; however, the dimerization of the anethole compound was not conclusively identified.