Extraction, purification of tyrosinase and tyrosinase inhibitory activity of four medicinal plants from Nanded district (MS), India

Abstract

The systematic study aims to explore the tyrosinase inhibitory activity of selected plants. The study included extraction, purification, and inhibition of the enzyme. Tyrosinase has an important role in melanin formation, is responsible for the production of colour pigments of skin, hair, and eye. In the presents study, tyrosinase was isolated from Mushrooms, isolation of enzyme was done by acetone precipitation procedure and precipitation of enzyme was done with ammonium sulphate precipitation method. Plants selected for extraction were Azadirachta indica (Neem), Manikara zapota (Chiku), Annona squamosa (Sitaphal), and Hibiscus Rosa-sinesis (China rose). For phytochemical screening Alkaloids-Mayer’s Test, Flavonoids (Shinoda Test, Alkaline Reagent Test), sugar (Benedict’s reagent Test), Glycosides (Borntrager's Test), Phenolic compounds Test (Ferric chloride Test, Gelatin Test, Lead Acetate Test). Mushroom tyrosinase inhibitory assay was determined by the spectroscopic method. The enzyme precipitates on 85% saturation of ammonium sulphate, dialyzed and loaded on preequilibrated DEAE cellulose column for further purification at 1ml/min flow rate. Purification shows the 26 frac-tions giving very good enzyme activity. The specific activity of the enzyme after all purification steps was found to be 2 U/µg proteins with 17.09- fold purify-cation.