Purified Mushroom Tyrosinase Induced Melanogenic Protein Expression in B16F10 Melanocytes: A Quantitative Densitometric Analysis

Abstract

Background: Human skin exists in a wide range of different colors and gradations, ranging from white to brown to black. This is due to the presence of a chemically inert and stable pigment known as melanin, which is produced deep inside the skin but is displayed as a mosaic at the surface of the body. Methods & Materials: In mammalian melanocytes, melanosome is a highly specialized organelle where melanin is synthesized. Melanin synthesis is controlled by tyrosinase, the vital enzyme in melanogenic pathway. The present investigation is based on the effect of purified tyrosinase of Agaricus bisporus on B16F10 melanocytes for melanogenic protein expression. Results: After the treatment of purified tyrosinase B16F10 melanocytes did not show any cytotoxic effect. Melanin content in B16F10 melanocytes was increased by purified tyrosinase in a dose-dependent manner. Quantitative western blot analysis revealed that cellular tyrosinase intensity was enhanced after treatment with purified tyrosinase for 48 hours, where the band intensity had a steady increase in the absorption of purified tyrosinase in B16F10 cells. The density analysis described increased absorption for 2 to 5 bands as 2.7, 3.7, 6.7 and 8.6% respectively. The bands in the comparative analysis of western blot were between the Rf value range (0.40-0.57) with maximum absorption of 3000 intensity curve at 32μg/mL, rather than higher concentration 64μg/mL, showing a decrease in the absorption. Conclusion: It is presumed that purified tyrosinase can be used as contestants for the treatment of vitiligous skin conditions.