Purification and Characterization of Melanogenic Enzyme Tyrosinase from Button Mushroom

Abstract

Melanogenesis is a biosynthetic pathway for the formation of the pigment melanin in human skin. A key enzyme, tyrosinase, catalyzes the first and only rate-limiting steps in melanogenesis. Since the discovery of its melanogenic properties, tyrosinase has been in prime focus and microbial sources of the enzyme are sought. Agaricus bisporus widely known as the common edible mushroom, it's taking place in high amounts of proteins, enzyme, carbohydrates, fibers, and low fat contents are frequently cited in the literature in relation to their nutritional value. In the present study tyrosinase from Agaricus bisporus was purified by ammonium sulphate precipitation, dialysis followed by gel filtration chromatography on Sephadex G-100, and ion exchange chromatography on DEAE-Cellulose; the enzyme was purified, 16.36-fold to give 26.6% yield on total activity in the crude extract and final specific activity of 52.19 U/mg. The SDS-PAGE electrophoresis showed a migrating protein band molecular weight of 95 kDa. The purified tyrosinase was optimized and the results revealed that the optimum values are pH 7.0 and temperature 35°C. The highest activity was reported towards its natural substrate, L-DOPA, with an apparent Km value of 0.933 mM. This indicated that tyrosinase purified from Agaricus bisporus is a potential source for medical applications.

Microbial β-Glucosidases: Screening, Characterization, Cloning and Applications

“…Other workers have used ultrafiltration [66,74], acetone precipitation [75], and ethanol precipitation [76]. The next step usually involves the use of dialysis to remove ammonium sulfate traces and other impurities present in the culture medium [77,78]. Further purification is done by chromatographic technique such as gel filtration chromatography, ions exchange chromatography, adsorption chromatography, hydrophobic interaction chromatography or HPLC [68,71,79].…”

Composition and Biological Properties of Agaricus bisporus Fruiting Bodies- a Review

“…It has been found that A. bisporus naturally contains large quantities of this compound. Tyrosinase acts as a catalyst in the hydroxylation of monophenol and the oxi-dation of diphenol in the presence of atmospheric oxygen [Kampmann et al, 2015;Labus et al, 2011;Zaidi et al, 2014]. It has also been shown that tyrosinase isolated from A. bisporus may catalyze bisphenol reactions, which could make this species a useful bioremediation agent [Kampmann et al, 2015].…”

“…This enzyme was described for the fi rst time in mammals [Bloch, 1927]. Its connection with the development of melanoma has been described, as well as its link to skin pigmentation problems (such as vitiligo or albinism) [Zaidi et al, 2014]. Tyrosinase can be extracted from various sources, i.e.…”

Study of physiologically active components in different parts of fruiting bodies of varieties of Agaricus bisporus (white mushroom)

“…Tyrosinase extracted from white mushroom shows a high degree of similarity to the one observed in human body. Thus, this species is a valuable and concurrently inexpensive source of tyrosinase [5,12,13]. The studies conducted on the enzyme isolated from A. bisporus showed that it exhibits a protective effect on the human lymphoma cell lines, protecting them from the negative effects of damaging agents such as dihydrogen peroxide [14].…”