Tyrosinase from agaricus bisporus

Glycerine (>99.8%), Folin–Ciocalteau reagent, and K₂S₂O₈ were obtained from Chempur (Piekary Śląskie, Poland). Ethanol (>99.8%) was obtained from Honeywell (Charlotte, NC, USA). Tyrosinase from Agaricus bisporus, 3,4-dihydroxy-l-phenylalanine (L-DOPA), 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), kojic acid (≥98.5% purity), quercetin (>95% purity), gallic acid (>97.5%), Dulbecco’s phosphate buffered saline (DPBS), Dulbecco’s modified Eagle’s medium (DMEM), bovine serum albumin (BSA), neutral red solution in DPBS (3.3 g/L) were purchased from Sigma Aldrich (Merck, Darmstadt, Germany). Fetal bovine serum (FBS) was obtained from Pan-Biotech (Aidenbach, Germany). The solvents used for phytochemical studies by HPLC-MS (water, acetonitrile, and formic acid) were purchased from Merck (Darmstadt, Germany).

Tyrosinase from Agaricus bisporus, LiP from Phanerochaete chrysosporium, veratryl alcohol (96%), veratraldehyde (99%), 4-phenoxy phenol (99%), benzyl phenyl ether (98%), dimethyl sulfoxide (DMSO, 99%), and benzyl benzoate (99%) were purchased from Sigma-Aldrich (St. Louis, USA). GGE (97%) and trimethylchlorosilane (TMCS, 1%) in N,O-bis(trimethylsilyl) trifluoroacetamide (BSTFA) [BSTFA + TMCS] were purchased from Tokyo Chemical Industry Co., Ltd (Tokyo, Japan) and Supelco analytical (Bellefonte, USA), respectively. To remove trace impurities such as metal salts, tyrosinase was solubilized in phosphate buffer (50 mM, pH 6.5) and then purified with an ultrafiltration device with a cutoff of 10 kDa (Merck Millipore, MA, USA). All other chemicals were used as the highest grade available without further purification.

Sodium phosphate monobasic monohydrate was obtained from Fluka (Buchs, Switzerland). Sodium phosphate dibasic anhydrous and methanol were obtained from Carlo Erba Reagents (Val-de-Reuil, France). Imidazole, p-nitroso dimethylaniline (RNO), and nitro blue tetrazolium chloride (NBT) were purchased from Alfa Aesar (Ward Hill, MA, USA). Tyrosinase from Agaricus bisporus, L-tyrosine, kojic acid, and DMSO were obtained from Sigma-Aldrich (St. Louis, MO, USA).