Marine-derived fungi have proven to be a prolific source of secondary metabolites with interesting structures and biological activities. Tyrosinase is a key enzyme for melanin biosynthesis and is associated with melanin hyperpigmentation and melanoma. Various culture conditions are able to produce variable bioactive secondary metabolites. Three different broth media; Potato dextrose (PD), PYMG and Czapek-Dox, in addition to one solid medium (Rice medium) were used for the cultivation of the fungus and tested for their potentiality to improve Tyrosinase inhibitory activity. The mycelia static extract of PD medium showed strong inhibitory activity (48%), followed by moderate activity for the culture static extracts of DOX and PYMG media (41 and 40%), comparing to the drug activity (vitamin C, 60%). The mycelia and culture shake of the three media, in addition to Rice solid medium extracts showed low activity. The big surprise is for PYMG culture static, mycelia and culture shake extracts, where by using distilled water instead of sea water and the reduction in glucose concentration (from 10 to 5 %) led to significant sudden increase of tyrosinase inhibitory activity from (0 to 40%). GC/MS analysis of PD extract revealed the presence of 22 compounds. The presence of benzoic acid, 2,4-dihydroxybenzoic acid, caffeic acid, benzeneacetic acid-α-4-dihydroxy, benzeneacetic acid-2-hydroxy, benzenepropanoic acid-α-hydroxy improved the level of the tyrosinase inhibitory activity. PYMG culture static extract contained 2,2’-dihydroxy-chalcone (conc. 34%) and 2,4,4’-trihydroxy-chalcone (conc. 3%), which have tyrosinase inhibitory activity.