Cryptococcus neoformans is a pathogenic fungus that produces melanin pigmentation,which is a major virulence factor that contributes to its pathogenicity. C. neoformans strains with varying levels of melanin production were screened and selected to evaluate their growth and melanin production under different temperature ranges. This was carried out in order to ascertain the optimal conditions for melanin production and understand the effects of temperature on the phenotypic expression of C. neoformans. Out of the 150 isolates that were tested for melanin production, 10 strains of C. neoformans were isolated from three market locations in Uselu, New Benin and Egor in Benin City, Nigeria, which produced dark brown melanin pigmentation on Niger seed Agar medium.The intensity of melanin production under various temperature ranges (25 ̊C, 42 ̊C and 37 ̊C) in C. neoformans strains was also evaluated. The results showed varying intensity of very low (+), low (++), high (+++) and very high (+++++) melanin production. The optimal temperature for melanin production varied among C. neoformans strains, with some strains producing high levels of melanin at various temperatures of 42 ̊C, 37 ̊C and 25 ̊C. This research was able to demonstrate the ability of C. neoformans to produce melanin, revealing that C. neoformans strains exhibits thermotolerance and varying levels of melanin production, there is no correlation between temperature and melanin and that melanin production is not solely dependent on environmental factors but also dependent on genes.