Aims: The objective of this study was to investigate the extracellular enzymatic activity (EEA) profile of yeasts isolated from tropical environments of the Brazilian rain forest. This screening survey could constitute the first approach in selecting yeast strains of environmental origin potentially exploitable as enzyme producers. Methods and Results: In this study, 348 yeast (193 ascomycetes and 155 basidiomycetes) and 46 yeast-like strains ( Aureobasidium pullulans) were screened for their EEA profile. The spread occurrence of extracellular amylases, esterases, lipases, proteases, pectinases and chitinases appeared to be a strain-related character. Conclusions: Yeasts isolated from tropical environments could represent a promising source of EEA. Selected strains showed maximum levels of EEA under acidic or neutral conditions. Significance and Impact of the Study: This study demonstrated the potential for yeasts isolated from extreme environments as sources of industrially relevant enzymes for biotechnological purposes.