During experiments in animal studies, it has been observed that enamelysin (MMP-20) is expressed during tooth development in the late secretory stage of amelogenesis but not in the mature enamel. The aim of this research was to determine the location of MMP-20 in human tooth germs in the different structures of the enamel organ. The detection of MMP-20 was performed by immunohistochemistry in 20 specimens obtained from human fetuses. Immunostaining of MMP-20 was observed from the presecretor stadium in stellate reticulum and intermediate stratum and in the basal portion of ameloblasts in the secretory stage in stellate reticulum, stratum intermedium, secretory ameloblasts, odontoblasts and dental papilla. The results of this research show the location of MMP-20 in tooth germ development in humans and provides the foundation for future research about the process of dental organ formation.