Abstract
Blastocystis sp. is a commonly encountered gut protozoan with unclear pathogenicity. The presence of proteases in this organism may be related to its potential pathogenicity or other physiological differences. This study aimed to identify the various proteases that may be present in different subtypes (STs) of Blastocystis sp. using azocasein assay and gelatin zymography. In this study, cysteine, serine, aspartic protease, metalloproteases, and unknown proteases were identified in Blastocystis sp. cultures obtained from animal and water samples belonging to ST1-ST5 and ST7. Azocasein assay and gelatin zymography were conducted on different batches of protease extracts, which showed varying results. Cysteine protease was the most commonly encountered. Protease activity in general is not associated with ST or source (animal or water). However, the presence of cysteine protease alone was associated with animal samples, whereas the presence of more than two protease types was associated with water samples. Azocasein assay and gelatin zymography were conducted on different batches of protease extracts, which showed varying results. Protease activity and the types present may change over time. Blastocystis sp. shows high protease diversity, including possible novel types.