Biological control has been shown to be one of the possible biotechnological applications of fungi and their proteases. The objective of this study was to evaluate the nematicidal activity of the fungus Pleurotus ostreatus and its proteases on Panagrellus sp. larvae. Proteolytic activity of P. ostreatus (PLO 06) was measured and characterized at different pHs, temperatures and in the presence of a inhibitor (PMSF). Daily samples of culture medium were collected in order to determine the period of maximum enzyme production. A zymogram showed the profile of several proteases. Predatory activity of the fungus P. ostreatus (PLO 06) was evaluated on Panagrellus sp. larvae (assay A) as well as the nematicidal activity of PLO 06 proteases on the same larvae (assay B). At pH 9 and 60°C, the activity of the proteases reached the maximum. In the presence of inhibitor, there was no proteolytic activity. A sample collected on the fifth day of incubation showed the highest enzyme activity. P. ostreatus demonstrated capture activity on larvae Panagrellus sp. The values of the reduction of the larvae (Assay A) were: day 1 (65.6%); day 2 (77.4%); day 3 (95.2%). The reduction of the larvae (Assay B) was 42%. P. ostreatus (PLO 06) and its proteases were very effective against Panagrellus sp. larvae, demonstrating great potential for use in integrated biological control.