Surfactants are amphiphatic compounds containing both hydrophilic and hydrophobic groups, which are capable of lowering surface or interfacial tension. Considering the advantages of using biosurfactants produced by microorganisms, the aim of this study was to develop and characterise a biosurfactant produced by Trichoderma sp. MK116452 isolated from Seri Cermin filling station, Brunei Darussalam, and to examine its potential application in microbial enhanced oil recovery (MEOR). The microorganism was cultured in a minimal salt medium containing 1% crude oil as a carbon source and a cell-free culture broth was used to screen its efficiency in producing biosurfactants. Characterisation of the biosurfactant showed the presence of glycosides and fatty acids, suggesting its glycolipidic nature. The isolated biosurfactant showed no toxicity to the micro-crustacean Artemia salina or to red bean ( Vigna angularis ). The extracted biosurfactant was effective at recovering up to 60% of crude oil from sand using the sand pack column method, which is similar to the recovery rate for a chemical surfactant (SDS) (65%). These findings highlight the potential use of Trichoderma sp. MK116452 biosurfactant in the oil industry.