Oil pollution, especially in the marine environment, has become a serious environmental problem. Hexadecane (HXD) is a major alkane component and it is present in the aliphatic fragment of crude oil, which can be used by fungi as a sole carbon source. Biosurfactant which is produced by fungi facilitates HXD degradation. This study investigated the ability of mangrove fungi to be used as HXD and produce biosurfactant. The medium used to determine the ability of fungi to use hexadecane is MSM-HXD 2%, whereas Hua medium is used for determining the potential for producing biosurfactants. Biosurfactant production by fungi strains was indicated by oil displacement test, the reduction in surface tension and emulsion index. The results showed that 13 out of 16 species of fungi can use HXD as a sole carbon source. Inonotus radiatus LM 3020 is observed to be the most capable isolate to use HXD with a growth ratio of 6.0, and can produce biosurfactant with a positive oil displacement test (ODT) value whose minimum surface tension was 54.01 dyne/cm but the emulsion index was found to be zero.