Biochar is a by-product from the pyrolysis of biomass and has a great potential in soil amendment due to its carbon and nutrient-rich properties. The aim of this study was to investigate the impact of increasing amounts (0, 0.01, 0.1, 0.2, 0.5 and 1.0%) of two types of biochar (so-called enhanced and non-enhanced) to soil on the biodegradation of 14C-phenanthrene. Enhanced biochar contains inoculants which are designed to potentially stimulate microbial activity and promote biological function in soil. After 100 d of incubation, the addition of 0.5% and 1% enhanced (EbioC) and non-enhanced biochars (NEbioC) led to longer lag phases, reduced rates and extents of 14C-phenanthrene in amended soil. However, in soils amended with 0.01%, 0.1% and 0.2% amendments, extents of mineralisation of 14C-phenanthrene increased and were found to be higher in the EBioC- as compared to the NEbioC-amended soils. Increasing soil-phenanthrene contact time also increased 14C-phenanthrene mineralisation in soil which had received smaller amounts of EBioC. Application of both EbioC and NEbioC also enriched the soil microbial populations during the incubation. However, it was found that phenanthrene-degrading microbial populations declined as soil contact time increased; this was particularly true for soils receiving larger amounts of due to reduction in the mobile/bioaccessible fraction of the phenanthrene in soil. The findings revealed the importance of the type and amount of biochar that may be added to soil to stimulate or enhance organic contaminant biodegradation.