The ability of white-rot fungus, Trametes hirsuta AK04, to utilize phenolics as single and mixed substrates was determined in mineral medium and palm oil mill effluent (POME). The strain AK04 was able to rapidly metabolize all ten phenolics as single and mixed substrates at all test concentrations. With single substrates, between 78 and 98% removal was achieved within seven days. The biomass yield increased with increasing concentration from 100 to 500 mg L−1 but slightly decreased when the concentration was increased up to 1,000 mg L−1. When fitted to a Haldane model, the groups of benzoic and cinnamic acid derivatives gave significantly higher maximum specific growth rates than other phenolics. Phenol exhibited the lowest affinity and highest inhibitory effects on fungal metabolism. In mixed substrates, the total concentration ranges of phenolics mixtures between 1,000 and 6,000 mg L−1 did not affect the fungal growth rate and the strain AK04 showed a high degree of resistance to their toxic effects. The addition of glucose and yeast extract enhanced the degradation rates of individual phenolics in the substrate mixtures, demonstrating the advantage of this strain for treating complex media, such as industrial wastewater.