In this study, the ability of 4-n-nonylphenol (4-n-NP) elimination by fungal species belonging to the genus Metarhizium was investigated. The occurrence of 35 metabolites from 4-n-NP degradation was confirmed. For the first time, based on the obtained results, the 4-n-NP biodegradation pathway distinctive for the genus Metarhizium was proposed. Principal Component Analysis (PCA) indicated that despite the similar elimination pathway in all the examined Metarhizium species, there are significant differences in the kinetics of degradation of 4-n-NP. Oxidation of the terminal methyl group of the aliphatic chain leading to the formation of carboxylic acids coupled with the removal of terminal carbon is characteristic of M. robertsii and M. guizhouense, whereas metabolites with a hydroxyl group in the distal part of the nonyl chain distinguish M. lepidiotae and M. majus. Additionally, this study verified the participation of cytochrome P450 in the elimination of the xenobiotic by Metarhizium as experimentally proven for M. robertsii.