A novel bacterial strain, HK31-GT, was isolated from a subsurface geothermal aquifer (Hellisheidi, SW-Iceland) and was characterized using a polyphasic taxonomic approach. Phylogenetic analysis of 16S rRNA gene along with phylogenomic position indicated that the novel strain belongs to the genus Phenylobacterium. Cells are motile Gram-negative bacilli. Physiological characterization showed that strain HK31-GT is a mesophilic bacterium able to grow from 10 to 30 °C, at pH values between 2 and 12 and at NaCl concentrations between 0 and 0.5%. Optimal growth was observed without sodium chloride at 25 °C and pH 6. Strain HK31-GT is chemoorganoheterotroph and its major saturated fatty acids are C18:1ω7c, C16 :1ω6c and C16:0, the predominant quinone is Q-10 and the major polar lipid is phosphatidylglycerol. The new strain also possesses the capacity to use ferrous iron (Fe(II)) as the sole energy source and can also be considered as a chemolithoautotrophic microorganism. The overall genome of strain HK31-GT was estimated to be 4.46 Mbp in size with a DNA G+C content of 67.95%. Genes involved in iron metabolism were identified, but no genes typically involved in Fe(II) oxidation were found. Average nucleotide identity and digital DNA-DNA hybridization values between the genome of strain HK31-GT and the genomes of its closest relatives are below the species delineation threshold. Therefore, given the polyphasic approach used, strain HK31-GT represents a novel species of the genus Phenylobacterium, for which the name Phenylobacterium ferrooxidans sp. nov. is proposed. The type strain is HK31-GT (DSM 116432T = UBOCC-M-3429T= LMG 33376T).