Abstract: Twenty-six Hyphomicrobium isolates from the sewage treatment plant and its receiving water body in Plön (Schleswig-Holstein, Germany) and two culture collection strains were screened for the occurrence of genes coding for denitrification enzymes (dissimilatory nitrate, nitrite and nitrous oxide reductases), for dinitrogen fixation (nitrogenase reductase) and for nitrification (ammonia monooxygenase catalyzing the first stage of this process) by DNA-probing. More than one half of the isolates had genes coding for denitrification enzymes. The DNA-DNA hybridization signals obtained with the gene segments correlated with enzyme activity measurements. The DNA of some isolates distinctly hybridized with the nifH probe indicating the occurrence of nitrogenase in the genus Hyphomicrobium. No signal was detected with the gene probe for nitrification. The results show that probes consisting of gene segments can be employed successfully to monitor the occurrence of genes which can show complex expression and in bacteria growing at low rates. The distribution pattern of the denitrification genes indicates that methylotrophic prosthecate bacteria of the sewage treatment plant and its receiving water body occupy different ecological niches.