Abstract: A new nitrilase producing strain showing high hydrolyzing activity towards 3-cyanopyridine was isolated from the soil using enrichment technique. Microorganisms were enriched with glycerol as the carbon source and nitriles as nitrogen source. Two different nitriles were used for screening, and it was found out and confirmed that 8 strains isolated dependent on acetonitrile as nitrogen source, can produce arylacetonitrilases, two of which showed 3-cyanopyridine hydrolysing activity; on the other hand, another two strains, compared with those former strains, show exclusive identification towards 3-cyanopyridine, as well as producing aromatic nitrilase when served with 3-cyanopyridine as nitrogen source. The one showing the higher activity towards 3-cyanopyridine is identified as Rhodobacter sp., the capacity of which for producing nitrilase was first reported.