A polychlorophenol-degrading strain, designated MT1T, and three MT1-like strains, MT101, MT103 and MT104, were isolated from a cold (4–8 °C) fluidized-bed process treating chlorophenol-contaminated groundwater in southern Finland. The organisms were Gram-negative, rod-shaped, catalase-positive, non-spore-forming and non-motile. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strains belonged to theα-4 subclass of theProteobacteriaand were members of the genusNovosphingobium. The highest 16S rRNA gene sequence similarity observed for these strains was 96·5 % with the type strains ofNovosphingobium hassiacum,Novosphingobium aromaticivoransandNovosphingobium subterraneum. Chemotaxonomic data (major ubiquinone: Q-10; major polyamine: spermidine; major polar lipids: phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine and sphingoglycolipid; major fatty acids: 18 : 1ω7c, 16 : 1ω7cand 2-OH 14 : 0) as well as the ability to reduce nitrate supported the affiliation of the strains to the genusNovosphingobium. Based on the phylogenetic analysis, whole-cell fatty acid composition as well as biochemical and physiological characteristics, the MT1-like strains were highly similar and could be separated from all recognizedNovosphingobiumspecies. The novel speciesNovosphingobium lentumsp. nov. is proposed to accommodate strains MT1T(=DSM 13663T=CCUG 45847T), MT101 (=CCUG 45849), MT103 (=CCUG 45850) and MT104 (=CCUG 45851).