Abstract:A bacterial strain MBYD-1, capable of utilizing DMF (N, N-dimethylformamide) as sole carbon and nitrogen source, was isolated from activated sludge collected from wastewater-treating system of a pesticide manufacturer. This strain was identified as Pseudomonas sp. according to its morphological, physiological and biochemical analysis and the analysis of its 16S rRNA gene sequence, and named strain MBYD-1. The optimal pH and temperature for the degradation were 7.0 and 30℃, respectively. Strain could mineralize DMF completely in 72 hours when the initial concentration of DMF was 400 mg/L, and with 2% inoculating quantity. Degradable substrates test showed that strain MBYD-1 grew better in formate than formaldehyde, and grew better in formamide than dimethylamine.