(S)-2-aminobutyramide is the key chiral intermediate of levetiracetam, an important drug for the treatment of epilepsy. A recombinant amidase Dt-Ami 6 was screened as the best biocatalyst for enantioselective synthesis of (S) -2 -aminobutyramide. The reaction parameters including temperature, pH, organic cosolvent and substrate concentrations were investigated. The results indicated that the optimum temperature, substrate concentration, biocatalyst loading and reaction time were 40 ℃, 1 mol/L, 8 g/L and 40 min, respectively. Under these conditions, (S)-2- aminobutyramide was obtained in 41.7% yield and 94.0% e.e..