An efficient method for the production of 4-cyanobenzoic acid from terephthalonitrile was developed by using a novel nitrilase from Pantoea sp. AS-PWVM4 (PaNIT). Under the optimized reaction conditions (pH 8.0 and 37 degrees C), up to 100 g/L substrate concentration was almost completely converted to 4-cyanobenzoic acid with wet whole cells loading of 5.0 wt% in 4 h and the isolated yield was 91%. The substrate concentration, yield and productivity (26.13 g/(L h)) are much higher than those in the previous studies. Furthermore, PaNIT has broad substrate specificity of catalyzing the hydrolysis of various aromatic nitriles and aliphatic nitriles. These results suggest that PaNIT is a valuable nitrilase with potential practical applications.