Continuous use of the pyrethroid insecticide beta-cypermethrin (beta-cp) has resulted in serious environmental contamination problems. We report here that a novel bacterial strain BSF01, which was isolated from activated sludge and identified as Bacillus subtilis (collection number: CCTCC AB 2014103), showed high efficiency in degrading beta-cp. Strain BSF01 was able to utilize beta-cp as the sole carbon source for growth and degraded 89.4 % of 50 mg L−1 beta-cp within 7 days. The optimal conditions for beta-cp degradation were determined to be 34.5 °C, pH 6.7, and inocula amount 0.11 g dry wt L−1 using response surface methodology. The kinetic parameters qmax, Ks, and Ki were established to be 2.19 day−1, 76.37 mg L−1, and 54.14 mg L−1, respectively. The critical inhibitor concentration was determined to be 64.30 mg L−1. Seven metabolites were identified by gas chromatography–mass spectrometry. Furthermore, a novel biodegradation pathway for beta-cp was proposed on the basis of analysis of the metabolites. This strain was also capable of degrading a wide range of pyrethroid insecticides including cypermethrin, deltamethrin, cyhalothrin, and beta-cyfluthrin, which similar to beta-cp are hazardous chemicals. Taken together, our results depict the biodegradation pathway of beta-cp and highlight the promising potentials of strain BSF01 in bioremediation of pyrethroid-contaminated environments.