With glucose as co-substrate, anaerobic degradation kinetics of 2,4-dinitrophenol (2,4-DNP) were investigated in batch culture. The results show that 2,4-DNP and glucose can be degraded by the bacteria simultaneously. Although the effect of COD on 2,4-DNP degradation was minimal, addition of 2,4-DNP effected COD degradation obviously. The rate of 2,4-DNP degradation increased with increasing initial 2,4-DNP concentrations up to 225 mg/L. Further increase in initial 2,4-DNP concentrations caused decrease in the rate of degradation because of substrate inhibition. Uncompetitive inhibition equation is proposed to describe the degradation of 2,4-DNP. With non-linear regression technology, the kinetic model parameter q(max), K(s) and K(i) are found to be 3.24 mg/(h x g), 196.23 mg/L and 165.91 mg/L respectively. The experimental data verification for the model equation is satisfactory.