Acetyl esterase was produced by a bacterial strain RB3 at a level of 0.59 U mL(-1). The strain was isolated from beef cattle rumen fluid under anaerobic condition, and was identified as Escherichia coli. The peak activity of the enzyme appeared after 48 h of culturing under anaerobic condition. The optimal pH of the enzyme activity was 8.0, and the optimal temperature was 40 degrees C. The K-m and V-max values on p-nitrophenyl acetate were 0.84 mM and 0.13 mmol p-nitrophenol liberated min(-1) mg of protein(-1) respectively. The enzyme activity could be promoted by Zn2+, Ni2+, Fe2+, and K+, and inhibited by Cu2+, Fe3+, Mn2+, Mg2+, Ca2+, and Co2+. Biodegradation of rice stalk and maize stover by the strain RB3 and Pleurotus ostreatus was compared. The strain showed higher degradation rate for hemicellulose in the crop residues, while P. ostreatus showed higher degradation rate for cellulose. This indicated the potential industrial application of the strain RB3, particularly in utilizing renewable lignocellulose containing acetyl xylan for fermentation of products. (C) 2014 Elsevier Ltd. All rights reserved.