Peracetic acid (PAA) is widely used in sterilization, bleaching textile industry, environmental engineering, chemical synthesis, and biomimetic chemistry. A previous study reported that acetyl xylan esterase (AXE) of Bacillus subtilis CICC 20034 has high activity toward cephalosporin C and 7-aminocephalosporanic acid. In this study, we found that AXE also exhibited high perhydrolysis activity toward acetate esters and endowed itself with great industrial interest on enzyme-catalyzed preparation of PAA. Recombinant AXE of B. subtilis CICC 20034 could be efficiently produced in a low-cost autoinduction medium with an activity of 6.8 x 10(3) U/mL. The reaction conditions for the optimal synthesis of PAA were as follows: 0.30 mg/mL AXE crude enzyme, 300 mM glycerol triacetate, and 1 M hydrogen peroxide; pH 8.0, and 20 degrees C, which produced approximately 150 mM of PAA within 5 min. The AXE was then immobilized on an acrylate amino resin; the activity of the immobilized AXE was 383.7 U/g. In the presence of 1 g/mL of immobilized AXE resin, PM titer of the initial reaction batch was approximately 142.5 mM, and about 95.5 mM of PM could be produced after 10 cycles. (C) 2015 Elsevier Ltd. All rights reserved.