This paper reports covalent immobilization of xylanase by response surface methodology on glutaraldehyde-activated aluminum oxide pellets. The optimization of process parameters gave an immobilization yield of 83.65%. The bound enzyme displayed increase in optimum temperature (from 50 to 60 degrees C) and V-max (from 3333.33 to 5000 IU/mL) in comparison with free enzyme. The pH and temperature stability were also enhanced. An observed increase in half-lives and D-values resulted in improved thermostability. Thermodynamically, increase in enthalpy and free energy change after covalent immobilization could be credited to the enhanced stability. Immobilized xylanase could be reused for 10 consecutive cycles retaining 60% of its initial activity. It was found to be effective in releasing reducing sugar from poultry feed. Immobilization on aluminum oxide pellets is important due to their mechanical resistance at high pH and temperature. So, considerable stability and reusability of bound enzyme may be advantageous for its industrial application. (C) 2012 Elsevier Ltd. All rights reserved.