Highly c-axis oriented zinc oxide (ZnO) thin films deposited by radio frequency (RF) magnetron sputtering, under varying ambient atmosphere (oxygen and argon reactive gas mixture), were studied for biosensing application. The as-grown ZnO thin films were found to be under compressive stress. Glucose oxidase was chosen as model enzyme for studying biosensing response properties of the ZnO thin films. The present study reveals a good correlation between stress induced during thin film growth and its biosensing response characteristic. The bio-electrodes based on ZnO thin films which are under the influence of higher stress, show better sensitivity and higher enzyme loading along with a prolonged shelf life. The study highlights the importance of physical properties of thin film matrix on its biosensing application. (C) 2015 Elsevier Inc. All rights reserved.