We present an easy and fast way to fabricate glucose oxidase-diphenylalanine-reduced graphene oxide (GOx-FF-rGO) film by using pulsed potentiostatic method. Glassy carbon electrode (GCE) modified by the obtained GOx-FF-rGO nanocomposite film is systematically characterized for its application in glucose detection. Under optimized condition, the biosensor exhibits a wide linear response in the range from 0.1 to 30.0 mmol L-1, with a low detection limit down to 0.01 mmol L-1. Our results indicate that the pulsed potentiostatic deposition approach may lead to production of highly sensitive and stable electrochemical biosensors. (C) 2017 Elsevier Ltd. All rights reserved.