A novel flowers-shaped porous active graphene microelectrode has been fabricated by directly electrochemical assembly and reduction of graphene oxide on platinum (Pt) wire with a diameter of 250 mu m. The flowers-shaped graphene microelectrode was characterized by SEM, XRD and electrochemical techniques. Results showed that the flowers-shaped graphene microelectrode created a favorable microenvironment for the GOD. The as prepared graphene microelectrode exhibited excellent electron transfer properties for GOD with the rate constant (k(s)) of 2.18 s(-1) and achieved the direct electrochemistry of GOD. It is notable that the glucose biosensor of GOD@flowers-shaped graphene microelectrode showed two-linear ranges in 8.0-148.0 mu M with the sensitivity of 267.34 mu A mM(-1) cm(-2) and in 148.0-430.6 mu M with the sensitivity of 38.15 mu A mM(-1) cm(-2) for detecting glucose. Furthermore, the biosensor possessed strong anti-interference, remarkable stability and appreciable reproducibility for the sensitive determination of glucose. It would also be extended to other analogous enzymes and bioactive molecules for various biological and electrochemical applications. (C) 2015 The Electrochemical Society. All rights reserved.