An effective electrochemical sensor for myoglobin (Myb) detection was developed using a simple procedure of denaturing the protein with guanidine hydrochloride and detecting the released heme group by cyclic voltammetry. The concentration of denaturant was optimized to obtain maximum current response for the analyte (Myb). To improve the sensitivity of the sensor, the working electrode, glassy carbon electrode was modified with a layer of Titania nanotubes (TNT). The direct electrochemical behavior of the modified electrode (TNT-GCE) was studied using cyclic voltammetry (CV). The performance of the sensor was investigated and optimized and the system was evaluated by monitoring the Myb concentration. Despite the reduced current response for the modified electrode compared to bare GCE, the sensitivity of the system was improved significantly by overcoming the large background current due to denaturant. The developed TNT modified electrode improved the detection limit of Myb and showed good stability, sensitivity and reproducibility. Under optimal conditions, the catalytic currents are linearly proportional to the concentrations of Myb in the wide range from 50 nM to 6 M. This approach provides improved sensitivity in the given range, and may provide a novel and efficient platform for the fabrication of sensors for other heme proteins.