This study describes a novel biosensor method for specific determination of nitrate in food and water samples by using nitrate reductase (NR) (EC 184.108.40.206) biosensor based on the detection of oxidation peak current of redox mediator, methyl viologen, related to nitrate concentration. The method was shown to be selective and sensitive to determine the nitrate levels of water samples and processed meat samples. Immobilization procedure and also working conditions of the biosensor were optimized. Dynamic range attained with this method was established as (5.0-90.0x10-9M) for nitrate concentration with a 10s response time. Limit of detection (LOD) and quantification (LOQ) of the biosensor were calculated as 2.2x10-9M and 5.79x10-9M, respectively. Reproducibility experiments was established on repetitive measurements by using a freshly prepared biosensor for avoiding the memory effect. The RSD was calculated as 1.22% at a nitrate concentration of 4.7x10-8M (n=7).