In this study, the effects of ozone treatment on the microorganism mortality in wheat flour and shelf-life of fresh noodles were investigated, as well as the physicochemical properties of wheat flour and textural qualities of cooked noodles. Results showed that the total plate count (TPC) can be largely reduced in wheat flour exposed to ozone gas for 30 min and 60 min. Whiteness of flour and noodle sheet, dough stability, and peak viscosity of wheat starch were all increased by ozone treatment. Free cysteine content in wheat flour was shown to decrease significantly (P<0.05) as the treatment time increased and remarkable protein aggregates were observed in both reduced and non-reduced SDS-PAGE patterns. In addition, ozone treated noodles were generally higher in firmness, springiness, and chewiness, while lower in adhesiveness. Microbial growth and darkening rate of fresh noodles made from ozone treated flour were delayed significantly.