This study examined effects on shelf-life extension and prevention of starch retrogradation in rye-wheat mixed bread (RWMB) samples prepared with substitutions of 20, 40, 60, and 80% rye sourdough (RSD) as well as a control made with the addition of naturally fermented raisin extract. These effects were investigated using mold growth, water activity, and textural characteristics during a storage period of 8 days at $20^{\circ}C$. The activities of the rye sourdough were examined in terms of pH, total titratable acidity, general bacteria, lactic acid bacteria, and yeast counts. As the incubation time of the sour dough increased, pH decreased, while total titratable acidity increased. General bacteria, lactic acid bacteria, and yeast counts increased with increasing incubation time. Mold grew on the external surface of the control group after 6 days and on the surface of the RSD bread samples after 8 days, respectively. With regard to water activity, the control group had increasing activity as the storage period increased. In addition, the 20, 40, 60, and 80% RSD samples had increasing water activities until the 4th day however, these strongly decreased by the 6th day. In terms of textural characteristics, as the storage period increased, hardness increased, while springiness, cohesiveness, and resilience decreased. There were no significant differences in gumminess among the samples. The control, 20, 40, and 60% RSD samples had decreases in chewiness with increasing storage time, however, the 40% RSD sample did not differ significantly during the storage period. Finally, water activity was negatively correlated with hardness (p<0.05). In conclusion, the results indicate that substituting rye sourdough in rye-wheat mixed bread has a prolongation effect on shelf-life, but no effect on the prevention of starch retrogradation.