Saccharomyces cerevisiae, Leuconostoc lactis and Lactobacillus plantarum, which were screened from grape natural fermentation broth, were used to prepare an excellent natural mixed starter to improve the baking quality of bread. The optimal ratio of three strains in the starter was 1∶1.5∶1.5, meanwhile, the specific volume and hardness of the bread were better than those of the common yeast bread. The texture analyzer (TA), differential scanning calorimetry (DSC) and solid phase micro-extraction-gas chromatographic-mass spectrometry (SPME-GC-MS) were used to determine the aging properties and flavor of bread fermented by natural mixed starter, and the results showed that the natural mixed starter could reduce the hardness and aging enthalpy of bread during storage and delay the staling of the bread, thus the number and the relative content of volatile flavor compounds in bread increased by 11 species and 4.81%, respectively. The survival rate of natural mixed starter was 81.78% after 90 days of storage at 4 ℃. The starter with the advantages of high viability and fermentation ability could be applied in bakery industry to improve quality, flavor and shelf life of bread.