Abstract: In order to select the traditional starters suitable for baking dough fermentation and improve the flavor and quality of pancakes, 10 traditional starter cultures were collected from different regions including Nanyang (NY), Xinxiang (XX), Zhoukou (ZK), Luohe (LH), Rizhao (RZ), Zhumadian (ZMD), Heze (HE), Tai’an (TA), Zhengzhou (ZZ) and Weinan (WN). The basic characteristics and the effects on microbial quantity, reducing sugar content during dough fermentation and flavor quality of pancakes were studied. The results showed that the pH values of traditional starter from different regions ranged from 4.26 to 5.29, with TTA values between 8.18 mL and 17.49 mL. Significant differences were observed in the colony counts of yeast and lactic acid bacteria. The yeast and lactic acid bacteria in traditional starters from RZ and LH exhibited relatively strong acid and alcohol production capabilities. Meanwhile, the yeast and lactic acid bacteria in the traditional starter showed similar growth trends in the dough fermentation, most of them with rapid growth in the early stage and a slowing down and stabilization in the later stage. However, the traditional starters from RZ and XX demonstrated better fermentation capacities in the early fermentation stage. The traditional starter from RZ had the fastest fermentation rate in the early 2 h of fermentation, while the traditional starters from XX performed the best fermentation power between 2 and 4 h of fermentation. Compared with angel yeast, the specific volume of pancakes prepared by XX, LH and RZ starters showed no significant difference, but the hardness and chewiness were significantly reduced, and the sensory scores of pancakes prepared with XX, LH and RZ starters were at the highest levels, with scores of 87.60, 86.40, and 86.20, respectively. In conclusion, the traditional starters from XX, LH and RZ contribute to enhancing the flavor and quality of pancakes.