[Objective] This study aimed to investigate the physical and chemical char- acteristic of Corynebacterium glutamicum in fermentation process of the glucose of wheat starch. [Method] The purity of glutamic acid in fermentation period, optical density and cell viability of bacteria were detected as indicators for regression com- parison and analysis. [Result] The relative error d=-3.316 6% within the experimental range of Warburg trace breathing apparatus and double function analyzer. The linear relationship was s1=(1-d)s2. During the fermentation process of the glucose of wheat starch, the average cell activity was 6.24 μA and the maximum cell activity was 6.61 μA. [Conclusion] Compared with optical density, cell viability can more accurate- ly reflect the physical and chemical properties of Corynebacterium glutamicum in fermentation process of the glucose of wheat starch. There was certain correlation between cell membrane phospholipids and cell viability.