This study investigated the influence of mixing and fermentation times on the chemical and physical properties of white pan bread, a typical bread consumed in Japan. Sixteen bread samples were made by combining four dough mixing stages and four dough fermentation times, and were analyzed for compression force, acids, sugars, amino acids, volatile chemicals, and overall preference in a sensory evaluation. Acids, sugars, and amino acids were analyzed using HPLC, while volatile flavor compounds were analyzed with GC/MS. The effects of mixing and fermentation times on these parameters were analyzed using analysis of variance (ANOVA). Fermentation time was positively correlated with the majority of volatile chemicals and acids, and negatively correlated with amino acids. Compression force and sugars, such as galactose and maltose, were affected by both mixing and fermentation times. Overall preference was highest in the breads mixed to the over-mixing stage and fermented for 60 min. The results of this paper are expected to be of importance in controlling the flavor, taste, and texture of white pan bread.