Shiitake (Lentinula edodes [Berk.] Pegler) stipe was used to substitute wheat flour to make bread. Bread quality including specific volume, color and sensory evaluation, and taste components including proximate composition, soluble sugars, free amino acids and 5′-nucleotides in shiitake stipe bread were analyzed and compared with those of white bread. Specific volumes were 4.02, 4.00, 3.89 and 3.75 cm3/g for white bread, 2%, 5% and 7% shiitake stipe breads, respectively. All breads had a comparable profile in proximate composition. Furthermore, 5% shiitake stipe bread contained more total soluble sugars and total free amino acids. Both breads contained considerately low amounts of monosodium glutamate-like and sweet components of free amino acids. Content of flavor 5′-nucleotides in 5% shiitake stipe bread was much higher than that in white bread. Shiitake stipe flour added into the dough resulted in 5% shiitake stipe bread with lower lightness and whiteness index values. All sensory results exhibited that substituting 5% of wheat flour in the bread formula with shiitake stipe flour did significantly lower bread acceptability. Overall, shiitake stipe could be incorporated into bread to provide its beneficial health effects.