The aim of this study was to prepare a function bread by adding mulberry fruit powder, and investigate the influence on physical, sensorial and biological properties of the bread. The response surface optimization experiments were used to optimize the formulation of mulberry enriched bread. The contents of total polyphenols, total flavonoids and total polysaccharides in mulberry bread were determined by UV spectrophotometry. The antioxidant activity and enzyme activity inhibition properties of mulberry bread were also determined. The results showed that the highest sensory scores of mulberry breads were obtained when mulberry fruit powder, butter, yeast and salt were added at 11.3%, 5.8%, 1.2% and 0.9%. The total polyphenols, flavonoids and polysaccharides were 1.63 ± 0.01 mg/g, 1.05 ± 0.06 mg/g and 11.72 ± 0.1 mg/g. The DPPH radical scavenging, OH•ˉ radical scavenging and ABTS radical scavenging capacities of mulberry bread were 66.83 ± 1.23%, 48.53 ± 0.87% and 51.24 ± 1.51%. The α-amylase and α-glucosidase enzyme inhibition rate of mulberry bread was 15.86 ± 1.24% and 18.91 ± 1.87%, respectively. In vitro digestive experiments had shown that the addition of mulberry fruit powder could reduce the amount of glucose releasing, and slowing down the rise of postprandial blood sugar in humans. Mulberry fruit addition in bread production enables the formulation of high-nutritive, good antioxidative, low-energy bread; and enzyme activity inhibitory properties.