In order to verify the cryoprotective effect of an antifreeze protein (BaAFP-1) obtained from barley on bread dough, the effect of BaAFP-1 on the rheological properties, microstructure, fermentation, and baking performance including the proofing time and the specific volume of bread dough and bread crumb properties during freezing treatment and freeze-thaw cycles were analysed. BaAFP-1 reduced the rate of decrease in storage modulus and loss modulus values during freezing treatment and freeze-thaw cycles. It influenced the formation and the shape of ice formed during freezing and inhibited ice recrystallization during freeze-thaw. BaAFP-1 maintained gas production ability and gas retention properties, protected gluten network and the yeast cells from deterioration caused by ice formation and ice crystals recrystallisation in dough samples during freezing treatment and freeze-thaw treatment. It slow down the increase rate of hardness of bread crumb. The average area of pores in bread crumbs decreased significantly (p < 0.05) as the total number of pores increased (p < 0.05), and the addition of BaAFP-1 inhibited this deterioration. These results confirmed the cryoprotective activity of BaAFP-1 in bread dough during freezing treatment and freeze-thaw cycles.