The Effects of dynamic high pressure microfluidization (DHPM) combined with Maillard reaction on structure and properties of α-lactalbumin (α-LA) were investigated by SDS-PAGE and spectroscopy. The results demonstrated that the fluorescence intensity and surface hydrophobicity (H₀) of α-LA treated by DHPM firstly increased then decreased, with the maximum values at 110 MPa. After Maillard reaction (MR) with lactose, the fluorescence intensity of α-LA was declined. However, H₀, emulsibility and anti-oxidation activity of α-LA was improved. These results illustrated that tertiary structure of α-LA was changed due to DHPM pretreatment, thereby promoting MR, emulsibility and anti-oxidation activity of Maillard reaction products. DHPM pretreatment combined with MR was an effect method for protein modification.