The program warming method was applied to carry out the enzymatic hydrolysis of highland barley. The response surface methodology was used to optimize the enzymatic hydrolysis process of probiotic fermented barley with the reducing sugar content as the response value. The result indicated that the optimal conditions could be gotten: with the ratio of solid to liquid 115(g/mL) to dissolve highland barley powder, pasting and cooling to 45 ℃, addition of 8.9 U/g α-amylase, 2.0% glucoamylase, 3.5% bromelain, incubating for 20 min and then heating to 60 ℃, incubating for saccharification to the iodine test showing colorless, getting highland barley enzyme saccharification liquid by filtration. Under the condtion, the content of reducing sugar was up to 5.68% and the content of amino acid nitrogen was 0.47 g/L, which could be used to produce probiotic fermented highland barley beverage.