Abstract: In November 2017, root tubers of 3-year Tetrastigma hemsleyanum were collected in Pan’an, Zhejiang province. Experiments wereconducted on extraction of polysaccharide from tubers by multienzymatic method and response surface analysis. Based on the results of single factorexperiment, enzymolysis was carried out by multienzymes (ratio of amylase, protease, pectinase of 2:2:1). Optimization of extraction technology wasimplemented on enzymatic hydrolysis temperature, time and total enzyme amount. The response surface analysis demonstrated that polysaccharideyield topped 3.84% under condition of enzymatic hydrolysis temperature of 51.7℃, total enzyme amount of 2.8%, enzymatic hydrolysis time of 92.3minutes, similar to theoretical value of 3.97%. Antioxidant activity test showed that the polysaccharide from tubers of T. hemsleyanum had the IC50 ofradical scavenging capacity on DPPH·, O2- and OH was 0.32 ,0.46,0.37 mg/mL.