Abstract: Cassava resistant starch (RS3) was prepared by thermostable α-amylase and pullulanase. The RS content, the amylose contents, freeze-thaw stabilization and water-holding capacity of the RS were determined and compared between the α-amylase-RS and the pullulanase-RS. The results indicated that the α-amylase-RS content was ranging in 9. 4% ～ 12. 4%, the amylose content decreased with the increasing of the acting time. The higher-amylose-content-RS might have lower freeze-thaw stability. The water-holding capacity of it was ranging in 3. 7 ～ 5. 8g/g. The pullulanase-RS content was ranging in 4% ～ 7. 9%, the amylose content did not necessarily increase with the increasing of the acting time. The higher-amylose-content-RS may have higher freeze-thaw stability and water-holding capacity. Compared with the pullulanase-RS, the α-amylase-RS had higher RS content, higher freeze-thaw stability but lower water-holding capacity.