The purification of recombinant Pfu DNA polymerase expressed in Escherichia coli was studied. The lysed supernatant was heated to 75 °C to denature E. coli protein, followed by chromatography on JK110 and Sephadex G-75. The purified protein had comparable activity to the commercially obtained Pfu in both DNA polymerase and PCR amplification. The final product had a specific activity of 17,600 U/mg and 149,600 U of Pfu DNA polymerase was obtained from 500ml culture. JK110 has worked well in our study and appears to be a new method of choice for purification of Pfu DNA polymerase.