<b>Objectives</b><br/> In an attempt α-amylase gene from <i>Pyrococcus woesei</i> was amplified and cloned into a pTYB2 vector to generate the recombinant plasmid pTY- α-amylase.<br/><b>Methods</b><br/> <i>Escherichia coli</i> BL21 used as a host and protein expression was applied using IPTG. SDS-PAGE assay demonstrated the 100 kDa protein. Amylolytic activity of proteins produced by transformed <i>E. coli</i> cells was detected by zymography, and the rate of active α-amylase with and without the intein tag in both soluble conditions and as inclusion bodies solubilized by 4M urea were measured.<br/><b>Results</b><br/> Amylolytic activity of ∼185,000 U/L of bacterial culture was observed from the soluble form of the protein using this system.<br/><b>Conclusion</b><br/> These results indicate that this expression system was appropriate for the production of thermostable α-amylase.