According to the sequence of pullulanase gene from Bacillus deramificans(NCBI accession number: AX203843),the gene encoding mature peptide of pulluanase was synthesized and designated pulA.The pulA was amplified by the method of PCR and cloned into the expression vector pHY-WZX,yielding hybrid plasmid pHY-WZX-pulA.Subsequently,pHY-WZX-pulA was introduced into Bacillus licheniformis B60608.Active pullulanase was expressed by recombiant B.licheniformis and secreted into medium.Culture condition of recombinant B.licheniformis were optimized for production of pullulanase.The optimized medium consists of 2 % cotton seed protein and 8 % of glycerol.