Cloning and Nucleotide Sequence of the Pullulanase Gene of Thermus thermophilus HB8 and Production of the Enzyme in Escherichia coli

抄録

A 3.4-kb SphI fragment carrying the pullulanase gene of Thermus thermophilus HB8 was cloned. Based on the nucleotide sequence of it and the flanking region analyzed by direct sequencing of the inverse PCR product, an expression vector was constructed. The E. coli cells harboring the plasmid produced an about 80-kDa protein having pullulanase activity, the optimum temperature of which was 70°C.