Molecular Cloning and Nucleotide Sequences of cDNA and Gene Encoding endo-Inulinase from Penicillium purpurogenumt.

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  • Molecular Cloning and Nucleotide Sequences of cDNA and Gene Encoding<i>endo</i>-Inulinase from<i>Penicillium purpurogenum</i>
  • Molecular cloning and nucleotide sequences of cDNA and gene encoding endo-inulinase from Penicillium purpurogenum

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Abstract

A cDNA and a gene encoding endo-inulinase from Penicillium purpurogenum were isolated, and were cloned for the first time. Two oligonucleotide probes, which were synthesized based on the partial amino acid sequences of the purified endo-inulinase, were used to screen a cDNA library. A 1.7-kb DNA fragment encoding endo-inulinase was isolated and analyzed. A single open reading frame, consisting of 1548-bp, was found to encode a polypeptide that comprised a 25-amino acid signal peptide and 490-amino acid mature protein. All the partial amino acid sequences of the purified enzyme were discovered in the deduced ones. The deduced amino acid sequences of endo-inulinase had similar sequences to those of fructan hydrolases. A 3.5-kb chromosomal DNA fragment encoding endo-inulinase was also isolated and analyzed. The same ORF with the cDNA clone was identified. There were no introns in the endo-inulinase gene.

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