Structure of the Gene Encoding Laminaripentaose-Producing β-1, 3-Glucanase(LPHase)of Streptomyces matensis DIC-108

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Streptomyces matensis DIC-108 secretes a laminaripentaose-producing β-1, 3-glucanase(LPHase)into the culture medium. LPHase was partially purified and its N-terminal amino acid sequence was determined. The gene(lph)for LPHase was cloned using a cology hybridization technique and a synthetic oligonucleotide probe designed from the N-terminal amino acid sequence, and the nucleotide sequence of lph was determined. lph has an open reading frame of 1203 bp that encode a polypeptide of 401 amino acids including a putative 35-amino acid signal sequence. The mature portion of the polypeptide showed significant sequence similarity with the catalytic domains of β-1, 3-glucanases of Oerskovia xanthineolytica and Arthrobacter sp. strain YCWD3. Since lph was not expressed in Escherichia coli, the coding region of lph was amplified by PCR and ligated into the expression vector pKK233-2. E. coli JM109 cells carrying the resultant plasmid pKK-lph produced β-1, 3-glucanase of a size identical to that of LPHase produced by S.matensis DIC-108, and the enzyme exclusively liberated laminaripentaose from insoluble β-1, 3-glucan.
社団法人日本生物工学会の論文
1998-05-25