Structure of the gene encoding laminaripentaose-producing β-1,3-glucanase (LPHase) of Streptomyces matensis DIC-108

• Published in: Journal of fermentation and bioengineering Vol. 85; no. 5; pp. 459 - 464
• Main Authors: Nakabayashi, Masahiro, Nishijima, Takashi, Ehara, Gaku, Nikaidou, Naoki, Nishihashi, Hideji, Watanabe, Takeshi
• Format: Journal Article
• Language: English
• Published: Osaka Elsevier B.V 01-01-1998; Society for Fermentation and Bioengineering
• Subjects: 3-glucanase; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; cloning; ENZIMAS; ENZYME; ENZYMES; Fundamental and applied biological sciences. Psychology; GENE; GENES; Genetics; GLUCANE; GLUCANOS; GLUCANS; laminaripentaose; Mission oriented research; NUCLEOTIDE SEQUENCE; SECUENCIA NUCLEOTIDICA; SEQUENCE NUCLEOTIDIQUE; STREPTOMYCES; Streptomyces matensis; β-1; nucleotide sequence; Streptomyces matensis; 3-glucanase; β-1; laminaripentaose; cloning; Streptomycetaceae; Purification; Nucleotide sequence; Enzyme; Sequence alignment; Homology; Gene expression; Actinomycetales; Glycosidases; N terminal-Sequence; Bacteria; Hydrolases; Actinomycetes; Glucan-endo-1,3-β-D-glucosidase; Molecular cloning; O-Glycosidases; Aminoacid sequence; Structural analysis
• ISSN: 0922-338X
• DOI: 10.1016/S0922-338X(98)80062-7

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 colony 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.