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Cloning and Sequence Analysis of a GH10 β-1,4-Xylanase Gene of Cellulosimicrobium cellulans DY-8 Isolated from the Gut of Moechotypa diphysis
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- URLhttps://db.koreascholar.com/Article/Detail/288460
A xylan-decomposing Gram-positive bacterium, Cellulosimicrobium cellulans DY-8, was isolated from the gut of a wood-feeding longicorn beetle, Moechotypa diphysis. To amplify a partial fragment of the GH 10 β-1,4- xylanase (XylC) gene of strain DY-8, two degenerated oligonucleotide primers were designed based on strictly conserved regions (WDVVNE and ITELLDV) in the GH family 10 xylanolytic enzymes. The full gene (1488-bp) of XylC, which was predicted to encode a protein consisting of 495 amino acids with a molecular mass of 52.0 kDa and a calculated pI of 6.49, was cloned by repeated DNA walking and nested PCR protocols. The results of a protein blast survey showed that XylC was a β -1,4-xylanase comprised of an N-terminal catalytic GH10 domain (from Ser48 to Leu338) and a C-terminal RICIN domain (from Tyr359 to Leu492). This overall structure of XylC was 57% identical to that of Actinoplanes sp. SE50/110 β -1,4-xylanase (Accession number: YP_006265966), which has not yet been biochemically characterized.
