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Volume 146, Issue 11

A novel thermostable multidomain 1,4-β-xylanase from ‘’ and effect of its xylan-binding domain on enzyme activity Free

Abstract

The GenBank accession number for the sequence reported in this paper is AF200304.

The nucleotide sequence of the complete gene, encoding a novel multidomain xylanase XynA of ‘’, was determined by genomic-walking PCR. The putative XynA comprises an N-terminal domain (D1), recently identified as a xylan-binding domain (XBD), homologous to non-catalytic thermostabilizing domains from other xylanases. D1 is followed by a xylanase catalytic domain (D2) homologous to family 10 glycosyl hydrolases. Downstream of this domain two cellulose-binding domains (CBD), D3 and D4, were found linked via proline-threonine (PT)-rich peptides. Both CBDs showed sequence similarity to family IIIb CBDs. Upstream of an incomplete open reading frame was identified, encoding a putative C-terminal CBD homologous to family IIIb CBDs. Two expression plasmids encoding the N-terminal XBD plus the catalytic domain (XynAd1/2) and the xylanase catalytic domain alone (XynAd2) were constructed and the biochemical properties of the recombinant enzymes compared. The absence of the XBD resulted in a decrease in thermostability of the catalytic domain from 70 °C (XynAd1/2) to 60 °C (XynAd2). Substrate-specificity experiments and analysis of the main products released from xylan hydrolysis indicate that both recombinant enzymes act as endo-1,4-β-xylanases, but differ in their ability to cleave small xylooligosaccharides.

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Keyword(s): CBD, cellulose-binding domain , hemicellulose degradation , PT-linker, proline-threonine linker , thermophilic bacteria , thermostabilizing domain , TSD, thermostabilizing domain , XBD, xylan-binding domain , XU, xylanase unit and xynA
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2000-11-01
2025-03-17
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/content/journal/micro/10.1099/00221287-146-11-2947
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A novel thermostable multidomain 1,4-β-xylanase from ‘Caldibacillus cellulovorans’ and effect of its xylan-binding domain on enzyme activity
Microbiology 146, 2947 (2000); https://doi.org/10.1099/00221287-146-11-2947
/content/journal/micro/10.1099/00221287-146-11-2947
/content/journal/micro/10.1099/00221287-146-11-2947
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