RT Journal Article SR Electronic(1) A1 Sunna, Anwar A1 Gibbs, Moreland D. A1 Bergquist, Peter L. YR 2000 T1 A novel thermostable multidomain 1,4-β-xylanase from ‘Caldibacillus cellulovorans’ and effect of its xylan-binding domain on enzyme activity JF Microbiology, VO 146 IS 11 SP 2947 OP 2955 DO https://doi.org/10.1099/00221287-146-11-2947 PB Microbiology Society, SN 1465-2080, AB The GenBank accession number for the sequence reported in this paper is AF200304. The nucleotide sequence of the complete xynA gene, encoding a novel multidomain xylanase XynA of ‘Caldibacillus cellulovorans’, 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 xynA 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., UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-146-11-2947