A novel thermostable multidomain 1,4-β-xylanase from 'Caldibacillus cellulovorans' and effect of its xylan-binding domain on enzyme activity

A. Sunna, M. D. Gibbs, P. L. Bergquist

Research output: Contribution to journalArticleResearchpeer-review

Abstract

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 (XynAd 1/2 ) 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 (XynAd 1/2 ) 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.

LanguageEnglish
Pages2947-2955
Number of pages9
JournalMicrobiology
Volume146
Issue number11
Publication statusPublished - 2000

Fingerprint

Endo-1,4-beta Xylanases
Xylans
Catalytic Domain
Enzymes
Cellulose
Hydrolases
Threonine
Substrate Specificity
Proline
Open Reading Frames
Walking
Hydrolysis
Plasmids
Polymerase Chain Reaction
Peptides
Genes

Cite this

@article{ca0f12e3b0014a66a5f468f57d0090b9,
title = "A novel thermostable multidomain 1,4-β-xylanase from 'Caldibacillus cellulovorans' and effect of its xylan-binding domain on enzyme activity",
abstract = "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 (XynAd 1/2 ) 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 (XynAd 1/2 ) 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.",
author = "A. Sunna and Gibbs, {M. D.} and Bergquist, {P. L.}",
year = "2000",
language = "English",
volume = "146",
pages = "2947--2955",
journal = "Microbiology",
issn = "1350-0872",
publisher = "Microbiology Society",
number = "11",

}

A novel thermostable multidomain 1,4-β-xylanase from 'Caldibacillus cellulovorans' and effect of its xylan-binding domain on enzyme activity. / Sunna, A.; Gibbs, M. D.; Bergquist, P. L.

In: Microbiology, Vol. 146, No. 11, 2000, p. 2947-2955.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - A novel thermostable multidomain 1,4-β-xylanase from 'Caldibacillus cellulovorans' and effect of its xylan-binding domain on enzyme activity

AU - Sunna, A.

AU - Gibbs, M. D.

AU - Bergquist, P. L.

PY - 2000

Y1 - 2000

N2 - 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 (XynAd 1/2 ) 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 (XynAd 1/2 ) 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.

AB - 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 (XynAd 1/2 ) 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 (XynAd 1/2 ) 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.

UR - http://www.scopus.com/inward/record.url?scp=0033677793&partnerID=8YFLogxK

M3 - Article

VL - 146

SP - 2947

EP - 2955

JO - Microbiology

T2 - Microbiology

JF - Microbiology

SN - 1350-0872

IS - 11

ER -