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A novel glycoside hydrolase 43-like enzyme from Clostridium boliviensis is an endo-xylanase and a candidate for xylooligosaccharide production from different xylan substrates

Salas-Veizaga, Daniel Martin ; Rocabado-Villegas, Leonardo Roberto ; Linares-Pastén, Javier A. LU orcid ; Eik Gudmundsdottir, Elisabet ; Hreggvidsson, Gudmundur Oli ; Álvarez-Aliaga, Maria Teresa ; Adlercreutz, Patrick LU orcid and Nordberg Karlsson, Eva LU orcid (2024) In Applied and Environmental Microbiology
Abstract
An uncharacterized gene encoding a glycoside hydrolase family 43-like enzyme from Clostridium boliviensis strain E-1 was identified from genomic sequence data, and the encoded enzyme, CbE1Xyn43-l, was produced in Escherichia coli. CbE1Xyn43-l
(52.9 kDa) is a two-domain endo-β-xylanase consisting of a C-terminal
CBM6 and a GH43-like catalytic domain. The positions of the catalytic
dyad conserved in GH43, the catalytic base (Asp74), and proton donor
(Glu240) were identified in alignments including GH43-enzymes of known
3D-structure from different subfamilies. CbE1Xyn43-l is active at
pH 7.0–9.0, with optimum temperature at 65°C, and a more than 7 days’
half-life in irreversible... (More)
An uncharacterized gene encoding a glycoside hydrolase family 43-like enzyme from Clostridium boliviensis strain E-1 was identified from genomic sequence data, and the encoded enzyme, CbE1Xyn43-l, was produced in Escherichia coli. CbE1Xyn43-l
(52.9 kDa) is a two-domain endo-β-xylanase consisting of a C-terminal
CBM6 and a GH43-like catalytic domain. The positions of the catalytic
dyad conserved in GH43, the catalytic base (Asp74), and proton donor
(Glu240) were identified in alignments including GH43-enzymes of known
3D-structure from different subfamilies. CbE1Xyn43-l is active at
pH 7.0–9.0, with optimum temperature at 65°C, and a more than 7 days’
half-life in irreversible deactivation studies at this temperature. The
enzyme hydrolyzed birchwood xylan, quinoa stalks glucuronoarabinoxylan,
and wheat arabinoxylan with xylotriose and xylotetraose as major
hydrolysis products. CbE1Xyn43-l also released xylobiose from pNPX2 with low turnover (kcat of 0.044 s−1) but was inactive on pNPX,
showing that a degree of polymerization of three (DP3) was the smallest
hydrolyzable substrate. Divalent ions affected the specific activity on
xylan substrates, which dependent on the ion could be increased or
decreased. In conclusion, CbE1Xyn43-l from C. boliviensis
strain E-1 is the first characterized member of a large group of
homologous hypothetical proteins annotated as GH43-like and is a
thermostable endo-xylanase, producing xylooligosaccharides of high DP
(xylotriose and xylotetraose) producer. (Less)
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author
; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
epub
subject
in
Applied and Environmental Microbiology
article number
e02223-23
publisher
American Society for Microbiology
external identifiers
  • pmid:38497645
ISSN
0099-2240
DOI
10.1128/aem.02223-23
language
English
LU publication?
yes
id
db4197e2-2493-4548-a477-1de172c65786
date added to LUP
2024-03-20 17:32:00
date last changed
2024-03-22 11:41:27
@article{db4197e2-2493-4548-a477-1de172c65786,
  abstract     = {{An uncharacterized gene encoding a glycoside hydrolase family 43-like enzyme from <i>Clostridium boliviensis</i> strain E-1 was identified from genomic sequence data, and the encoded enzyme, <i>CbE1</i>Xyn43-l, was produced in <i>Escherichia coli. CbE1</i>Xyn43-l<br>
 (52.9 kDa) is a two-domain endo-β-xylanase consisting of a C-terminal <br>
CBM6 and a GH43-like catalytic domain. The positions of the catalytic <br>
dyad conserved in GH43, the catalytic base (Asp74), and proton donor <br>
(Glu240) were identified in alignments including GH43-enzymes of known <br>
3D-structure from different subfamilies. <i>CbE1</i>Xyn43-l is active at<br>
 pH 7.0–9.0, with optimum temperature at 65°C, and a more than 7 days’ <br>
half-life in irreversible deactivation studies at this temperature. The <br>
enzyme hydrolyzed birchwood xylan, quinoa stalks glucuronoarabinoxylan, <br>
and wheat arabinoxylan with xylotriose and xylotetraose as major <br>
hydrolysis products. <i>CbE1</i>Xyn43-l also released xylobiose from <i>p</i>NPX<sub>2</sub> with low turnover (<i>k</i><sub>cat</sub> of 0.044 s<sup>−1</sup>) but was inactive on <i>p</i>NPX,<br>
 showing that a degree of polymerization of three (DP3) was the smallest<br>
 hydrolyzable substrate. Divalent ions affected the specific activity on<br>
 xylan substrates, which dependent on the ion could be increased or <br>
decreased. In conclusion, <i>CbE1</i>Xyn43-l from <i>C. boliviensis</i> <br>
strain E-1 is the first characterized member of a large group of <br>
homologous hypothetical proteins annotated as GH43-like and is a <br>
thermostable endo-xylanase, producing xylooligosaccharides of high DP <br>
(xylotriose and xylotetraose) producer.}},
  author       = {{Salas-Veizaga, Daniel Martin and Rocabado-Villegas, Leonardo Roberto and Linares-Pastén, Javier A. and Eik Gudmundsdottir, Elisabet and Hreggvidsson, Gudmundur Oli and Álvarez-Aliaga, Maria Teresa and Adlercreutz, Patrick and Nordberg Karlsson, Eva}},
  issn         = {{0099-2240}},
  language     = {{eng}},
  month        = {{03}},
  publisher    = {{American Society for Microbiology}},
  series       = {{Applied and Environmental Microbiology}},
  title        = {{A novel glycoside hydrolase 43-like enzyme from <i>Clostridium boliviensis </i>is an endo-xylanase and a candidate for xylooligosaccharide production from different xylan substrates}},
  url          = {{http://dx.doi.org/10.1128/aem.02223-23}},
  doi          = {{10.1128/aem.02223-23}},
  year         = {{2024}},
}