A novel glycoside hydrolase 43-like enzyme from Clostridium boliviensis is an endo-xylanase and a candidate for xylooligosaccharide production from different xylan substrates
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/db4197e2-2493-4548-a477-1de172c65786
- author
- Salas-Veizaga, Daniel Martin ; Rocabado-Villegas, Leonardo Roberto ; Linares-Pastén, Javier A. LU ; Eik Gudmundsdottir, Elisabet ; Hreggvidsson, Gudmundur Oli ; Álvarez-Aliaga, Maria Teresa ; Adlercreutz, Patrick LU and Nordberg Karlsson, Eva LU
- organization
- publishing date
- 2024-03-18
- 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}}, }