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Novel thermostable GH5_34 arabinoxylanase with an atypical CBM6, displays activity on oat fibre xylan for prebiotic production

Norlander, Siri LU ; Jasilionis, Andrius LU ; Ara, Zubaida Gulshan Kazi LU ; Grey, Carl LU ; Adlercreutz, Patrick LU orcid and Karlsson, Eva Nordberg LU orcid (2023) In Glycobiology 33(6). p.490-502
Abstract
Carbohydrate active enzymes are valuable tools in cereal processing to
valorise underutilized side streams. By solubilizing hemicellulose and
modifying the fibre structure, novel food products with increased
nutritional value can be created. In this study, a novel GH5_34
subfamily arabinoxylanase from Herbinix hemicellulosilytica, HhXyn5A,
was identified, produced and extensively characterized, for the
intended exploitation in cereal processing to solubilize potential
prebiotic fibres; arabinoxylo-oligosaccharides (AXOS). The purified
two-domain HhXyn5A (catalytic domain and CBM6) demonstrated high storage stability, showed a melting temperature Tm of 61 °C and... (More)
Carbohydrate active enzymes are valuable tools in cereal processing to
valorise underutilized side streams. By solubilizing hemicellulose and
modifying the fibre structure, novel food products with increased
nutritional value can be created. In this study, a novel GH5_34
subfamily arabinoxylanase from Herbinix hemicellulosilytica, HhXyn5A,
was identified, produced and extensively characterized, for the
intended exploitation in cereal processing to solubilize potential
prebiotic fibres; arabinoxylo-oligosaccharides (AXOS). The purified
two-domain HhXyn5A (catalytic domain and CBM6) demonstrated high storage stability, showed a melting temperature Tm of 61 °C and optimum reaction conditions were determined to 55 °C and pH 6.5 on wheat arabinoxylan (WAX). HhXyn5A
demonstrated activity on various commercial cereal arabinoxylans and
produced prebiotic AXOS, while the sole catalytic domain of HhXyn5A did not demonstrate detectable activity. HhXyn5A demonstrated no side activity on oat β-glucan. In contrast to the commercially available homologue CtXyn5A, HhXyn5A
gave a more specific HPAEC–PAD oligosaccharide product profile when
using WAX and alkali extracted oat bran fibres as substrate. Results
from multiple sequence alignment of GH5_34 enzymes, homology modelling
of HhXyn5A and docking simulations with ligands XXXA3, XXXA3XX, and X5, concluded that the active site of HhXyl5A
catalytic domain is highly conserved and can accommodate both shorter
and longer AXOS ligands. However, significant structural dissimilarities
between HhXyn5A and CtXyn5A in the binding cleft of
CBM6, due to lack of important ligand interacting residues, is suggested
to cause the observed differences in substrate specificity and product
formation. (Less)
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author
; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Glycobiology
volume
33
issue
6
pages
490 - 502
publisher
Oxford University Press
external identifiers
  • scopus:85164041936
  • pmid:36504389
ISSN
1460-2423
DOI
10.1093/glycob/cwac080
language
English
LU publication?
yes
additional info
© The Author(s) 2022. Published by Oxford University Press.
id
90d0d194-8705-45de-9034-ffca6870719f
date added to LUP
2023-01-23 20:46:49
date last changed
2024-06-15 06:59:38
@article{90d0d194-8705-45de-9034-ffca6870719f,
  abstract     = {{Carbohydrate active enzymes are valuable tools in cereal processing to <br>
valorise underutilized side streams. By solubilizing hemicellulose and <br>
modifying the fibre structure, novel food products with increased <br>
nutritional value can be created. In this study, a novel GH5_34 <br>
subfamily arabinoxylanase from <em>Herbinix hemicellulosilytica</em>, <em>Hh</em>Xyn5A,<br>
 was identified, produced and extensively characterized, for the <br>
intended exploitation in cereal processing to solubilize potential <br>
prebiotic fibres; arabinoxylo-oligosaccharides (AXOS). The purified <br>
two-domain <em>Hh</em>Xyn5A (catalytic domain and CBM6) demonstrated high storage stability, showed a melting temperature T<sub>m</sub> of 61 °C and optimum reaction conditions were determined to 55 °C and pH 6.5 on wheat arabinoxylan (WAX). <em>Hh</em>Xyn5A<br>
 demonstrated activity on various commercial cereal arabinoxylans and <br>
produced prebiotic AXOS, while the sole catalytic domain of <em>Hh</em>Xyn5A did not demonstrate detectable activity. <em>Hh</em>Xyn5A demonstrated no side activity on oat β-glucan. In contrast to the commercially available homologue <em>Ct</em>Xyn5A, <em>Hh</em>Xyn5A<br>
 gave a more specific HPAEC–PAD oligosaccharide product profile when <br>
using WAX and alkali extracted oat bran fibres as substrate. Results <br>
from multiple sequence alignment of GH5_34 enzymes, homology modelling <br>
of <em>Hh</em>Xyn5A and docking simulations with ligands XXXA<sup>3</sup>, XXXA<sup>3</sup>XX, and X<sup>5</sup>, concluded that the active site of <em>Hh</em>Xyl5A<br>
 catalytic domain is highly conserved and can accommodate both shorter <br>
and longer AXOS ligands. However, significant structural dissimilarities<br>
 between <em>Hh</em>Xyn5A and <em>Ct</em>Xyn5A in the binding cleft of <br>
CBM6, due to lack of important ligand interacting residues, is suggested<br>
 to cause the observed differences in substrate specificity and product <br>
formation.}},
  author       = {{Norlander, Siri and Jasilionis, Andrius and Ara, Zubaida Gulshan Kazi and Grey, Carl and Adlercreutz, Patrick and Karlsson, Eva Nordberg}},
  issn         = {{1460-2423}},
  language     = {{eng}},
  number       = {{6}},
  pages        = {{490--502}},
  publisher    = {{Oxford University Press}},
  series       = {{Glycobiology}},
  title        = {{Novel thermostable GH5_34 arabinoxylanase with an atypical CBM6, displays activity on oat fibre xylan for prebiotic production}},
  url          = {{http://dx.doi.org/10.1093/glycob/cwac080}},
  doi          = {{10.1093/glycob/cwac080}},
  volume       = {{33}},
  year         = {{2023}},
}