Characterization of a family 43 β-xylosidase from the xylooligosaccharide utilizing putative probiotic Weissella sp. strain 92.
Falck, Peter LU ; Linares-Pastén, Javier LU
; Adlercreutz, Patrick
LU
and Nordberg Karlsson, Eva
LU
(2015)
In Glycobiology
26(2).
p.193-202
- Abstract
- In this work we present the first XOS degrading glycoside hydrolase from Weissella, WXyn43, a two-domain enzyme from GH43. The gene was amplified from genomic DNA of the XOS utilizing Weissella strain 92, classified under the species pair Weissella cibaria/W.confusa, and expressed in Escherichia coli. The enzyme is lacking a putative signal peptide and is, from a homology model, shown to be composed of an N-terminal 5-fold ß-propeller catalytic domain and a C-terminal ß-sandwich domain of unknown function. WXyn43 hydrolysed short (1-4)-β-D-xylooligosaccharides, with similar kcat/KM for Xylobiose (X2) and xylotriose (X3) and clearly lower efficiency in xylotetraose (X4) conversion. WXyn43 displays the highest reported kcat for conversion of... (More)
- In this work we present the first XOS degrading glycoside hydrolase from Weissella, WXyn43, a two-domain enzyme from GH43. The gene was amplified from genomic DNA of the XOS utilizing Weissella strain 92, classified under the species pair Weissella cibaria/W.confusa, and expressed in Escherichia coli. The enzyme is lacking a putative signal peptide and is, from a homology model, shown to be composed of an N-terminal 5-fold ß-propeller catalytic domain and a C-terminal ß-sandwich domain of unknown function. WXyn43 hydrolysed short (1-4)-β-D-xylooligosaccharides, with similar kcat/KM for Xylobiose (X2) and xylotriose (X3) and clearly lower efficiency in xylotetraose (X4) conversion. WXyn43 displays the highest reported kcat for conversion of X3 (900 s(-1) at 37°C) and X4 (770 s(-1)), and kcat for hydrolysis of X2 (907 s(-1)) is comparable to or greater than the highest previously reported. The purified enzyme adopted a homotetrameric state in solution, while a truncated form with isolated N-terminal catalytic domain adopted a mixture of oligomeric states and lacked detectable activity. The homology model shows that residues from both domains are involved in monomer-monomer hydrogen bonds, while the bonds creating dimer-dimer interactions only involved residues from the N-terminal domain. Docking of X2 and X3 in the active site show interactions corresponding to sub-sites -1 and +1, while presence of a third subsite is unclear, but interactions between a loop and the reducing-end xylose of X3 may be present. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/8148571
- author
- Falck, Peter
LU
; Linares-Pastén, Javier
LU
; Adlercreutz, Patrick
LU
and Nordberg Karlsson, Eva
LU
- organization
- publishing date
- 2015-10-22
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Glycobiology
- volume
- 26
- issue
- 2
- pages
- 193 - 202
- publisher
- Oxford University Press
- external identifiers
-
- pmid:26494804
- scopus:84960353803
- wos:000372013200007
- pmid:26494804
- ISSN
- 1460-2423
- DOI
- 10.1093/glycob/cwv092
- project
- ANTIDIABETIC FOOD CENTRE
- language
- English
- LU publication?
- yes
- id
- 4a7eae54-6c53-454c-87cd-03ff77a3d40a (old id 8148571)
- date added to LUP
- 2016-04-01 14:39:49
- date last changed
- 2022-04-22 04:35:14
@article{4a7eae54-6c53-454c-87cd-03ff77a3d40a,
abstract = {{In this work we present the first XOS degrading glycoside hydrolase from Weissella, WXyn43, a two-domain enzyme from GH43. The gene was amplified from genomic DNA of the XOS utilizing Weissella strain 92, classified under the species pair Weissella cibaria/W.confusa, and expressed in Escherichia coli. The enzyme is lacking a putative signal peptide and is, from a homology model, shown to be composed of an N-terminal 5-fold ß-propeller catalytic domain and a C-terminal ß-sandwich domain of unknown function. WXyn43 hydrolysed short (1-4)-β-D-xylooligosaccharides, with similar kcat/KM for Xylobiose (X2) and xylotriose (X3) and clearly lower efficiency in xylotetraose (X4) conversion. WXyn43 displays the highest reported kcat for conversion of X3 (900 s(-1) at 37°C) and X4 (770 s(-1)), and kcat for hydrolysis of X2 (907 s(-1)) is comparable to or greater than the highest previously reported. The purified enzyme adopted a homotetrameric state in solution, while a truncated form with isolated N-terminal catalytic domain adopted a mixture of oligomeric states and lacked detectable activity. The homology model shows that residues from both domains are involved in monomer-monomer hydrogen bonds, while the bonds creating dimer-dimer interactions only involved residues from the N-terminal domain. Docking of X2 and X3 in the active site show interactions corresponding to sub-sites -1 and +1, while presence of a third subsite is unclear, but interactions between a loop and the reducing-end xylose of X3 may be present.}},
author = {{Falck, Peter and Linares-Pastén, Javier and Adlercreutz, Patrick and Nordberg Karlsson, Eva}},
issn = {{1460-2423}},
language = {{eng}},
month = {{10}},
number = {{2}},
pages = {{193--202}},
publisher = {{Oxford University Press}},
series = {{Glycobiology}},
title = {{Characterization of a family 43 β-xylosidase from the xylooligosaccharide utilizing putative probiotic Weissella sp. strain 92.}},
url = {{http://dx.doi.org/10.1093/glycob/cwv092}},
doi = {{10.1093/glycob/cwv092}},
volume = {{26}},
year = {{2015}},
}