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Novel Function of CtXyn5A from Acetivibrio thermocellus : Dual Arabinoxylanase and Feruloyl Esterase Activity in the Same Active Site

Schmitz, Eva LU ; Leontakianakou, Savvina LU ; Adlercreutz, Patrick LU orcid ; Nordberg Karlsson, Eva LU orcid and Linares-Pastén, Javier LU orcid (2023) In ChemBioChem 24(3).
Abstract

Uncharacterized side activities of enzymes can have significant effects on reaction products and yields. Hence, their identification and characterization are crucial for the development of successful reaction systems. Here, we report the presence of feruloyl esterase activity in CtXyn5A from Acetivibrio thermocellus, besides its well-known arabinoxylanase activity, for the first time. Activity analysis of enzyme variants mutated in the catalytic nucleophile, Glu279, confirmed removal of all activity for E279A and E279L, and increased esterase activity while removing xylanase activity for E279S, allowing the proposal that both reaction types are catalysed in the same active site in two subsequential steps. The ferulic acid substituent is... (More)

Uncharacterized side activities of enzymes can have significant effects on reaction products and yields. Hence, their identification and characterization are crucial for the development of successful reaction systems. Here, we report the presence of feruloyl esterase activity in CtXyn5A from Acetivibrio thermocellus, besides its well-known arabinoxylanase activity, for the first time. Activity analysis of enzyme variants mutated in the catalytic nucleophile, Glu279, confirmed removal of all activity for E279A and E279L, and increased esterase activity while removing xylanase activity for E279S, allowing the proposal that both reaction types are catalysed in the same active site in two subsequential steps. The ferulic acid substituent is cleaved off first, followed by hydrolysis of the xylan backbone. The esterase activity on complex carbohydrates was found to be higher than the one of a designated ferulic acid esterase (E-FAERU). Therefore, we conclude that the enzyme exhibits a dual function rather than an esterase side activity.

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author
; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
ChemBioChem
volume
24
issue
3
publisher
John Wiley & Sons Inc.
external identifiers
  • pmid:36449982
  • scopus:85145075978
ISSN
1439-4227
DOI
10.1002/cbic.202200667
language
English
LU publication?
yes
additional info
© 2022 Wiley-VCH GmbH.
id
69694a27-948f-46c1-9824-cea2fcea0078
date added to LUP
2022-12-15 13:43:14
date last changed
2024-06-12 17:02:42
@article{69694a27-948f-46c1-9824-cea2fcea0078,
  abstract     = {{<p>Uncharacterized side activities of enzymes can have significant effects on reaction products and yields. Hence, their identification and characterization are crucial for the development of successful reaction systems. Here, we report the presence of feruloyl esterase activity in CtXyn5A from Acetivibrio thermocellus, besides its well-known arabinoxylanase activity, for the first time. Activity analysis of enzyme variants mutated in the catalytic nucleophile, Glu279, confirmed removal of all activity for E279A and E279L, and increased esterase activity while removing xylanase activity for E279S, allowing the proposal that both reaction types are catalysed in the same active site in two subsequential steps. The ferulic acid substituent is cleaved off first, followed by hydrolysis of the xylan backbone. The esterase activity on complex carbohydrates was found to be higher than the one of a designated ferulic acid esterase (E-FAERU). Therefore, we conclude that the enzyme exhibits a dual function rather than an esterase side activity.</p>}},
  author       = {{Schmitz, Eva and Leontakianakou, Savvina and Adlercreutz, Patrick and Nordberg Karlsson, Eva and Linares-Pastén, Javier}},
  issn         = {{1439-4227}},
  language     = {{eng}},
  number       = {{3}},
  publisher    = {{John Wiley & Sons Inc.}},
  series       = {{ChemBioChem}},
  title        = {{Novel Function of <i>Ct</i>Xyn5A from <i>Acetivibrio thermocellus</i> : Dual Arabinoxylanase and Feruloyl Esterase Activity in the Same Active Site}},
  url          = {{http://dx.doi.org/10.1002/cbic.202200667}},
  doi          = {{10.1002/cbic.202200667}},
  volume       = {{24}},
  year         = {{2023}},
}