Moroz, Olga V., Jensen, Pernille F., McDonald, Sean P. et al. (17 more authors) (2018) Structural dynamics and catalytic properties of a multimodular xanthanase. ACS Catalysis. 6021–6034. ISSN 2155-5435
Abstract
The precise catalytic strategies used for the breakdown of the complex bacterial polysaccharide xanthan, an increasingly frequent component of processed human foodstuffs, have remained a mystery. Here, we present characterization of an endo-xanthanase from Paenibacillus nanensis. We show that it is a CAZy family 9 glycoside hydrolase (GH9) responsible for the hydrolysis of the xanthan backbone capable of generating tetrameric xanthan oligosaccharides from polysaccharide lyase family 8 (PL8) xanthan lyase-treated xanthan. Three-dimensional structure determination reveals a complex multimodular enzyme in which a catalytic (α/α) 6 barrel is flanked by an N-terminal "immunoglobulin-like" (Ig-like) domain (frequently found in GH9 enzymes) and by four additional C-terminal all β-sheet domains that have very few homologues in sequence databases and at least one of which functions as a new xanthan-binding domain, now termed CBM84. The solution-phase conformation and dynamics of the enzyme in the native calcium-bound state and in the absence of calcium were probed experimentally by hydrogen/deuterium exchange mass spectrometry. Measured conformational dynamics were used to guide the protein engineering of enzyme variants with increased stability in the absence of calcium; a property of interest for the potential use of the enzyme in cleaning detergents. The ability of hydrogen/deuterium exchange mass spectrometry to pinpoint dynamic regions of a protein under stress (e.g., removal of calcium ions) makes this technology a strong tool for improving protein catalyst properties by informed engineering.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2018 American Chemical Society. This is an author-produced version of the published paper. Uploaded in accordance with the publisher’s self-archiving policy. Further copying may not be permitted; contact the publisher for details |
Keywords: | carbohydrate,enzyme,enzyme dynamics,enzyme stability,hydrogen/deuterium exchange mass spectrometry,xanthan |
Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Chemistry (York) |
Depositing User: | Pure (York) |
Date Deposited: | 22 Jun 2018 10:40 |
Last Modified: | 28 Oct 2024 00:59 |
Published Version: | https://doi.org/10.1021/acscatal.8b00666 |
Status: | Published |
Refereed: | Yes |
Identification Number: | 10.1021/acscatal.8b00666 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:132388 |
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