Teo, Seng Chong, Liew, Kok Jun, Shamsir, Mohd Shahir et al. (4 more authors) (2019) Characterizing a Halo-Tolerant GH10 Xylanase from Roseithermus sacchariphilus Strain RA and Its CBM-Truncated Variant. International Journal of Molecular Sciences. ISSN 1422-0067
Abstract
A halo-thermophilic bacterium, Roseithermus sacchariphilus strain RA (previously known as Rhodothermaceae bacterium RA), was isolated from a hot spring in Langkawi, Malaysia. A complete genome analysis showed that the bacterium harbors 57 glycoside hydrolases (GHs), including a multi-domain xylanase (XynRA2). The full-length XynRA2 of 813 amino acids comprises a family 4_9 carbohydrate-binding module (CBM4_9), a family 10 glycoside hydrolase catalytic domain (GH10), and a C-terminal domain (CTD) for type IX secretion system (T9SS). This study aims to describe the biochemical properties of XynRA2 and the effects of CBM truncation on this xylanase. XynRA2 and its CBM-truncated variant (XynRA2∆CBM) was expressed, purified, and characterized. The purified XynRA2 and XynRA2∆CBM had an identical optimum temperature at 70 ◦C, but different optimum pHs of 8.5 and 6.0 respectively. Furthermore, XynRA2 retained 94% and 71% of activity at 4.0 M and 5.0 M NaCl respectively, whereas XynRA2∆CBM showed a lower activity (79% and 54%). XynRA2 exhibited a turnover rate (kcat) of 24.8 s−1, but this was reduced by 40% for XynRA2∆CBM. Both the xylanases hydrolyzed beechwood xylan predominantly into xylobiose, and oat-spelt xylan into a mixture of xylo-oligosaccharides (XOs). Collectively, this work suggested CBM4_9 of XynRA2 has a role in enzyme performance.
Metadata
Item Type: | Article |
---|---|
Authors/Creators: |
|
Copyright, Publisher and Additional Information: | © 2019 by the authors. |
Dates: |
|
Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Biology (York) > Centre for Novel Agricultural Products (CNAP) (York) |
Funding Information: | Funder Grant number BBSRC (BIOTECHNOLOGY AND BIOLOGICAL SCIENCES RESEARCH COUNCIL) BB/P027717/1 |
Depositing User: | Pure (York) |
Date Deposited: | 17 May 2019 15:50 |
Last Modified: | 27 Dec 2024 00:12 |
Published Version: | https://doi.org/10.3390/ijms20092284 |
Status: | Published |
Refereed: | Yes |
Identification Number: | 10.3390/ijms20092284 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:146240 |
Download
Filename: Bruce_Roseithermus_xynRA2_IJMS.pdf
Description: Bruce - Roseithermus xynRA2 IJMS
Licence: CC-BY 2.5