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Characterizing a Halo-Tolerant GH10 Xylanase from Roseithermus sacchariphilus Strain RA and Its CBM-Truncated Variant
- Source :
- International Journal of Molecular Sciences, Vol 20, Iss 9, p 2284 (2019), International Journal of Molecular Sciences, Volume 20, Issue 9
- Publication Year :
- 2019
- Publisher :
- MDPI AG, 2019.
-
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&Delta<br />CBM) was expressed, purified, and characterized. The purified XynRA2 and XynRA2&Delta<br />CBM had an identical optimum temperature at 70 &deg<br />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&Delta<br />CBM showed a lower activity (79% and 54%). XynRA2 exhibited a turnover rate (kcat) of 24.8 s&minus<br />1, but this was reduced by 40% for XynRA2&Delta<br />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.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Static Electricity
01 natural sciences
Article
Catalysis
Substrate Specificity
Inorganic Chemistry
lcsh:Chemistry
03 medical and health sciences
chemistry.chemical_compound
Hydrolysis
Protein Domains
010608 biotechnology
Xylobiose
Glycoside hydrolase
glycoside hydrolase
Amino Acid Sequence
Enzyme kinetics
Physical and Theoretical Chemistry
CBM truncation
Molecular Biology
lcsh:QH301-705.5
Phylogeny
Spectroscopy
carbohydrate-binding module
xylanase
Endo-1,4-beta Xylanases
Bacteria
Strain (chemistry)
Organic Chemistry
Genetic Variation
Salt Tolerance
General Medicine
Xylan
Recombinant Proteins
Computer Science Applications
Kinetics
030104 developmental biology
chemistry
Biochemistry
lcsh:Biology (General)
lcsh:QD1-999
Xylanase
Mutant Proteins
Xylans
Carbohydrate-binding module
xylan hydrolysis
halo-tolerant
Subjects
Details
- Language :
- English
- ISSN :
- 14220067
- Volume :
- 20
- Issue :
- 9
- Database :
- OpenAIRE
- Journal :
- International Journal of Molecular Sciences
- Accession number :
- edsair.doi.dedup.....ae9abfd353502b98c9ba50af0f9a9df3