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A novel thermostable GH5 β-xylosidase from Thermogemmatispora sp. T81
- Source :
- Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual), Universidade de São Paulo (USP), instacron:USP
- Publication Year :
- 2019
- Publisher :
- Elsevier BV, 2019.
-
Abstract
- A glycoside hydrolase family 5 (GH5) subfamily 22 gene, designated T81Xyl5_22A, was identified in the genome of the aerobic thermophilic bacterium, Thermogemmatispora sp. T81 (locus A4R35_07040). The gene was cloned and heterologously expressed in Escherichia coli and the gene product characterized biochemically. The recombinant enzyme had an optimal catalytic activity at pH5.0 and 65 °C, and was active against beechwood xylan and rye arabinoxylan. It yielded only xylose molecules as products of beechwood xylan hydrolysis, indicating that it is a GH5 family β- d -xylosidase. Using 4-nitrophenyl β- d -xylopyranoside (pNPX) as a substrate, the KM, Vmax, kcat and kcat/KM kinetic parameters were determined as 0.25 ± 0.03 mM, 889.47 ± 28.54 U/mg, 39.20 s−1 and 156.8 mM−1 s−1, respectively. Small-angle X-ray scattering (SAXS) data enabled reconstruction of the enzyme’s low-resolution molecular envelope and revealed that it formed dimers in solution. As far as we are aware, this is the first description of a thermostable bacterial GH5 family β- d -xylosidase.
- Subjects :
- 0106 biological sciences
Stereochemistry
Bioengineering
Xylose
medicine.disease_cause
01 natural sciences
03 medical and health sciences
chemistry.chemical_compound
010608 biotechnology
Enzyme Stability
Arabinoxylan
medicine
Enzyme kinetics
Molecular Biology
Escherichia coli
030304 developmental biology
Thermostability
chemistry.chemical_classification
0303 health sciences
Glycoside hydrolase family 5
Temperature
EXPRESSÃO GÊNICA
Chloroflexi
General Medicine
Hydrogen-Ion Concentration
Enzyme structure
Xylosidases
Enzyme
chemistry
Biotechnology
Subjects
Details
- ISSN :
- 18716784
- Volume :
- 53
- Database :
- OpenAIRE
- Journal :
- New Biotechnology
- Accession number :
- edsair.doi.dedup.....fe91fa7485c72ca393fa2cc1e004a5f7