Narin Kirikyali
Characterisation of a recombinant β-xylosidase (xylA) from Aspergillus oryzae expressed in Pichia pastoris
Kirikyali, Narin; Wood, Jonathan; Connerton, Ian F.
Authors
Abstract
β-xylosidases catalyse the hydrolysis of short chain xylooligosaccharides from their non-reducing ends into xylose. In this study we report the heterologous expression of Aspergillus oryzae β-xylosidase (XylA) in Pichia pastoris under the control of the glyceraldehyde-3-phosphate dehydrogenase promoter. The recombinant enzyme was optimally active at 55°C and pH 4.5 with Km and Vmax values of 1.0 mM and 250 μmol min−1 mg−1 respectively against 4-nitrophenyl β-xylopyranoside. Xylose was a competitive inhibitor with a Ki of 2.72 mM, whereas fructose was an uncompetitive inhibitor reducing substrate binding affinity (Km) and conversion efficiency (Vmax). The enzyme was characterised to be an exo-cutting enzyme releasing xylose from the non-reducing ends of β-1,4 linked xylooligosaccharides (X2, X3 and X4). Catalytic conversion of X2, X3 and X4 decreased (Vmax and kcat) with increasing chain length.
Citation
Kirikyali, N., Wood, J., & Connerton, I. F. (2014). Characterisation of a recombinant β-xylosidase (xylA) from Aspergillus oryzae expressed in Pichia pastoris. AMB Express, 4(68), https://doi.org/10.1186/s13568-014-0068-1
Journal Article Type | Article |
---|---|
Publication Date | Aug 31, 2014 |
Deposit Date | May 22, 2015 |
Publicly Available Date | May 22, 2015 |
Journal | AMB Express |
Electronic ISSN | 2191-0855 |
Publisher | Springer Verlag |
Peer Reviewed | Peer Reviewed |
Volume | 4 |
Issue | 68 |
DOI | https://doi.org/10.1186/s13568-014-0068-1 |
Public URL | https://nottingham-repository.worktribe.com/output/733589 |
Publisher URL | http://www.amb-express.com/content/4/1/68 |
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Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0
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