Characterisation of a recombinant β-xylosidase (xylA) from Aspergillus oryzae expressed in Pichia pastoris
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 promo...
| Main Authors: | , , |
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| Format: | Article |
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SpringerOpen
2014
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| Online Access: | https://eprints.nottingham.ac.uk/28794/ |
| _version_ | 1848793643427561472 |
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| author | Kirikyali, Narin Wood, Jonathan Connerton, Ian F. |
| author_facet | Kirikyali, Narin Wood, Jonathan Connerton, Ian F. |
| author_sort | Kirikyali, Narin |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T19:03:34Z |
| format | Article |
| id | nottingham-28794 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:03:34Z |
| publishDate | 2014 |
| publisher | SpringerOpen |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-287942020-05-04T16:51:55Z https://eprints.nottingham.ac.uk/28794/ Characterisation of a recombinant β-xylosidase (xylA) from Aspergillus oryzae expressed in Pichia pastoris Kirikyali, Narin Wood, Jonathan Connerton, Ian F. 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. SpringerOpen 2014-08-31 Article PeerReviewed Kirikyali, Narin, Wood, Jonathan and Connerton, Ian F. (2014) Characterisation of a recombinant β-xylosidase (xylA) from Aspergillus oryzae expressed in Pichia pastoris. AMB Express, 4 (68). pp. 1-7. ISSN 2191-0855 http://www.amb-express.com/content/4/1/68 doi:10.1186/s13568-014-0068-1 doi:10.1186/s13568-014-0068-1 |
| spellingShingle | Kirikyali, Narin Wood, Jonathan Connerton, Ian F. Characterisation of a recombinant β-xylosidase (xylA) from Aspergillus oryzae expressed in Pichia pastoris |
| title | Characterisation of a recombinant β-xylosidase
(xylA) from Aspergillus oryzae expressed in Pichia
pastoris |
| title_full | Characterisation of a recombinant β-xylosidase
(xylA) from Aspergillus oryzae expressed in Pichia
pastoris |
| title_fullStr | Characterisation of a recombinant β-xylosidase
(xylA) from Aspergillus oryzae expressed in Pichia
pastoris |
| title_full_unstemmed | Characterisation of a recombinant β-xylosidase
(xylA) from Aspergillus oryzae expressed in Pichia
pastoris |
| title_short | Characterisation of a recombinant β-xylosidase
(xylA) from Aspergillus oryzae expressed in Pichia
pastoris |
| title_sort | characterisation of a recombinant β-xylosidase
(xyla) from aspergillus oryzae expressed in pichia
pastoris |
| url | https://eprints.nottingham.ac.uk/28794/ https://eprints.nottingham.ac.uk/28794/ https://eprints.nottingham.ac.uk/28794/ |