Application of carbohydrate arrays coupled with mass spectrometry to detect activity of plant-polysaccharide degradative enzymes from the fungus Aspergillus niger
Renewables-based biotechnology depends on enzymes to degrade plant lignocellulose to simple sugars that are converted to fuels or high-value products. Identification and characterization of such lignocellulose degradative enzymes could be fast-tracked by availability of an enzyme activity measuremen...
| Main Authors: | , , , , , , |
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| Format: | Article |
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Nature Publishing Group
2017
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| Online Access: | https://eprints.nottingham.ac.uk/42228/ |
| _version_ | 1848796448724877312 |
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| author | van Munster, Jolanda M. Thomas, Baptiste Riese, Michel Davis, Andrienne L. Gray, Christopher J. Archer, David B. Flitsch, Sabine L. |
| author_facet | van Munster, Jolanda M. Thomas, Baptiste Riese, Michel Davis, Andrienne L. Gray, Christopher J. Archer, David B. Flitsch, Sabine L. |
| author_sort | van Munster, Jolanda M. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Renewables-based biotechnology depends on enzymes to degrade plant lignocellulose to simple sugars that are converted to fuels or high-value products. Identification and characterization of such lignocellulose degradative enzymes could be fast-tracked by availability of an enzyme activity measurement method that is fast, label-free, uses minimal resources and allows direct identificationof generated products. We developed such a method by applying carbohydrate arrays coupled with MALDI-ToF mass spectrometry to identify reaction products of carbohydrate active enzymes (CAZymes) of the filamentous fungus Aspergillus niger. We describe the production and characterization of plant polysaccharide-derived oligosaccharides and their attachment to hydrophobic self-assembling monolayers on a gold target. We verify effectiveness of this array for detecting exo- and endo-acting glycoside hydrolase activity using commercial enzymes, and demonstrate how this platform is suitable for detection of enzyme activity in relevant biological samples, the culture filtrate of A. niger grown on wheat straw. In conclusion, this versatile method is broadly applicable in screening and characterisation of activity of CAZymes, such as fungal enzymes for plant lignocellulose degradation with relevance to biotechnological applications as biofuel production, the food and animal feed industry. |
| first_indexed | 2025-11-14T19:48:09Z |
| format | Article |
| id | nottingham-42228 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:48:09Z |
| publishDate | 2017 |
| publisher | Nature Publishing Group |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-422282020-05-04T18:34:07Z https://eprints.nottingham.ac.uk/42228/ Application of carbohydrate arrays coupled with mass spectrometry to detect activity of plant-polysaccharide degradative enzymes from the fungus Aspergillus niger van Munster, Jolanda M. Thomas, Baptiste Riese, Michel Davis, Andrienne L. Gray, Christopher J. Archer, David B. Flitsch, Sabine L. Renewables-based biotechnology depends on enzymes to degrade plant lignocellulose to simple sugars that are converted to fuels or high-value products. Identification and characterization of such lignocellulose degradative enzymes could be fast-tracked by availability of an enzyme activity measurement method that is fast, label-free, uses minimal resources and allows direct identificationof generated products. We developed such a method by applying carbohydrate arrays coupled with MALDI-ToF mass spectrometry to identify reaction products of carbohydrate active enzymes (CAZymes) of the filamentous fungus Aspergillus niger. We describe the production and characterization of plant polysaccharide-derived oligosaccharides and their attachment to hydrophobic self-assembling monolayers on a gold target. We verify effectiveness of this array for detecting exo- and endo-acting glycoside hydrolase activity using commercial enzymes, and demonstrate how this platform is suitable for detection of enzyme activity in relevant biological samples, the culture filtrate of A. niger grown on wheat straw. In conclusion, this versatile method is broadly applicable in screening and characterisation of activity of CAZymes, such as fungal enzymes for plant lignocellulose degradation with relevance to biotechnological applications as biofuel production, the food and animal feed industry. Nature Publishing Group 2017-02-21 Article PeerReviewed van Munster, Jolanda M., Thomas, Baptiste, Riese, Michel, Davis, Andrienne L., Gray, Christopher J., Archer, David B. and Flitsch, Sabine L. (2017) Application of carbohydrate arrays coupled with mass spectrometry to detect activity of plant-polysaccharide degradative enzymes from the fungus Aspergillus niger. Scientific Reports, 7 (43117). pp. 1-13. ISSN 2045-2322 https://www.nature.com/articles/srep43117 doi:10.1038/srep43117 doi:10.1038/srep43117 |
| spellingShingle | van Munster, Jolanda M. Thomas, Baptiste Riese, Michel Davis, Andrienne L. Gray, Christopher J. Archer, David B. Flitsch, Sabine L. Application of carbohydrate arrays coupled with mass spectrometry to detect activity of plant-polysaccharide degradative enzymes from the fungus Aspergillus niger |
| title | Application of carbohydrate arrays coupled with mass spectrometry to detect activity of plant-polysaccharide degradative enzymes from the fungus Aspergillus niger |
| title_full | Application of carbohydrate arrays coupled with mass spectrometry to detect activity of plant-polysaccharide degradative enzymes from the fungus Aspergillus niger |
| title_fullStr | Application of carbohydrate arrays coupled with mass spectrometry to detect activity of plant-polysaccharide degradative enzymes from the fungus Aspergillus niger |
| title_full_unstemmed | Application of carbohydrate arrays coupled with mass spectrometry to detect activity of plant-polysaccharide degradative enzymes from the fungus Aspergillus niger |
| title_short | Application of carbohydrate arrays coupled with mass spectrometry to detect activity of plant-polysaccharide degradative enzymes from the fungus Aspergillus niger |
| title_sort | application of carbohydrate arrays coupled with mass spectrometry to detect activity of plant-polysaccharide degradative enzymes from the fungus aspergillus niger |
| url | https://eprints.nottingham.ac.uk/42228/ https://eprints.nottingham.ac.uk/42228/ https://eprints.nottingham.ac.uk/42228/ |