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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.

Application of carbohydrate arrays coupled with mass spectrometry to detect activity of plant-polysaccharide degradative enzymes from the fungus Aspergillus niger Thumbnail


Authors

Jolanda M. van Munster

Baptiste Thomas

Michel Riese

Andrienne L. Davis

Christopher J. Gray

David B. Archer

Sabine L. Flitsch



Abstract

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.

Journal Article Type Article
Acceptance Date Jan 1, 2017
Online Publication Date Feb 21, 2017
Deposit Date Apr 26, 2017
Publicly Available Date Apr 26, 2017
Journal Scientific Reports
Electronic ISSN 2045-2322
Publisher Nature Publishing Group
Peer Reviewed Peer Reviewed
Volume 7
Issue 43117
DOI https://doi.org/10.1038/srep43117
Public URL https://nottingham-repository.worktribe.com/output/845057
Publisher URL https://www.nature.com/articles/srep43117

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