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.

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.

Citation

van Munster, J. M., Thomas, B., Riese, M., Davis, A. L., Gray, C. J., Archer, D. B., & Flitsch, S. L. (in press). 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), https://doi.org/10.1038/srep43117

• 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
• Contract Date: Apr 26, 2017