Carbohydrate arrays coupled with mass spectrometry detect activity of plant polysaccharide degradative enzymes of Aspergillus niger

van Munster, Jolanda M. and Archer, David B. (2017) Carbohydrate arrays coupled with mass spectrometry detect activity of plant polysaccharide degradative enzymes of Aspergillus niger. Scientific Reports, 7 (43117). pp. 1-13. ISSN 2045-2322

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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.

Item Type: Article
Schools/Departments: University of Nottingham, UK > Faculty of Science > School of Chemistry
University of Nottingham, UK > Faculty of Medicine and Health Sciences > School of Life Sciences
Identification Number: 10.1038/srep43117
Depositing User: Archer, David
Date Deposited: 26 Apr 2017 10:52
Last Modified: 13 Oct 2017 00:59
URI: http://eprints.nottingham.ac.uk/id/eprint/42228

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