Dynamic and Functional Profiling of Xylan-Degrading Enzymes in Aspergillus Secretomes Using Activity-Based Probes

• Published in: ACS central science Vol. 5; no. 6; pp. 1067 - 1078
• Main Authors: Schröder, Sybrin P, de Boer, Casper, McGregor, Nicholas G. S, Rowland, Rhianna J, Moroz, Olga, Blagova, Elena, Reijngoud, Jos, Arentshorst, Mark, Osborn, David, Morant, Marc D, Abbate, Eric, Stringer, Mary A, Krogh, Kristian B. R. M, Raich, Lluís, Rovira, Carme, Berrin, Jean-Guy, van Wezel, Gilles P, Ram, Arthur F. J, Florea, Bogdan I, van der Marel, Gijsbert A, Codée, Jeroen D. C, Wilson, Keith S, Wu, Liang, Davies, Gideon J, Overkleeft, Herman S
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 26-06-2019; ACS Publications
• Subjects: Chemical and Process Engineering; Engineering Sciences; Food engineering; Life Sciences; enzymes; glycoside hydrolases; biomass breakdown; β-glucosidase inhibitor
• ISSN: 2374-7943; 2374-7951; 2374-7951
• DOI: 10.1021/acscentsci.9b00221

Plant polysaccharides represent a virtually unlimited feedstock for the generation of biofuels and other commodities. However, the extraordinary recalcitrance of plant polysaccharides toward breakdown necessitates a continued search for enzymes that degrade these materials efficiently under defined conditions. Activity-based protein profiling provides a route for the functional discovery of such enzymes in complex mixtures and under industrially relevant conditions. Here, we show the detection and identification of β-xylosidases and endo-β-1,4-xylanases in the secretomes of Aspergillus niger, by the use of chemical probes inspired by the β-glucosidase inhibitor cyclophellitol. Furthermore, we demonstrate the use of these activity-based probes (ABPs) to assess enzyme–substrate specificities, thermal stabilities, and other biotechnologically relevant parameters. Our experiments highlight the utility of ABPs as promising tools for the discovery of relevant enzymes useful for biomass breakdown.