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Dynamic and functional profiling of xylan-degrading enzymes in Aspergillus secretomes using activity-based probes

DOI: 10.1021/acscentsci.9b00221 DOI Help

Authors: Sybrin P. Schröder (Leiden University) , Casper De Boer (Leiden University) , Nicholas G. S. Mcgregor (The University of York) , Rhianna J. Rowland (The University of York) , Olga Moroz (The University of York) , Elena Blagova (The University of York) , Jos Reijngoud (Leiden University) , Mark Arentshorst (Leiden University) , David Osborn (Novozymes Inc) , Marc D. Morant (Novozymes A/S) , Eric Abbate (Novozymes Inc) , Mary A. Stringer (Novozymes Inc) , Kristian B. R. M. Krogh (Novozymes Inc) , Lluís Raich (Universitat de Barcelona) , Carme Rovira (Universitat de Barcelona; Institució Catalana de Recerca i Estudis Avançats (ICREA)) , Jean-guy Berrin (Aix Marseille University) , Gilles P. Van Wezel (Leiden University) , Arthur F. J. Ram (Leiden University) , Bogdan I. Florea (Leiden University) , Gijsbert A. Van Der Marel (Leiden University) , Jeroen D. C. Codée (Leiden University) , Keith S. Wilson (The University of York) , Liang Wu (The University of York) , Gideon J. Davies (The University of York) , Herman S. Overkleeft (Leiden University)
Co-authored by industrial partner: Yes

Type: Journal Paper
Journal: Acs Central Science

State: Published (Approved)
Published: May 2019
Diamond Proposal Number(s): 13587

Abstract: 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.

Subject Areas: Biology and Bio-materials, Chemistry, Energy

Instruments: I03-Macromolecular Crystallography , I04-Macromolecular Crystallography