Plant polysaccharides represent a virtually unlimited feedstock for the generation of biofuels and other commodities. However, the extraordinary recalcitrance of plant polysaccharides towards breakdown necessitates a continued search for enzymes that degrade these materials efficiently under defined conditions. Activity-based protein profiling (ABPP) 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 b-xylosidases and endo-xylanases in the secretome of Aspergillus niger, by the use of chemical probes inspired by the b-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.
[doi:10.25345/C5ZS7S]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: Activity-based probe ; Glycosidase ; Biofuel ; Industrial enzyme ; Xylanase
Principal Investigators: (in alphabetical order) |
Gideon J. Davies, University of York, United Kingdom |
Submitting User: | aad100 |
Schröder SP, de Boer C, McGregor NGS, Rowland RJ, Moroz O, Blagova E, Reijngoud J, Arentshorst M, Osborn D, Morant MD, Abbate E, Stringer MA, Krogh KBRM, Raich L, Rovira C, Berrin JG, van Wezel GP, Ram AFJ, Florea BI, van der Marel GA, Codée JDC, Wilson KS, Wu L, Davies GJ, Overkleeft HS.
Dynamic and Functional Profiling of Xylan-Degrading Enzymes in Aspergillus Secretomes Using Activity-Based Probes.
ACS Cent Sci. 2019 Jun 26;5(6):1067-1078. Epub 2019 May 24.
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