000K utf8 1100 2022$c2022-11-16 1500 eng 2050 urn:nbn:de:hbz:465-20230926-155358-5 2051 10.1038/s42003-022-04141-x 3000 Schmerling, Christian 3010 Begerow, Dominik 3010 Bräsen, Christopher 3010 Heilmann, Geronimo 3010 Jensen, Kenneth 3010 Kaiser, Markus 3010 Kaschani, Farnusch 3010 Overkleeft, Herman S. 3010 Sewald, Leonard 3010 Siebers, Bettina 3010 Witfeld, Frederick 4000 Identification of fungal lignocellulose-degrading biocatalysts secreted by Phanerochaete chrysosporium via activity-based protein profiling [Schmerling, Christian] 4209 Activity-based protein profiling (ABPP) has emerged as a versatile biochemical method for studying enzyme activity under various physiological conditions, with applications so far mainly in biomedicine. Here, we show the potential of ABPP in the discovery of biocatalysts from the thermophilic and lignocellulose-degrading white rot fungus Phanerochaete chrysosporium. By employing a comparative ABPP-based functional screen, including a direct profiling of wood substrate-bound enzymes, we identify those lignocellulose-degrading carbohydrate esterase (CE1 and CE15) and glycoside hydrolase (GH3, GH5, GH16, GH17, GH18, GH25, GH30, GH74 and GH79) enzymes specifically active in presence of the substrate. As expression of fungal enzymes remains challenging, our ABPP-mediated approach represents a preselection procedure for focusing experimental efforts on the most promising biocatalysts. Furthermore, this approach may also allow the functional annotation of domains-of-unknown functions (DUFs). The ABPP-based biocatalyst screening described here may thus allow the identification of active enzymes in a process of interest and the elucidation of novel biocatalysts that share no sequence similarity to known counterparts. 4950 https://doi.org/10.1038/s42003-022-04141-x$xR$3Volltext$534 4950 https://nbn-resolving.org/urn:nbn:de:hbz:465-20230926-155358-5$xR$3Volltext$534 4961 https://duepublico2.uni-due.de/receive/duepublico_mods_00078551 5051 540