Activity-Based Protein Profiling of Glycosidases In Extremophilic Archaea

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1st EuroCarb Young Researcher (ECYR) Workshop

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Mini symposium: Glycobiology at the LUMC and beyond

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Activity-Based Protein Profiling of Glycosidases In Extremophilic Archaea

Sabrina Ninck 1 , Thomas Klaus 1 , Farnusch Kaschani 1 , Herman Overkleeft 2 , Bettina Siebers 1 , Markus Kaiser 1

1 University of Duisburg-Essen, Essen, Germany, 2 Leiden University, Leiden, The Netherlands

Archaea build up one of the domains of life, alongside Eukarya and Bacteria[1]. They are characterised by a unique life style in often environmental extremes regarding temperature, pH, salinity or combinations thereof. To be able to colonise these habitats, Archaea have adapted their protein repertoire in a long evolutionary process. Enzymes from thermophilic Archaea are therefore for example very stable and active even under extreme temperatures. These unique properties turn archaeal ”extremozymes” into an exclusive branch of biocatalytic biodiversity, harbouring great potential for (white) biotechnology[2-3], metabolic engineering as well as synthetic biology[4].

Though their extreme lifestyle turns Archaea into an interesting field of research, they are currently underexplored and further investigations to overcome this limitation are required. The characterisation of novel archaeal glycosidases is of great interest, as these enzymes are valuable for diverse biotechnological applications such as lignocellulosic biomass degradation[5]. To promote the functional identification of potential archaeal biocatalysts involved in the degradation of carbohydrates, especially of complex polysaccharides like xylan, chemical proteomics can be used by combining activity-based protein profiling (ABPP) with mass spectrometry. The use of bioactive chemical probes allows for the labelling and purification of target enzymes. Afterwards, these target enzymes can be identified using mass spectrometry-based proteomics[6].

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Sarmiento, F.; Peralta, R.; Blamey, J. M., Cold and Hot Extremozymes: Industrial Relevance and Current Trends. Front Bioeng Biotechnol 2015, 3, 148.

Bräsen, C.; Esser, D.; Rauch, B.; Siebers, B., Carbohydrate Metabolism in Archaea: Current Insights into Unusual Enzymes and Pathways and Their Regulation. Microbiol Mol Biol R 2014, 78 (1), 89-175.

Sweeney, M. D.; Xu, F., Biomass Converting Enzymes as Industrial Biocatalysts for Fuels and Chemicals: Recent Developments. Catalysts 2012, 2 (2), 244-263.

Zweerink, S.; Kallnik, V.; Ninck, S.; Nickel, S.; Verheyen, J.; Blum, M.; Wagner, A.; Feldmann, I.; Sickmann, A.; Albers, S. V.; Brasen, C.; Kaschani, F.; Siebers, B.; Kaiser, M., Activity-based protein profiling as a robust method for enzyme identification and screening in extremophilic Archaea. Nat Commun 2017, 8.

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