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Three-dimensional structures of two heavily N-glycosylated Aspergillus sp family GH3 beta-D-glucosidases
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SYSNO ASEP 0470039 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Three-dimensional structures of two heavily N-glycosylated Aspergillus sp family GH3 beta-D-glucosidases Author(s) Agirre, J. (GB)
Ariza, A. (GB)
Offen, W. A. (GB)
Turkenburg, J. P. (GB)
Roberts, S. M. (GB)
McNicholas, S. (GB)
Harris, P. V. (US)
McBrayer, B. (US)
Dohnálek, Jan (BTO-N) RID, ORCID
Cowtan, K. D. (GB)
Davies, G. J. (GB)
Wilson, K. S. (GB)Number of authors 12 Source Title Acta Crystallographica Section D-Structural Biology. - : Oxford Blackwell - ISSN 2059-7983
Roč. 72, č. 2 (2016), s. 254-265Number of pages 12 s. Language eng - English Country GB - United Kingdom Keywords cellulose degradation ; N-glycan ; biofuels ; glycoblocks Subject RIV EB - Genetics ; Molecular Biology R&D Projects ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Institutional support BTO-N - RVO:86652036 UT WOS 000373718400008 DOI 10.1107/S2059798315024237 Annotation The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. These technologies harness diverse plant degrading enzymes, classical exo- and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the beta-D-linked polysaccharide. A major drawback with this process is that the exo-acting cellobiohydrolases suffer from severe inhibition from their cellobiose product. beta-D-Glucosidases are therefore important for liberating glucose from cellobiose and thereby relieving limiting product inhibition. Here, the three-dimensional structures of two industrially important family GH3 beta-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 angstrom resolution, are reported. Both enzymes display a three-domain structure with the catalytic domain at the interface, as originally shown for barley beta-D-glucan exohydrolase. Both enzymes show extensive N-glycosylation, with only a few external sites being truncated to a single GlcNAc molecule. Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. The extensive glycans pose special problems for crystallographic refinement, and new techniques and protocols were developed especially for this work. These protocols ensured that all of the d-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy C-4(1) chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal beta-D-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application. Workplace Institute of Biotechnology Contact Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700 Year of Publishing 2017
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