Molecular recognition of natural and non‐natural substrates by cellodextrin phosphorylase from Ruminiclostridium thermocellum investigated by NMR spectroscopy

Gabrielli, Valeria, Muñoz-García, Juan Carlos ORCID: https://orcid.org/0000-0003-2246-3236, Pergolizzi, Giulia, De Andrade, Peterson, Khimyak, Yaroslav ORCID: https://orcid.org/0000-0003-0424-4128, Field, Robert A and Angulo, Jesus ORCID: https://orcid.org/0000-0001-7250-5639 (2021) Molecular recognition of natural and non‐natural substrates by cellodextrin phosphorylase from Ruminiclostridium thermocellum investigated by NMR spectroscopy. Chemistry – A European Journal, 27 (63). pp. 15688-15698. ISSN 0947-6539

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Abstract

β-1→4-Glucan polysaccharides like cellulose, derivatives and analogues, are attracting attention due to their unique physicochemical properties, as ideal candidates for many different applications in biotechnology. Access to these polysaccharides with a high level of purity at scale is still challenging, and eco-friendly alternatives by using enzymes in vitro are highly desirable. One prominent candidate enzyme is cellodextrin phosphorylase (CDP) from Ruminiclostridium thermocellum, which is able to yield cellulose oligomers from short cellodextrins and α-d-glucose 1-phosphate (Glc-1-P) as substrates. Remarkably, its broad specificity towards donors and acceptors allows the generation of highly diverse cellulose-based structures to produce novel materials. However, to fully exploit this CDP broad specificity, a detailed understanding of the molecular recognition of substrates by this enzyme in solution is needed. Herein, we provide a detailed investigation of the molecular recognition of ligands by CDP in solution by saturation transfer difference (STD) NMR spectroscopy, tr-NOESY and protein-ligand docking. Our results, discussed in the context of previous reaction kinetics data in the literature, allow a better understanding of the structural basis of the broad binding specificity of this biotechnologically relevant enzyme.

Item Type: Article
Additional Information: Funding Information: We thank the BBSRC for provision of Norwich Research Park Bioscience Doctoral Training Grant (BB/M011216/1) to V.G. J.A. acknowledges funding support from the Spanish Ministry of Science and Innovation through the grant PID2019‐109395GB‐I00, and the Biotechnology and Biological Sciences Research Council (BBSRC; BB/P010660/1). The Engineering and Physical Sciences Research Council (EPSRC) is acknowledged for provision of financial support (EP/N033337/1) for J.C.M.G., J.A. and Y.Z.K. We are also grateful for UEA Faculty of Science NMR facility. Work at the John Innes Centre was supported by the UK BBSRC Institute Strategic Program on Molecules from Nature ‐ Products and Pathways [BBS/E/J/000PR9790] and the John Innes Foundation, the BBSRC, EPSRC, and InnovateUK: IBCatalyst (GrantBB/M02903411). We would also like to acknowledge the support of the GelEnz consortium, which is funded by EPSRC (Grant Research Number: IUK 4159000 442149). Additional research data supporting this publication are available as electronic supplementary files at the following link: XXX. Publisher Copyright: © 2021 The Authors. Chemistry - A European Journal published by Wiley-VCH GmbH
Uncontrolled Keywords: cellodextrin phosphorylase,ligand-based nmr spectroscopy,molecular docking,protein-ligand interactions,catalysis,organic chemistry ,/dk/atira/pure/subjectarea/asjc/1500/1503
Faculty \ School: Faculty of Science > School of Pharmacy
Faculty of Science > School of Biological Sciences
Faculty of Science > School of Chemistry
Related URLs:
Depositing User: LivePure Connector
Date Deposited: 03 Sep 2021 00:06
Last Modified: 22 Oct 2022 13:30
URI: https://ueaeprints.uea.ac.uk/id/eprint/81281
DOI: 10.1002/chem.202102039

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