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Kuhaudomlarp, Sakonwan, Stevenson, Clare E.M., Lawson, David M. and Field, Robert A. 
ORCID: https://orcid.org/0000-0001-8574-0275
(2019)
The structure of a GH149 β-(1 → 3) glucan phosphorylase reveals a new surface oligosaccharide binding site and additional domains that are absent in the disaccharide-specific GH94 glucose-β-(1 → 3)-glucose (laminaribiose) phosphorylase.
Proteins: Structure, Function and Bioinformatics, 87 (10).
pp. 885-892.
ISSN 0887-3585
Abstract
Glycoside phosphorylases (GPs) with specificity for β-(1 → 3)-gluco-oligosaccharides are potential candidate biocatalysts for oligosaccharide synthesis. GPs with this linkage specificity are found in two families thus far—glycoside hydrolase family 94 (GH94) and the recently discovered glycoside hydrolase family 149 (GH149). Previously, we reported a crystallographic study of a GH94 laminaribiose phosphorylase with specificity for disaccharides, providing insight into the enzyme's ability to recognize its' sugar substrate/product. In contrast to GH94, characterized GH149 enzymes were shown to have more flexible chain length specificity, with preference for substrate/product with higher degree of polymerization. In order to advance understanding of the specificity of GH149 enzymes, we herein solved X-ray crystallographic structures of GH149 enzyme Pro_7066 in the absence of substrate and in complex with laminarihexaose (G6). The overall domain organization of Pro_7066 is very similar to that of GH94 family enzymes. However, two additional domains flanking its catalytic domain were found only in the GH149 enzyme. Unexpectedly, the G6 complex structure revealed an oligosaccharide surface binding site remote from the catalytic site, which, we suggest, may be associated with substrate targeting. As such, this study reports the first structure of a GH149 phosphorylase enzyme acting on β-(1 → 3)-gluco-oligosaccharides and identifies structural elements that may be involved in defining the specificity of the GH149 enzymes.
| Item Type: | Article |
|---|---|
| Additional Information: | Funding Information: Biotechnology and Biological Sciences Research Council, Grant/Award Numbers: BB/LO1413/1, BB/PO12523/1; FP7 Ideas: European Research Council, Grant/Award Number: 283570; Royal Thai government scholarship; European Community's Seventh Framework Program, Grant/Award Number: 283570; Royal Thai Government Scholarship program; John Innes Foundation; Open Plant Synthetic Biology Centre, Grant/Award Number: BB/LO1413/1; UK BBSRC Institute Strategic Program Molecules from Nature (MfN), Grant/Award Number: BB/PO12523/1 Funding Information: This work was supported by the UK BBSRC Institute Strategic Program Molecules from Nature (MfN) (BB/PO12523/1); the Open Plant Synthetic Biology Centre (BB/LO1413/1); the John Innes Foundation; and the Royal Thai Government Scholarship program. We acknowledge the Diamond Light Source for access to beamlines I03 and I04 under proposal MX13467 with support from the European Community's Seventh Framework Program (FP7/2007-2013) under Grant Agreement 283570 (BioStruct-X). Publisher Copyright: © 2019 The Authors. Proteins: Structure, Function, and Bioinformatics published by Wiley Periodicals, Inc. |
| Uncontrolled Keywords: | carbohydrate-active enzyme,glycoside phosphorylase,oligosaccharide,surface binding site,structural biology,biochemistry,molecular biology ,/dk/atira/pure/subjectarea/asjc/1300/1315 |
| Faculty \ School: | Faculty of Science > School of Biological Sciences Faculty of Science > School of Chemistry, Pharmacy and Pharmacology |
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| Depositing User: | LivePure Connector |
| Date Deposited: | 02 Sep 2024 15:30 |
| Last Modified: | 25 Sep 2024 18:05 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/96445 |
| DOI: | 10.1002/prot.25745 |
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