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Open AccessArticle

Structural Comparison of a Promiscuous and a Highly Specific Sucrose 6F-Phosphate Phosphorylase

1
Centre for Synthetic Biology (CSB), Department of Biotechnology, Ghent University, Coupure Links 653, 9000 Ghent, Belgium
2
Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2019, 20(16), 3906; https://doi.org/10.3390/ijms20163906
Received: 17 July 2019 / Revised: 6 August 2019 / Accepted: 8 August 2019 / Published: 11 August 2019
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Abstract

In family GH13 of the carbohydrate-active enzyme database, subfamily 18 contains glycoside phosphorylases that act on α-sugars and glucosides. Because their phosphorolysis reactions are effectively reversible, these enzymes are of interest for the biocatalytic synthesis of various glycosidic compounds. Sucrose 6F-phosphate phosphorylases (SPPs) constitute one of the known substrate specificities. Here, we report the characterization of an SPP from Ilumatobacter coccineus with a far stricter specificity than the previously described promiscuous SPP from Thermoanaerobacterium thermosaccharolyticum. Crystal structures of both SPPs were determined to provide insight into their similarities and differences. The residues responsible for binding the fructose 6-phosphate group in subsite +1 were found to differ considerably between the two enzymes. Furthermore, several variants that introduce a higher degree of substrate promiscuity in the strict SPP from I. coccineus were designed. These results contribute to an expanded structural knowledge of enzymes in subfamily GH13_18 and facilitate their rational engineering. View Full-Text
Keywords: GH13_18; sucrose phosphorylase; glycoside phosphorylase; Ilumatobacter coccineus; Thermoanaerobacterium thermosaccharolyticum; crystallography GH13_18; sucrose phosphorylase; glycoside phosphorylase; Ilumatobacter coccineus; Thermoanaerobacterium thermosaccharolyticum; crystallography
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Franceus, J.; Capra, N.; Desmet, T.; Thunnissen, A.-M.W. Structural Comparison of a Promiscuous and a Highly Specific Sucrose 6F-Phosphate Phosphorylase. Int. J. Mol. Sci. 2019, 20, 3906.

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