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Development and application of a highly α2,6-selective pseudosialidase

preprint
submitted on 23.08.2017 and posted on 31.08.2017 by Peter Both, Michel Riese, Christopher J. Gray, Kun Huang, Edward G. Pallister, Iaroslav Kosov, Louis P. Conway, Josef Voglmeir, Sabine L. Flitsch

In this manuscript we address an important gap in our current carbohydrate active enzyme toolbox, by developing a highly a2,6-selective (over a2,3-selective) de facto sialidase that is necessary both for glycan analysis and glycoconjugate remodeling. Both glycosidic linkages are commonly found in animal biology and each has been shown to have distinct biological function.

Our approach is novel in that it harnesses the high selectivity of known glycosyltransferases ‘in reverse’ for effective hydrolysis, converting transferases to hydrolases by reaction engineering.

More specifically, we demonstrate that the a2,6-specific pseudosialidase activity of Photobacterium sp. JT-ISH-224 a2,6-sialyltransferase can be used effectively for highly a2,6 selective hydrolysis on a broad range of analytes: small synthetic probes, isolated complex glycans and complex mixtures of glycoproteins.

Funding

This study was supported by the BBSRC, EPSRC, InnovateUK: IBCatalyst [BB/M02903411] and by the European Union’s Sev-enth Framework Programme [FP7/2007–2013] under grant agreement no 266025.

History

Topic

  • Chemical Biology

Email Address of Submitting Author

peter.both@manchester.ac.uk

Email Address(es) for Other Author(s)

Michel RIESE (michel.riese@postgrad.manchester.ac.uk) Christopher J. GRAY (christopher.gray@manchester.ac.uk) Kun HUANG (kun.huang-3@manchester.ac.uk) Edward G. Pallister (edward.pallister@postgrad.manchester.ac.uk) Iaroslav KOSOV (iaroslav.kosov@student.manchester.ac.uk) Louis P. CONWAY (louis.conway@njau.edu.cn) Josef VOGLMEIR (josef.voglmeir@njau.edu.cn) Sabine L. FLITSCH (sabine.flitsch@manchester.ac.uk)

Institution

Manchester Institute of Biotechnology, The University of Manchester

Country

United Kingdom

ORCID For Submitting Author

0000-0002-4969-9014

Declaration of Conflict of Interest

no conflict of interest

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in Glycobiology

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