An Epoxide Intermediate in Glycosidase Catalysis

Lukasz F Sobala; Gaetano Speciale; Sha Zhu; Lluis Raich; Natalia Sannikova; Andrew J Thompson; Zalihe Hakki; Dan Lu; Saeideh Shamsi Kazem Abadi; Andrew R Lewis; Victor Rojas-Cervellera; Ganeko Bernardo-Seisdedos; Yongmin Zhang; Oscar Millet; Jesus Jimenez-Barbero; Andrew J Bennet; Matthieu Sollogoub; Carme Rovira; Gideon J Davies; Spencer J Williams

Journal article

An Epoxide Intermediate in Glycosidase Catalysis

Lukasz F Sobala, Gaetano Speciale, Sha Zhu, Lluis Raich, Natalia Sannikova, Andrew J Thompson, Zalihe Hakki, Dan Lu, Saeideh Shamsi Kazem Abadi, Andrew R Lewis, Victor Rojas-Cervellera, Ganeko Bernardo-Seisdedos, Yongmin Zhang, Oscar Millet, Jesus Jimenez-Barbero, Andrew J Bennet, Matthieu Sollogoub, Carme Rovira, Gideon J Davies, Spencer J Williams

ACS CENTRAL SCIENCE | AMER CHEMICAL SOC | Published : 2020

DOI: 10.1021/acscentsci.0c00111

Abstract

Retaining glycoside hydrolases cleave their substrates through stereochemical retention at the anomeric position. Typically, this involves two-step mechanisms using either an enzymatic nucleophile via a covalent glycosyl enzyme intermediate or neighboring-group participation by a substrate-borne 2-acetamido neighboring group via an oxazoline intermediate; no enzymatic mechanism with participation of the sugar 2-hydroxyl has been reported. Here, we detail structural, computational, and kinetic evidence for neighboring-group participation by a mannose 2-hydroxyl in glycoside hydrolase family 99 endo-α-1,2-mannanases. We present a series of crystallographic snapshots of key species along the re..

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University of Melbourne Researchers

Spencer Williams Author Chemistry

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Grants

Awarded by Australian Research Council

Awarded by European Research Council

Awarded by Natural Sciences and Engineering Research Council of Canada

Awarded by Spanish Ministry of Science, Innovation and Universities (MICINN/AEI/FEDER)

Awarded by Spanish Structures of Excellence Maria de Maeztu

Awarded by Agency for Management of University and Research Grants of Generalitat de Catalunya (AGAUR)

Awarded by Agencia Estatal de Investigacion of Spain

Awarded by European Research Council (RECGLYCANMR, ERC AdG)

Awarded by Severo Ochoa Excellence accreditation

Funding Acknowledgements

This work was supported by the Australian Research Council (FT130100103, DP120101396, DP180101957), the BBSRC (A.J.T.), European Research Council (ERC-2012-AdG-322942 "Glycopoise"; L. F.S.), Natural Sciences and Engineering Research Council of Canada (A.J.B. Discovery Grant: 201704910), the Spanish Ministry of Science, Innovation and Universities (MICINN/AEI/FEDER) (CTQ2017-85496-P to C.R.), the Spanish Structures of Excellence Maria de Maeztu (MDM-2017-0767 to C.R.), and the Agency for Management of University and Research Grants of Generalitat de Catalunya (AGAUR) (2017SGR-1189 to C.R.). G.J.D. is funded by the Royal Society "Ken Murray" Professorship. We are grateful to Sivanandam Veeramuthu for help with NMR. We thank Diamond Light Source for access to beamlines I02, I04, and I04-1 (proposals mx-9948 and mx-13587). In-house crystal screening was performed on X-ray equipment provided, in part, by the Wellcome Trust. The authors gratefully acknowledge the computer resources and technical support provided by the Barcelona Supercomputing Center (BSC-CNS, Barcelona, Spain). The group at Bilbao thanks the Agencia Estatal de Investigacion of Spain (Grants RTI2018-101269-BI00 and RTI2018-094751-B-C21), the European Research Council (RECGLYCANMR, ERC AdG 788143), and the Severo Ochoa Excellence accreditation (SEV2016-0644).