@article{8b606d970cfe4732a2c5def9653799fe,
title = "A single point mutation alters the transglycosylation/Hydrolysis partition, significantly enhancing the synthetic capability of an endo-Glycoceramidase",
abstract = "The mutation of D311 to tyrosine in endo-glycoceramidase II from Rhodococcus sp. and the use of a poorly recognized substrate, 2-chloro-4-nitrophenyl β-cellobioside, have provided appropriate conditions for the efficient synthesis of alkyl β-cellobioside derivatives. The mutant D311Y was characterized by a lowered KM value for the hydrolysis of 2-chloro-4-nitrophenyl β-cellobioside and increased transglycosylation when using aliphatic 1,3-diols or alcohols bearing a δ-hydroxy ketone function as acceptors. Closer analysis revealed that the transglycosylation/hydrolysis ratio in reactions catalyzed by the mutant was completely inversed and weak secondary hydrolysis was postponed, thus providing the basis for high transglycosylation yields (between 68 and 93%). Overall, results confirm that the enhancement of transglycosylation in glycoside hydrolases can be achieved by a combination of destabilized transition states and increased recognition for acceptor molecules.",
keywords = "Alkyl cellobiosides, Chemoenzymatic glycosynthesis, Glycoside hydrolase, Molecular interactions, Mutagenesis, Transglycosylation",
author = "Julien Durand and Xevi Biarnes and Laurie Watterlot and Cyrielle Bonzom and Vinciane Borsenberger and Antoni Planas and Sophie Bozonnet and O{\textquoteright}Donohue, {Michael J.} and Regis Faure",
note = "Funding Information: This work was performed in the framework of ERA-NET BIOSURF (grant no. 0315928A, ERA-IB10.039). NMR analyses were performed using facilities at MetaToul (Metabolomics & Fluxomics Facitilies, Toulouse, France, www.metatoul.fr), which is part of the national infrastructure MetaboHUB (The French National infrastructure for metab-olomics and fluxomics, www.metabohub.fr) and is supported by grants from the Reǵ ion Midi-Pyre{\'n}e{\'e}s, the European Regional Development Fund, SICOVAL, IBiSa-France, CNRS, and INRA. Work requiring automated liquid handling was performed using the ICEO facility that is part of the PICT infrastructure and is financially supported by IBiSa-France, CNRS, and INRA. The contribution from R.F. was partially supported by the Reǵ ion Midi-Pyre{\'n}e{\'e}s grant DESR-Recherche 14052246 (CTP-B) and the research mobility grants from INSA Toulouse (2015). A.P. acknowledges grant BIO2013-49022-C2-1-R from the MINECO of Spain. The authors thank Prof. S. Cottaz and Dr. S. Fort (CERMAV-CNRS) for the gift of β-cellobiosyl fluoride heptaacetate and Prof. S. G. Withers (Univ. of British Columbia) for insightful discussions. Funding Information: This work was performed in the framework of ERA-NET BIOSURF (grant no. 0315928A, ERA-IB10.039). NMR analyses were performed using facilities at MetaToul (Metabolomics & Fluxomics Facitilies, Toulouse, France, www.metatoul.fr), which is part of the national infrastructure MetaboHUB (The French National infrastructure for metab-olomics and fluxomics, www.metabohub.fr) and is supported by grants from the Region Midi-Pyrene {\'e}s, the European Regional Development Fund, SICOVAL, IBiSa-France, CNRS, and INRA. Work requiring automated liquid handling was performed using the ICEO facility that is part of the PICT infrastructure and is financially supported by IBiSa-France, CNRS, and INRA. The contribution from R.F. was partially supported by the Region Midi-Pyrene {\'e}s grant DESR-Recherche 14052246 (CTP-B) and the research mobility grants from INSA Toulouse (2015). A.P. acknowledges grant BIO2013-49022-C2-1-R from the MINECO of Spain. The authors thank Prof. S. Cottaz and Dr. S. Fort (CERMAV-CNRS) for the gift of β-cellobiosyl fluoride heptaacetate and Prof. S. G. Withers (Univ. of British Columbia) for insightful discussions. Publisher Copyright: {\textcopyright} 2016 American Chemical Society.",
year = "2016",
month = dec,
day = "2",
doi = "10.1021/acscatal.6b02159",
language = "English",
volume = "6",
pages = "8264--8275",
journal = "ACS Catalysis",
issn = "2155-5435",
publisher = "American Chemical Society",
number = "12",
}