A single point mutation alters the transglycosylation/hydrolysis partition, significantly enhancing the synthetic capability of an endo-glycoceramidase
| Autor: | Julien Durand, Michael J. O’Donohue, Antoni Planas, Laurie Watterlot, Sophie Bozonnet, Xevi Biarnés, Cyrielle Bonzom, Vinciane Borsenberger, Régis Fauré |
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| Přispěvatelé: | Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Laboratory of Biochemistry, Institut Químic de Sarrià, Universitat Ramon Llull [Barcelona] (URL), ERA-NET BIOSUR (0315928A ERA-IB10.039), Region Midi-Pyrenees grant DESR-Recherche (14052246), MINECO of Spain (BIO2013-49022-C2-1-R), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA) |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification Ketone Stereochemistry [SDV]Life Sciences [q-bio] Mutant alkyl cellobiosides General Chemistry Acceptor Catalysis Transition state 03 medical and health sciences Hydrolysis chemoenzymatic glycosynthesis 030104 developmental biology chemistry Glycoside hydrolase glycoside hydrolase Alkyl mutagenesis transglycosylation molecular interactions |
| Zdroj: | ACS Catalysis ACS Catalysis, 2016, 6 (12), pp.8264-8275. ⟨10.1021/acscatal.6b02159⟩ ACS Catalysis, American Chemical Society, 2016, 6 (12), pp.8264-8275. ⟨10.1021/acscatal.6b02159⟩ |
| ISSN: | 2155-5435 |
| Popis: | 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 beta-cellobioside, have provided appropriate conditions for the efficient synthesis of alkyl beta-cellobioside derivatives. The mutant D311Y was characterized by a lowered KM value for the hydrolysis of 2-chloro-4-nitrophenyl beta-cellobioside and increased transglycosylation when using aliphatic 1,3-diols or alcohols bearing a delta-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. |
| Databáze: | OpenAIRE |
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