Toward the design of efficient transglycosidases: the case of the GH1 of Thermus thermophilus
Autor: | Philippe Arnaud, Yves-Henri Sanejouand, Benoit David, Charles Tellier |
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Přispěvatelé: | Unité de fonctionnalité et ingénierie de protéines (UFIP), Université de Nantes - UFR des Sciences et des Techniques (UN UFR ST), Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS) |
Jazyk: | angličtina |
Rok vydání: | 2019 |
Předmět: |
Glycosylation
Glycoside Hydrolases Stereochemistry Protein Conformation Mutant Bioengineering Molecular Dynamics Simulation 010402 general chemistry Protein Engineering 01 natural sciences Biochemistry Substrate Specificity Turn (biochemistry) 03 medical and health sciences Glycoside hydrolase [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Molecular Biology ComputingMilieux_MISCELLANEOUS 030304 developmental biology 0303 health sciences biology Chemistry Thermus thermophilus Wild type [CHIM.CATA]Chemical Sciences/Catalysis Hydrogen-Ion Concentration biology.organism_classification 0104 chemical sciences Kinetics Yield (chemistry) Mutation Biocatalysis Biotechnology |
Zdroj: | Protein Engineering, Design and Selection Protein Engineering, Design and Selection, Oxford University Press (OUP), 2019, 32 (7), pp.309-316. ⟨10.1093/protein/gzz032⟩ |
ISSN: | 1741-0126 1741-0134 |
Popis: | Using the information available in the sequences of well-characterized transglycosidases found in plants, mutations were introduced in the glycoside hydrolase of the bacterium Thermus thermophilus, with the aim of turning it into an efficient transglycosidase. All mutants happen to have fair catalytic efficiencies, being at worst 25 times less efficient than the wild type. Noteworthy, W120F, one of our high transglycosylation yield (≈ 50%) mutants, is only two times less efficient than the wild type. Interestingly, while in the wild type the sidechain of the acid–base is only found able to sample a pair of equivalent conformations during 0.5-μs-long molecular dynamics simulations, its flexibility is much higher in the case of the high transglycosylation yield mutants. Our results thus suggest that engineering the flexibility of the acid–base of a retaining glycoside hydrolase could be a general way to turn it into an efficient transglycosidase. |
Databáze: | OpenAIRE |
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