Rational Enzyme Design Without Structural Knowledge: A Sequence-Based Approach for Efficient Generation of Glycosylation Catalysts
| Autor: | Birte Svensson, Henrik Stålbrand, Eva Karlsson, Rossana Lupo, Jens Ø. Duus, David Teze, Régis Fauré, Yves-Henri Sanejouand, Zubaida Gulshan Kazi, Michael J. O’Donohue, Mathias Wiemann, Göran Carlström, Mette Errebo Rønne, Jiao Zhao |
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| Přispěvatelé: | Technical University of Denmark [Lyngby] (DTU), Toulouse Biotechnology Institute (TBI), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Lund University [Lund], Centre for Analysis and Synthesis, Department of Chemistry, Lund University, Université de Nantes (UN), Department of Biotechnology and Biomedicine, Department of biochemistry and structural biology, Biotechnology, Department of Chemistry, Department of Chemistry, Technical University of Denmark, Lyngby, Unité de fonctionnalité et ingénierie de protéines (UFIP), Université de Nantes - Faculté des Sciences et des Techniques, Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2020 |
| Předmět: |
chemistry.chemical_classification
2. Zero hunger [SDV.BIO]Life Sciences [q-bio]/Biotechnology Glycosylation [CHIM.ORGA]Chemical Sciences/Organic chemistry 010405 organic chemistry Chemistry Rational design [CHIM.CATA]Chemical Sciences/Catalysis Protein engineering 010402 general chemistry 01 natural sciences [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biomolecules [q-bio.BM] 0104 chemical sciences Catalysis chemistry.chemical_compound Enzyme Biochemistry Biocatalysis [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Glycoside hydrolase Sequence (medicine) |
| Popis: | We present an enzyme engineering approach based solely on amino-acids sequence to convert glycoside hydrolases into transglycosylases. We demonstrate its effectiveness on enzymes form five different glycoside hydrolase families, synthesizing various oligosaccharides containing different α-/β-pyranosides or furanosides in one-step with high yields.; Glycobiology is dogged by the relative scarcity of synthetic, defined oligosaccha-9 rides. Enzyme-catalysed glycosylation using glycoside hydrolases is feasible, but is 10 hampered by the innate hydrolytic activity of these enzymes. Protein engineering 11 methods are applicable, though usually require prior structural knowledge of the 12 target enzyme and the use of powerful computing methods, and/or relies on extensive 13 screening methodologies. Here we describe a straightforward strategy that involves 14 rapid in silico analysis of protein sequences. The method pinpoints a small number 15 ( |
| Databáze: | OpenAIRE |
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