Rational Enzyme Design without Structural Knowledge : A Sequence-Based Approach for Efficient Generation of Transglycosylases
(2021) In Chemistry: A European Journal 27(40). p.10323-10334- Abstract
Glycobiology is dogged by the relative scarcity of synthetic, defined oligosaccharides. Enzyme-catalysed glycosylation using glycoside hydrolases is feasible but is hampered by the innate hydrolytic activity of these enzymes. Protein engineering is useful to remedy this, but it usually requires prior structural knowledge of the target enzyme, and/or relies on extensive, time-consuming screening and analysis. Here we describe a straightforward strategy that involves rational rapid in silico analysis of protein sequences. The method pinpoints 6-12 single mutant candidates to improve transglycosylation yields. Requiring very little prior knowledge of the target enzyme other than its sequence, the method is generic and procures catalysts... (More)
Glycobiology is dogged by the relative scarcity of synthetic, defined oligosaccharides. Enzyme-catalysed glycosylation using glycoside hydrolases is feasible but is hampered by the innate hydrolytic activity of these enzymes. Protein engineering is useful to remedy this, but it usually requires prior structural knowledge of the target enzyme, and/or relies on extensive, time-consuming screening and analysis. Here we describe a straightforward strategy that involves rational rapid in silico analysis of protein sequences. The method pinpoints 6-12 single mutant candidates to improve transglycosylation yields. Requiring very little prior knowledge of the target enzyme other than its sequence, the method is generic and procures catalysts for the formation of glycosidic bonds involving various d / l -, α/β-pyranosides or furanosides, and exo - and endo -action. Moreover, mutations validated in one enzyme can be transposed to others, even distantly related enzymes.
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- author
- organization
- publishing date
- 2021-04-29
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Chemistry: A European Journal
- volume
- 27
- issue
- 40
- pages
- 10323 - 10334
- publisher
- Wiley-Blackwell
- external identifiers
-
- scopus:85107290185
- pmid:33914359
- ISSN
- 1521-3765
- DOI
- 10.1002/chem.202100110
- language
- English
- LU publication?
- yes
- id
- b604f553-a061-4dff-a8b2-636f3cd66acd
- date added to LUP
- 2021-05-26 22:12:24
- date last changed
- 2024-11-18 03:58:53
@article{b604f553-a061-4dff-a8b2-636f3cd66acd, abstract = {{<p>Glycobiology is dogged by the relative scarcity of synthetic, defined oligosaccharides. Enzyme-catalysed glycosylation using glycoside hydrolases is feasible but is hampered by the innate hydrolytic activity of these enzymes. Protein engineering is useful to remedy this, but it usually requires prior structural knowledge of the target enzyme, and/or relies on extensive, time-consuming screening and analysis. Here we describe a straightforward strategy that involves rational rapid in silico analysis of protein sequences. The method pinpoints 6-12 single mutant candidates to improve transglycosylation yields. Requiring very little prior knowledge of the target enzyme other than its sequence, the method is generic and procures catalysts for the formation of glycosidic bonds involving various d / l -, α/β-pyranosides or furanosides, and <i>exo </i>- and <i>endo </i>-action. Moreover, mutations validated in one enzyme can be transposed to others, even distantly related enzymes.</p>}}, author = {{Teze, David and Zhao, Jiao and Wiemann, Mathias and Kazi, Zubaida G. A. and Lupo, Rossana and Zeuner, Birgitte and Vuillemin, Marlène and Rønne, Mette E. and Carlström, Göran and Duus, Jens Ø. and Sanejouand, Yves-Henri and O'Donohue, Michael J. and Karlsson, Eva Nordberg and Fauré, Régis and Stålbrand, Henrik and Svensson, Birte}}, issn = {{1521-3765}}, language = {{eng}}, month = {{04}}, number = {{40}}, pages = {{10323--10334}}, publisher = {{Wiley-Blackwell}}, series = {{Chemistry: A European Journal}}, title = {{Rational Enzyme Design without Structural Knowledge : A Sequence-Based Approach for Efficient Generation of Transglycosylases}}, url = {{http://dx.doi.org/10.1002/chem.202100110}}, doi = {{10.1002/chem.202100110}}, volume = {{27}}, year = {{2021}}, }