Computational design of the temperature optimum of an enzyme reaction
Cold-adapted enzymes are characterized both by a higher catalytic activity at low temperatures and by having their temperature optimum down-shifted, compared to mesophilic orthologs. In several cases, the optimum does not coincide with the onset of protein melting but reflects some other type of ina...
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American Association for the Advancement of Science (AAAS)
2023
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Online Access: | http://dx.doi.org/10.1126/sciadv.adi0963 https://www.science.org/doi/pdf/10.1126/sciadv.adi0963 |
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craaas:10.1126/sciadv.adi0963 2024-09-15T17:40:10+00:00 Computational design of the temperature optimum of an enzyme reaction van der Ent, Florian Skagseth, Susann Lund, Bjarte A. Sočan, Jaka Griese, Julia J. Brandsdal, Bjørn O. Åqvist, Johan 2023 http://dx.doi.org/10.1126/sciadv.adi0963 https://www.science.org/doi/pdf/10.1126/sciadv.adi0963 en eng American Association for the Advancement of Science (AAAS) Science Advances volume 9, issue 26 ISSN 2375-2548 journal-article 2023 craaas https://doi.org/10.1126/sciadv.adi0963 2024-08-01T04:01:11Z Cold-adapted enzymes are characterized both by a higher catalytic activity at low temperatures and by having their temperature optimum down-shifted, compared to mesophilic orthologs. In several cases, the optimum does not coincide with the onset of protein melting but reflects some other type of inactivation. In the psychrophilic α-amylase from an Antarctic bacterium, the inactivation is thought to originate from a specific enzyme-substrate interaction that breaks around room temperature. Here, we report a computational redesign of this enzyme aimed at shifting its temperature optimum upward. A set of mutations designed to stabilize the enzyme-substrate interaction were predicted by computer simulations of the catalytic reaction at different temperatures. The predictions were verified by kinetic experiments and crystal structures of the redesigned α-amylase, showing that the temperature optimum is indeed markedly shifted upward and that the critical surface loop controlling the temperature dependence approaches the target conformation observed in a mesophilic ortholog. Article in Journal/Newspaper Antarc* Antarctic AAAS Resource Center (American Association for the Advancement of Science) Science Advances 9 26 |
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Open Polar |
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AAAS Resource Center (American Association for the Advancement of Science) |
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language |
English |
description |
Cold-adapted enzymes are characterized both by a higher catalytic activity at low temperatures and by having their temperature optimum down-shifted, compared to mesophilic orthologs. In several cases, the optimum does not coincide with the onset of protein melting but reflects some other type of inactivation. In the psychrophilic α-amylase from an Antarctic bacterium, the inactivation is thought to originate from a specific enzyme-substrate interaction that breaks around room temperature. Here, we report a computational redesign of this enzyme aimed at shifting its temperature optimum upward. A set of mutations designed to stabilize the enzyme-substrate interaction were predicted by computer simulations of the catalytic reaction at different temperatures. The predictions were verified by kinetic experiments and crystal structures of the redesigned α-amylase, showing that the temperature optimum is indeed markedly shifted upward and that the critical surface loop controlling the temperature dependence approaches the target conformation observed in a mesophilic ortholog. |
format |
Article in Journal/Newspaper |
author |
van der Ent, Florian Skagseth, Susann Lund, Bjarte A. Sočan, Jaka Griese, Julia J. Brandsdal, Bjørn O. Åqvist, Johan |
spellingShingle |
van der Ent, Florian Skagseth, Susann Lund, Bjarte A. Sočan, Jaka Griese, Julia J. Brandsdal, Bjørn O. Åqvist, Johan Computational design of the temperature optimum of an enzyme reaction |
author_facet |
van der Ent, Florian Skagseth, Susann Lund, Bjarte A. Sočan, Jaka Griese, Julia J. Brandsdal, Bjørn O. Åqvist, Johan |
author_sort |
van der Ent, Florian |
title |
Computational design of the temperature optimum of an enzyme reaction |
title_short |
Computational design of the temperature optimum of an enzyme reaction |
title_full |
Computational design of the temperature optimum of an enzyme reaction |
title_fullStr |
Computational design of the temperature optimum of an enzyme reaction |
title_full_unstemmed |
Computational design of the temperature optimum of an enzyme reaction |
title_sort |
computational design of the temperature optimum of an enzyme reaction |
publisher |
American Association for the Advancement of Science (AAAS) |
publishDate |
2023 |
url |
http://dx.doi.org/10.1126/sciadv.adi0963 https://www.science.org/doi/pdf/10.1126/sciadv.adi0963 |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
Science Advances volume 9, issue 26 ISSN 2375-2548 |
op_doi |
https://doi.org/10.1126/sciadv.adi0963 |
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Science Advances |
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9 |
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26 |
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1810485235602685952 |