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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Uppsala universitet, Beräkningsbiologi och bioinformatik
2023
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ftuppsalauniv:oai:DiVA.org:uu-508544 2023-12-24T10:11:19+01:00 Computational design of the temperature optimum of an enzyme reaction van der Ent, Florian Skagseth, Susann Lund, Bjarte A. Socan, Jaka Griese, Julia J. Brandsdal, Bjorn O. Åqvist, Johan 2023 application/pdf http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-508544 https://doi.org/10.1126/sciadv.adi0963 eng eng Uppsala universitet, Beräkningsbiologi och bioinformatik Uppsala universitet, Institutionen för cell- och molekylärbiologi Uppsala universitet, Strukturbiologi Univ Tromso, Arctic Univ Norway, Hylleraas Ctr Quantum Mol Sci, Dept Chem, N-9037 Tromso, Norway. Natl Inst Chem, SI-1001 Ljubljana, Slovenia. American Association for the Advancement of Science (AAAS) Science Advances, 2023, 9:26, orcid:0000-0002-0933-4547 orcid:0000-0001-7757-7172 orcid:0000-0003-3686-3062 orcid:0000-0003-2091-0610 http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-508544 doi:10.1126/sciadv.adi0963 PMID 37379391 ISI:001029314100001 info:eu-repo/semantics/openAccess Biochemistry and Molecular Biology Biokemi och molekylärbiologi Article in journal info:eu-repo/semantics/article text 2023 ftuppsalauniv https://doi.org/10.1126/sciadv.adi0963 2023-11-29T23:31:59Z 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 a-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 a-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 Uppsala University: Publications (DiVA) Antarctic Science Advances 9 26 |
institution |
Open Polar |
collection |
Uppsala University: Publications (DiVA) |
op_collection_id |
ftuppsalauniv |
language |
English |
topic |
Biochemistry and Molecular Biology Biokemi och molekylärbiologi |
spellingShingle |
Biochemistry and Molecular Biology Biokemi och molekylärbiologi van der Ent, Florian Skagseth, Susann Lund, Bjarte A. Socan, Jaka Griese, Julia J. Brandsdal, Bjorn O. Åqvist, Johan Computational design of the temperature optimum of an enzyme reaction |
topic_facet |
Biochemistry and Molecular Biology Biokemi och molekylärbiologi |
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 a-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 a-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. Socan, Jaka Griese, Julia J. Brandsdal, Bjorn O. Åqvist, Johan |
author_facet |
van der Ent, Florian Skagseth, Susann Lund, Bjarte A. Socan, Jaka Griese, Julia J. Brandsdal, Bjorn 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 |
Uppsala universitet, Beräkningsbiologi och bioinformatik |
publishDate |
2023 |
url |
http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-508544 https://doi.org/10.1126/sciadv.adi0963 |
geographic |
Antarctic |
geographic_facet |
Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_relation |
Science Advances, 2023, 9:26, orcid:0000-0002-0933-4547 orcid:0000-0001-7757-7172 orcid:0000-0003-3686-3062 orcid:0000-0003-2091-0610 http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-508544 doi:10.1126/sciadv.adi0963 PMID 37379391 ISI:001029314100001 |
op_rights |
info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1126/sciadv.adi0963 |
container_title |
Science Advances |
container_volume |
9 |
container_issue |
26 |
_version_ |
1786163354502430720 |