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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Published in:Science Advances
Main Authors: van der Ent, Florian, Skagseth, Susann, Lund, Bjarte A., Socan, Jaka, Griese, Julia J., Brandsdal, Bjorn O., Åqvist, Johan
Format: Article in Journal/Newspaper
Language:English
Published: Uppsala universitet, Beräkningsbiologi och bioinformatik 2023
Subjects:
Online Access:http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-508544
https://doi.org/10.1126/sciadv.adi0963
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spelling 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
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