Interfacial activation of Candida antarctica lipase B: combined evidence from experiment and simulation
Lipase immobilization is frequently used for altering the catalytic properties of these industrially used enzymes. Many lipases bind strongly to hydrophobic surfaces where they undergo interfacial activation. Candida antarctica lipase B (CalB), one of the most commonly used biocatalysts, is frequent...
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ftunivqespace:oai:espace.library.uq.edu.au:UQ:597255 2023-05-15T13:47:31+02:00 Interfacial activation of Candida antarctica lipase B: combined evidence from experiment and simulation Zisis, Themistoklis Freddolino, Peter L. Turunen, Petri van Teeseing, Muriel C. F. Rowan, Alan E. Blank, Kerstin G. 2015-09-29 https://espace.library.uq.edu.au/view/UQ:597255 eng eng American Chemical Society doi:10.1021/acs.biochem.5b00586 issn:1520-4995 issn:0006-2960 orcid:0000-0003-0728-3851 2009-10016B 700.58.430 024.001.035 K99/R00GM097033 TG-MCA05S028 R00GM097033 K99GM097033 Biochemistry & Molecular Biology 1303 Biochemistry Journal Article 2015 ftunivqespace https://doi.org/10.1021/acs.biochem.5b00586 2020-11-24T02:12:25Z Lipase immobilization is frequently used for altering the catalytic properties of these industrially used enzymes. Many lipases bind strongly to hydrophobic surfaces where they undergo interfacial activation. Candida antarctica lipase B (CalB), one of the most commonly used biocatalysts, is frequently discussed as an atypical lipase lacking interfacial activation. Here we show that CalB displays an enhanced catalytic rate for large, bulky substrates when adsorbed to a hydrophobic interface composed of densely packed alkyl chains. We attribute this increased activity of more than 7-fold to a conformational change that yields a more open active site. This hypothesis is supported by molecular dynamics simulations that show a high mobility for a small "lid" (helix α5) close to the active site. Molecular docking calculations confirm that a highly open conformation of this helix is required for binding large, bulky substrates and that this conformation is favored in a hydrophobic environment. Taken together, our combined approach provides clear evidence for the interfacial activation of CalB on highly hydrophobic surfaces. In contrast to other lipases, however, the conformational change only affects large, bulky substrates, leading to the conclusion that CalB acts like an esterase for small substrates and as a lipase for substrates with large alcohol substituents. Article in Journal/Newspaper Antarc* Antarctica The University of Queensland: UQ eSpace Biochemistry 54 38 5969 5979 |
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Open Polar |
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The University of Queensland: UQ eSpace |
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ftunivqespace |
language |
English |
topic |
Biochemistry & Molecular Biology 1303 Biochemistry |
spellingShingle |
Biochemistry & Molecular Biology 1303 Biochemistry Zisis, Themistoklis Freddolino, Peter L. Turunen, Petri van Teeseing, Muriel C. F. Rowan, Alan E. Blank, Kerstin G. Interfacial activation of Candida antarctica lipase B: combined evidence from experiment and simulation |
topic_facet |
Biochemistry & Molecular Biology 1303 Biochemistry |
description |
Lipase immobilization is frequently used for altering the catalytic properties of these industrially used enzymes. Many lipases bind strongly to hydrophobic surfaces where they undergo interfacial activation. Candida antarctica lipase B (CalB), one of the most commonly used biocatalysts, is frequently discussed as an atypical lipase lacking interfacial activation. Here we show that CalB displays an enhanced catalytic rate for large, bulky substrates when adsorbed to a hydrophobic interface composed of densely packed alkyl chains. We attribute this increased activity of more than 7-fold to a conformational change that yields a more open active site. This hypothesis is supported by molecular dynamics simulations that show a high mobility for a small "lid" (helix α5) close to the active site. Molecular docking calculations confirm that a highly open conformation of this helix is required for binding large, bulky substrates and that this conformation is favored in a hydrophobic environment. Taken together, our combined approach provides clear evidence for the interfacial activation of CalB on highly hydrophobic surfaces. In contrast to other lipases, however, the conformational change only affects large, bulky substrates, leading to the conclusion that CalB acts like an esterase for small substrates and as a lipase for substrates with large alcohol substituents. |
format |
Article in Journal/Newspaper |
author |
Zisis, Themistoklis Freddolino, Peter L. Turunen, Petri van Teeseing, Muriel C. F. Rowan, Alan E. Blank, Kerstin G. |
author_facet |
Zisis, Themistoklis Freddolino, Peter L. Turunen, Petri van Teeseing, Muriel C. F. Rowan, Alan E. Blank, Kerstin G. |
author_sort |
Zisis, Themistoklis |
title |
Interfacial activation of Candida antarctica lipase B: combined evidence from experiment and simulation |
title_short |
Interfacial activation of Candida antarctica lipase B: combined evidence from experiment and simulation |
title_full |
Interfacial activation of Candida antarctica lipase B: combined evidence from experiment and simulation |
title_fullStr |
Interfacial activation of Candida antarctica lipase B: combined evidence from experiment and simulation |
title_full_unstemmed |
Interfacial activation of Candida antarctica lipase B: combined evidence from experiment and simulation |
title_sort |
interfacial activation of candida antarctica lipase b: combined evidence from experiment and simulation |
publisher |
American Chemical Society |
publishDate |
2015 |
url |
https://espace.library.uq.edu.au/view/UQ:597255 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
doi:10.1021/acs.biochem.5b00586 issn:1520-4995 issn:0006-2960 orcid:0000-0003-0728-3851 2009-10016B 700.58.430 024.001.035 K99/R00GM097033 TG-MCA05S028 R00GM097033 K99GM097033 |
op_doi |
https://doi.org/10.1021/acs.biochem.5b00586 |
container_title |
Biochemistry |
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54 |
container_issue |
38 |
container_start_page |
5969 |
op_container_end_page |
5979 |
_version_ |
1766247258564591616 |