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...
Published in: | Biochemistry |
---|---|
Main Authors: | , , , , , |
Format: | Text |
Language: | English |
Published: |
2015
|
Subjects: | |
Online Access: | http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4697882/ http://www.ncbi.nlm.nih.gov/pubmed/26346632 https://doi.org/10.1021/acs.biochem.5b00586 |
id |
ftpubmed:oai:pubmedcentral.nih.gov:4697882 |
---|---|
record_format |
openpolar |
spelling |
ftpubmed:oai:pubmedcentral.nih.gov:4697882 2023-05-15T13:36:46+02:00 Interfacial activation of Candida antarctica lipase B: combined evidence from experiment and simulation Zisis, Themistoklis Freddolino, Peter L. Turunen, Petri van Teeseling, Muriel C. F. Rowan, Alan E. Blank, Kerstin G. 2015-09-15 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4697882/ http://www.ncbi.nlm.nih.gov/pubmed/26346632 https://doi.org/10.1021/acs.biochem.5b00586 en eng http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4697882/ http://www.ncbi.nlm.nih.gov/pubmed/26346632 http://dx.doi.org/10.1021/acs.biochem.5b00586 Article Text 2015 ftpubmed https://doi.org/10.1021/acs.biochem.5b00586 2016-01-03T01:39:22Z 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. Text Antarc* Antarctica PubMed Central (PMC) Biochemistry 54 38 5969 5979 |
institution |
Open Polar |
collection |
PubMed Central (PMC) |
op_collection_id |
ftpubmed |
language |
English |
topic |
Article |
spellingShingle |
Article Zisis, Themistoklis Freddolino, Peter L. Turunen, Petri van Teeseling, Muriel C. F. Rowan, Alan E. Blank, Kerstin G. Interfacial activation of Candida antarctica lipase B: combined evidence from experiment and simulation |
topic_facet |
Article |
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 |
Text |
author |
Zisis, Themistoklis Freddolino, Peter L. Turunen, Petri van Teeseling, Muriel C. F. Rowan, Alan E. Blank, Kerstin G. |
author_facet |
Zisis, Themistoklis Freddolino, Peter L. Turunen, Petri van Teeseling, 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 |
publishDate |
2015 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4697882/ http://www.ncbi.nlm.nih.gov/pubmed/26346632 https://doi.org/10.1021/acs.biochem.5b00586 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4697882/ http://www.ncbi.nlm.nih.gov/pubmed/26346632 http://dx.doi.org/10.1021/acs.biochem.5b00586 |
op_doi |
https://doi.org/10.1021/acs.biochem.5b00586 |
container_title |
Biochemistry |
container_volume |
54 |
container_issue |
38 |
container_start_page |
5969 |
op_container_end_page |
5979 |
_version_ |
1766083568540319744 |