Characterization and Reshaping of a Large and Hydrophobic Nucleophile Pocket in Lipases/Acyltransferases
Abstract Lipases/acyltransferases, such as CpLIP2 from Candida parapsilosis and CduLAc from Candida dubliniensis , catalyze acyl transfer preferentially over hydrolysis if a suitable nucleophile is present, even in a medium with a high thermodynamic activity of water ( a W ). These enzymes are relat...
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crwiley:10.1002/cbic.201800279 2024-06-02T07:58:00+00:00 Characterization and Reshaping of a Large and Hydrophobic Nucleophile Pocket in Lipases/Acyltransferases Jan Deniau, Anne‐Hélène Subileau, Maeva Dubreucq, Eric 2018 http://dx.doi.org/10.1002/cbic.201800279 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fcbic.201800279 https://onlinelibrary.wiley.com/doi/pdf/10.1002/cbic.201800279 https://onlinelibrary.wiley.com/doi/full-xml/10.1002/cbic.201800279 en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor ChemBioChem volume 19, issue 17, page 1839-1844 ISSN 1439-4227 1439-7633 journal-article 2018 crwiley https://doi.org/10.1002/cbic.201800279 2024-05-03T11:47:12Z Abstract Lipases/acyltransferases, such as CpLIP2 from Candida parapsilosis and CduLAc from Candida dubliniensis , catalyze acyl transfer preferentially over hydrolysis if a suitable nucleophile is present, even in a medium with a high thermodynamic activity of water ( a W ). These enzymes are related to CAL‐A from Moesziomyces antarcticus , which, in comparison, displays a lower acyl transfer ability. The 3D structures of wild types and mutants of CAL‐A, CpLIP2, and CduLAc revealed differences in size and hydrophobicity of a large pocket located under the catalytic triad. The kinetic behavior of site‐directed mutants confirmed the role of this pocket in competition between methanol and water as the nucleophile acceptor for the deacylation step. The mutations provided a better understanding of key structural determinants for variable levels of acyltransferase ability observed and supported the existence of a complex network of nucleophile interactions within the enzymes. The shape and size of the possible nucleophile pocket identified also suggested that multiple binding sites could exist, which supported the hypothesis of non‐overlapping leaving and accepting nucleophile binding sites. Article in Journal/Newspaper Antarc* antarcticus Wiley Online Library ChemBioChem 19 17 1839 1844 |
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Wiley Online Library |
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crwiley |
language |
English |
description |
Abstract Lipases/acyltransferases, such as CpLIP2 from Candida parapsilosis and CduLAc from Candida dubliniensis , catalyze acyl transfer preferentially over hydrolysis if a suitable nucleophile is present, even in a medium with a high thermodynamic activity of water ( a W ). These enzymes are related to CAL‐A from Moesziomyces antarcticus , which, in comparison, displays a lower acyl transfer ability. The 3D structures of wild types and mutants of CAL‐A, CpLIP2, and CduLAc revealed differences in size and hydrophobicity of a large pocket located under the catalytic triad. The kinetic behavior of site‐directed mutants confirmed the role of this pocket in competition between methanol and water as the nucleophile acceptor for the deacylation step. The mutations provided a better understanding of key structural determinants for variable levels of acyltransferase ability observed and supported the existence of a complex network of nucleophile interactions within the enzymes. The shape and size of the possible nucleophile pocket identified also suggested that multiple binding sites could exist, which supported the hypothesis of non‐overlapping leaving and accepting nucleophile binding sites. |
format |
Article in Journal/Newspaper |
author |
Jan Deniau, Anne‐Hélène Subileau, Maeva Dubreucq, Eric |
spellingShingle |
Jan Deniau, Anne‐Hélène Subileau, Maeva Dubreucq, Eric Characterization and Reshaping of a Large and Hydrophobic Nucleophile Pocket in Lipases/Acyltransferases |
author_facet |
Jan Deniau, Anne‐Hélène Subileau, Maeva Dubreucq, Eric |
author_sort |
Jan Deniau, Anne‐Hélène |
title |
Characterization and Reshaping of a Large and Hydrophobic Nucleophile Pocket in Lipases/Acyltransferases |
title_short |
Characterization and Reshaping of a Large and Hydrophobic Nucleophile Pocket in Lipases/Acyltransferases |
title_full |
Characterization and Reshaping of a Large and Hydrophobic Nucleophile Pocket in Lipases/Acyltransferases |
title_fullStr |
Characterization and Reshaping of a Large and Hydrophobic Nucleophile Pocket in Lipases/Acyltransferases |
title_full_unstemmed |
Characterization and Reshaping of a Large and Hydrophobic Nucleophile Pocket in Lipases/Acyltransferases |
title_sort |
characterization and reshaping of a large and hydrophobic nucleophile pocket in lipases/acyltransferases |
publisher |
Wiley |
publishDate |
2018 |
url |
http://dx.doi.org/10.1002/cbic.201800279 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fcbic.201800279 https://onlinelibrary.wiley.com/doi/pdf/10.1002/cbic.201800279 https://onlinelibrary.wiley.com/doi/full-xml/10.1002/cbic.201800279 |
genre |
Antarc* antarcticus |
genre_facet |
Antarc* antarcticus |
op_source |
ChemBioChem volume 19, issue 17, page 1839-1844 ISSN 1439-4227 1439-7633 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1002/cbic.201800279 |
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ChemBioChem |
container_volume |
19 |
container_issue |
17 |
container_start_page |
1839 |
op_container_end_page |
1844 |
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1800741241273450496 |