Contribution of both catalytic constant and Michaelis constant to CALB enantioselectivity; use of FEP calculations for prediction studies
International audience Candida antarctica lipase B (CALB) is characterised by its stability and ease of production and is widely used in the pharmaceutical industry. Here we report on the enantioselectivity of the enzyme using both experimental and computational methods. The apparent kinetic paramet...
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ftccsdartic:oai:HAL:hal-00789569v1 2023-05-15T14:04:01+02:00 Contribution of both catalytic constant and Michaelis constant to CALB enantioselectivity; use of FEP calculations for prediction studies Chaput, L. Sanejouand, Yves-Henri Balloumi, Asma Tran, Vinh Graber, Marianne LIttoral ENvironnement et Sociétés - UMRi 7266 (LIENSs) Université de La Rochelle (ULR)-Centre National de la Recherche Scientifique (CNRS) Unité de Biotechnologie, Biocatalyse et Biorégulation (U3B) Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS) 2012-02-24 https://hal.archives-ouvertes.fr/hal-00789569 https://hal.archives-ouvertes.fr/hal-00789569/document https://hal.archives-ouvertes.fr/hal-00789569/file/elsarticle_v7.pdf en eng HAL CCSD Elsevier hal-00789569 https://hal.archives-ouvertes.fr/hal-00789569 https://hal.archives-ouvertes.fr/hal-00789569/document https://hal.archives-ouvertes.fr/hal-00789569/file/elsarticle_v7.pdf info:eu-repo/semantics/OpenAccess ISSN: 1381-1177 Journal of Molecular Catalysis B: Enzymatic https://hal.archives-ouvertes.fr/hal-00789569 Journal of Molecular Catalysis B: Enzymatic, Elsevier, 2012, 76, pp.29-36 [SDV.BIO]Life Sciences [q-bio]/Biotechnology [INFO.INFO-BT]Computer Science [cs]/Biotechnology info:eu-repo/semantics/article Journal articles 2012 ftccsdartic 2021-10-24T15:24:55Z International audience Candida antarctica lipase B (CALB) is characterised by its stability and ease of production and is widely used in the pharmaceutical industry. Here we report on the enantioselectivity of the enzyme using both experimental and computational methods. The apparent kinetic parameters were first experimentally determined for enantiopure butan-2-ol and pentan-2-ol substrates. We demon- strate that enantiopreference for the R form of butan-2-ol arises mainly from a lower apparent K M . This corresponds to a major contribution of ΔΔG ES , the free energy difference between the ES complex formed with the R and S enantiomers, to ΔΔG‡, the free energy difference between both transition states, in comparison with ΔΔG kcat , the activation free energy difference. In the case of pentan-2-ol, we show that the enantiopreference for the R form comes from both a lower K M and a higher k cat . In addition, we used, for the first time, the Free Energy Perturbation method to evaluate the free energy difference between tetrahedral intermediates formed with R and S alcohol enantiomers for a series of secondary alcohols . This is a valid model for ΔΔG‡ Computational results were found to be in good agreement with experimental data, and enable the determination of substrate orientation in the active site with fair confidence. Article in Journal/Newspaper Antarc* Antarctica Archive ouverte HAL (Hyper Article en Ligne, CCSD - Centre pour la Communication Scientifique Directe) |
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Archive ouverte HAL (Hyper Article en Ligne, CCSD - Centre pour la Communication Scientifique Directe) |
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ftccsdartic |
language |
English |
topic |
[SDV.BIO]Life Sciences [q-bio]/Biotechnology [INFO.INFO-BT]Computer Science [cs]/Biotechnology |
spellingShingle |
[SDV.BIO]Life Sciences [q-bio]/Biotechnology [INFO.INFO-BT]Computer Science [cs]/Biotechnology Chaput, L. Sanejouand, Yves-Henri Balloumi, Asma Tran, Vinh Graber, Marianne Contribution of both catalytic constant and Michaelis constant to CALB enantioselectivity; use of FEP calculations for prediction studies |
topic_facet |
[SDV.BIO]Life Sciences [q-bio]/Biotechnology [INFO.INFO-BT]Computer Science [cs]/Biotechnology |
description |
International audience Candida antarctica lipase B (CALB) is characterised by its stability and ease of production and is widely used in the pharmaceutical industry. Here we report on the enantioselectivity of the enzyme using both experimental and computational methods. The apparent kinetic parameters were first experimentally determined for enantiopure butan-2-ol and pentan-2-ol substrates. We demon- strate that enantiopreference for the R form of butan-2-ol arises mainly from a lower apparent K M . This corresponds to a major contribution of ΔΔG ES , the free energy difference between the ES complex formed with the R and S enantiomers, to ΔΔG‡, the free energy difference between both transition states, in comparison with ΔΔG kcat , the activation free energy difference. In the case of pentan-2-ol, we show that the enantiopreference for the R form comes from both a lower K M and a higher k cat . In addition, we used, for the first time, the Free Energy Perturbation method to evaluate the free energy difference between tetrahedral intermediates formed with R and S alcohol enantiomers for a series of secondary alcohols . This is a valid model for ΔΔG‡ Computational results were found to be in good agreement with experimental data, and enable the determination of substrate orientation in the active site with fair confidence. |
author2 |
LIttoral ENvironnement et Sociétés - UMRi 7266 (LIENSs) Université de La Rochelle (ULR)-Centre National de la Recherche Scientifique (CNRS) Unité de Biotechnologie, Biocatalyse et Biorégulation (U3B) Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS) |
format |
Article in Journal/Newspaper |
author |
Chaput, L. Sanejouand, Yves-Henri Balloumi, Asma Tran, Vinh Graber, Marianne |
author_facet |
Chaput, L. Sanejouand, Yves-Henri Balloumi, Asma Tran, Vinh Graber, Marianne |
author_sort |
Chaput, L. |
title |
Contribution of both catalytic constant and Michaelis constant to CALB enantioselectivity; use of FEP calculations for prediction studies |
title_short |
Contribution of both catalytic constant and Michaelis constant to CALB enantioselectivity; use of FEP calculations for prediction studies |
title_full |
Contribution of both catalytic constant and Michaelis constant to CALB enantioselectivity; use of FEP calculations for prediction studies |
title_fullStr |
Contribution of both catalytic constant and Michaelis constant to CALB enantioselectivity; use of FEP calculations for prediction studies |
title_full_unstemmed |
Contribution of both catalytic constant and Michaelis constant to CALB enantioselectivity; use of FEP calculations for prediction studies |
title_sort |
contribution of both catalytic constant and michaelis constant to calb enantioselectivity; use of fep calculations for prediction studies |
publisher |
HAL CCSD |
publishDate |
2012 |
url |
https://hal.archives-ouvertes.fr/hal-00789569 https://hal.archives-ouvertes.fr/hal-00789569/document https://hal.archives-ouvertes.fr/hal-00789569/file/elsarticle_v7.pdf |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
ISSN: 1381-1177 Journal of Molecular Catalysis B: Enzymatic https://hal.archives-ouvertes.fr/hal-00789569 Journal of Molecular Catalysis B: Enzymatic, Elsevier, 2012, 76, pp.29-36 |
op_relation |
hal-00789569 https://hal.archives-ouvertes.fr/hal-00789569 https://hal.archives-ouvertes.fr/hal-00789569/document https://hal.archives-ouvertes.fr/hal-00789569/file/elsarticle_v7.pdf |
op_rights |
info:eu-repo/semantics/OpenAccess |
_version_ |
1766274955151933440 |