A combined experimental and computational approach for the rationalization of the catalytic activity of lipase B from Candida antarctica in water–organic solvent mixtures
Abstract Background The addition of organic solvents to an aqueous medium for enzymatic reactions offers several advantages, as they can increase the solubility of substrates but can also lead to enzyme inactivation and/or aggregation. Results The effect of adding 30% of several water‐soluble organi...
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Online Access: | http://dx.doi.org/10.1002/jctb.7467 https://onlinelibrary.wiley.com/doi/pdf/10.1002/jctb.7467 |
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crwiley:10.1002/jctb.7467 2024-06-02T07:57:33+00:00 A combined experimental and computational approach for the rationalization of the catalytic activity of lipase B from Candida antarctica in water–organic solvent mixtures Vetrano, Alice Daidone, Isabella Spreti, Nicoletta Capone, Matteo 2023 http://dx.doi.org/10.1002/jctb.7467 https://onlinelibrary.wiley.com/doi/pdf/10.1002/jctb.7467 en eng Wiley http://creativecommons.org/licenses/by/4.0/ Journal of Chemical Technology & Biotechnology volume 98, issue 10, page 2429-2436 ISSN 0268-2575 1097-4660 journal-article 2023 crwiley https://doi.org/10.1002/jctb.7467 2024-05-03T11:27:03Z Abstract Background The addition of organic solvents to an aqueous medium for enzymatic reactions offers several advantages, as they can increase the solubility of substrates but can also lead to enzyme inactivation and/or aggregation. Results The effect of adding 30% of several water‐soluble organic solvents on the catalytic activity of lipase B from Candida antarctica (CalB) was studied and the results showed that the highest activity was obtained with the addition of t ‐butanol. t ‐Butanol and acetonitrile were selected and the kinetic parameters, determined to deepen their effect on CalB activity, showed that the addition of acetonitrile improved the enzyme–substrate affinity, while water– t ‐butanol mixtures led to a more than ninefold increase in k cat . To rationalize at a molecular level the kinetic results, molecular dynamic simulations were performed. Analysis of the accessibility of the active‐site cavity, solvent occupancy in the site and in the oxyanion hole, and the stability of the catalytic triad in the two solvent mixtures, provided insight into their effects on the catalytic properties of CalB. Conclusion The lower occupancy in the oxyanion hole of water molecules and a shorter residence time in the active site of acetonitrile molecules in the acetonitrile–water mixture contribute to the higher enzyme–substrate affinity found experimentally. Conversely, the higher k cat in the t ‐butanol mixture is explained by the higher stability of the catalytic triad and by an increase in the nucleophilicity of the catalytic serine due to the persistent presence of t ‐butanol molecules in the active site. © 2023 The Authors. Journal of Chemical Technology and Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry (SCI). Article in Journal/Newspaper Antarc* Antarctica Wiley Online Library Journal of Chemical Technology & Biotechnology 98 10 2429 2436 |
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English |
description |
Abstract Background The addition of organic solvents to an aqueous medium for enzymatic reactions offers several advantages, as they can increase the solubility of substrates but can also lead to enzyme inactivation and/or aggregation. Results The effect of adding 30% of several water‐soluble organic solvents on the catalytic activity of lipase B from Candida antarctica (CalB) was studied and the results showed that the highest activity was obtained with the addition of t ‐butanol. t ‐Butanol and acetonitrile were selected and the kinetic parameters, determined to deepen their effect on CalB activity, showed that the addition of acetonitrile improved the enzyme–substrate affinity, while water– t ‐butanol mixtures led to a more than ninefold increase in k cat . To rationalize at a molecular level the kinetic results, molecular dynamic simulations were performed. Analysis of the accessibility of the active‐site cavity, solvent occupancy in the site and in the oxyanion hole, and the stability of the catalytic triad in the two solvent mixtures, provided insight into their effects on the catalytic properties of CalB. Conclusion The lower occupancy in the oxyanion hole of water molecules and a shorter residence time in the active site of acetonitrile molecules in the acetonitrile–water mixture contribute to the higher enzyme–substrate affinity found experimentally. Conversely, the higher k cat in the t ‐butanol mixture is explained by the higher stability of the catalytic triad and by an increase in the nucleophilicity of the catalytic serine due to the persistent presence of t ‐butanol molecules in the active site. © 2023 The Authors. Journal of Chemical Technology and Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry (SCI). |
format |
Article in Journal/Newspaper |
author |
Vetrano, Alice Daidone, Isabella Spreti, Nicoletta Capone, Matteo |
spellingShingle |
Vetrano, Alice Daidone, Isabella Spreti, Nicoletta Capone, Matteo A combined experimental and computational approach for the rationalization of the catalytic activity of lipase B from Candida antarctica in water–organic solvent mixtures |
author_facet |
Vetrano, Alice Daidone, Isabella Spreti, Nicoletta Capone, Matteo |
author_sort |
Vetrano, Alice |
title |
A combined experimental and computational approach for the rationalization of the catalytic activity of lipase B from Candida antarctica in water–organic solvent mixtures |
title_short |
A combined experimental and computational approach for the rationalization of the catalytic activity of lipase B from Candida antarctica in water–organic solvent mixtures |
title_full |
A combined experimental and computational approach for the rationalization of the catalytic activity of lipase B from Candida antarctica in water–organic solvent mixtures |
title_fullStr |
A combined experimental and computational approach for the rationalization of the catalytic activity of lipase B from Candida antarctica in water–organic solvent mixtures |
title_full_unstemmed |
A combined experimental and computational approach for the rationalization of the catalytic activity of lipase B from Candida antarctica in water–organic solvent mixtures |
title_sort |
combined experimental and computational approach for the rationalization of the catalytic activity of lipase b from candida antarctica in water–organic solvent mixtures |
publisher |
Wiley |
publishDate |
2023 |
url |
http://dx.doi.org/10.1002/jctb.7467 https://onlinelibrary.wiley.com/doi/pdf/10.1002/jctb.7467 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Journal of Chemical Technology & Biotechnology volume 98, issue 10, page 2429-2436 ISSN 0268-2575 1097-4660 |
op_rights |
http://creativecommons.org/licenses/by/4.0/ |
op_doi |
https://doi.org/10.1002/jctb.7467 |
container_title |
Journal of Chemical Technology & Biotechnology |
container_volume |
98 |
container_issue |
10 |
container_start_page |
2429 |
op_container_end_page |
2436 |
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1800740715634884608 |