Comparison of Candida antarctica Lipase B Variants for Conversion of ε-Caprolactone in Aqueous Medium-Part 2
Enzyme-catalyzed ring-opening polymerization of lactones is a method of increasing interest for the synthesis of polyesters. In the present work, we investigated which changes in the structure of Candida antarctica lipase B (CaLB) shift the catalytic equilibrium between esterification and hydrolysis...
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ftleibnizopen:oai:oai.leibnizopen.de:jnbgXIkBdbrxVwz6fiNy 2023-07-30T03:58:12+02:00 Comparison of Candida antarctica Lipase B Variants for Conversion of ε-Caprolactone in Aqueous Medium-Part 2 Höck, Heidi Engel, Stefan Weingarten, Simone Keul, Helmut Schwaneberg, Ulrich Möller, Martin Bocola, Marco 2018 application/pdf https://oa.tib.eu/renate/handle/123456789/8646 https://doi.org/10.34657/7684 eng eng Basel : MDPI CC BY 4.0 Unported https://creativecommons.org/licenses/by/4.0/ Polymers 10 (2018), Nr. 5 Candida antarctica lipase B Enzymatic ring-opening polymerization Enzyme engineering Immobilization Microgel 540 article Text 2018 ftleibnizopen https://doi.org/10.34657/7684 2023-07-16T23:22:24Z Enzyme-catalyzed ring-opening polymerization of lactones is a method of increasing interest for the synthesis of polyesters. In the present work, we investigated which changes in the structure of Candida antarctica lipase B (CaLB) shift the catalytic equilibrium between esterification and hydrolysis towards polymerization. Therefore, we present two concepts: (i) removing the glycosylation of CaLB to increase the surface hydrophobicity; and (ii) introducing a hydrophobic lid adapted from Pseudomonas cepacia lipase (PsCL) to enhance the interaction of a growing polymer chain to the elongated lid helix. The deglycosylated CaLB (CaLB-degl) was successfully generated by site-saturation mutagenesis of asparagine 74. Furthermore, computational modeling showed that the introduction of a lid helix at position Ala148 was structurally feasible and the geometry of the active site remained intact. Via overlap extension PCR the lid was successfully inserted, and the variant was produced in large scale in Pichia pastoris with glycosylation (CaLB-lid) and without (CaLB-degl-lid). While the lid variants show a minor positive effect on the polymerization activity, CaLB-degl showed a clearly reduced hydrolytic and enhanced polymerization activity. Immobilization in a hydrophobic polyglycidol-based microgel intensified this effect such that a higher polymerization activity was achieved, compared to the “gold standard” Novozym® 435. publishedVersion Article in Journal/Newspaper Antarc* Antarctica LeibnizOpen (The Leibniz Association) |
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Open Polar |
collection |
LeibnizOpen (The Leibniz Association) |
op_collection_id |
ftleibnizopen |
language |
English |
topic |
Candida antarctica lipase B Enzymatic ring-opening polymerization Enzyme engineering Immobilization Microgel 540 |
spellingShingle |
Candida antarctica lipase B Enzymatic ring-opening polymerization Enzyme engineering Immobilization Microgel 540 Höck, Heidi Engel, Stefan Weingarten, Simone Keul, Helmut Schwaneberg, Ulrich Möller, Martin Bocola, Marco Comparison of Candida antarctica Lipase B Variants for Conversion of ε-Caprolactone in Aqueous Medium-Part 2 |
topic_facet |
Candida antarctica lipase B Enzymatic ring-opening polymerization Enzyme engineering Immobilization Microgel 540 |
description |
Enzyme-catalyzed ring-opening polymerization of lactones is a method of increasing interest for the synthesis of polyesters. In the present work, we investigated which changes in the structure of Candida antarctica lipase B (CaLB) shift the catalytic equilibrium between esterification and hydrolysis towards polymerization. Therefore, we present two concepts: (i) removing the glycosylation of CaLB to increase the surface hydrophobicity; and (ii) introducing a hydrophobic lid adapted from Pseudomonas cepacia lipase (PsCL) to enhance the interaction of a growing polymer chain to the elongated lid helix. The deglycosylated CaLB (CaLB-degl) was successfully generated by site-saturation mutagenesis of asparagine 74. Furthermore, computational modeling showed that the introduction of a lid helix at position Ala148 was structurally feasible and the geometry of the active site remained intact. Via overlap extension PCR the lid was successfully inserted, and the variant was produced in large scale in Pichia pastoris with glycosylation (CaLB-lid) and without (CaLB-degl-lid). While the lid variants show a minor positive effect on the polymerization activity, CaLB-degl showed a clearly reduced hydrolytic and enhanced polymerization activity. Immobilization in a hydrophobic polyglycidol-based microgel intensified this effect such that a higher polymerization activity was achieved, compared to the “gold standard” Novozym® 435. publishedVersion |
format |
Article in Journal/Newspaper |
author |
Höck, Heidi Engel, Stefan Weingarten, Simone Keul, Helmut Schwaneberg, Ulrich Möller, Martin Bocola, Marco |
author_facet |
Höck, Heidi Engel, Stefan Weingarten, Simone Keul, Helmut Schwaneberg, Ulrich Möller, Martin Bocola, Marco |
author_sort |
Höck, Heidi |
title |
Comparison of Candida antarctica Lipase B Variants for Conversion of ε-Caprolactone in Aqueous Medium-Part 2 |
title_short |
Comparison of Candida antarctica Lipase B Variants for Conversion of ε-Caprolactone in Aqueous Medium-Part 2 |
title_full |
Comparison of Candida antarctica Lipase B Variants for Conversion of ε-Caprolactone in Aqueous Medium-Part 2 |
title_fullStr |
Comparison of Candida antarctica Lipase B Variants for Conversion of ε-Caprolactone in Aqueous Medium-Part 2 |
title_full_unstemmed |
Comparison of Candida antarctica Lipase B Variants for Conversion of ε-Caprolactone in Aqueous Medium-Part 2 |
title_sort |
comparison of candida antarctica lipase b variants for conversion of ε-caprolactone in aqueous medium-part 2 |
publisher |
Basel : MDPI |
publishDate |
2018 |
url |
https://oa.tib.eu/renate/handle/123456789/8646 https://doi.org/10.34657/7684 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Polymers 10 (2018), Nr. 5 |
op_rights |
CC BY 4.0 Unported https://creativecommons.org/licenses/by/4.0/ |
op_doi |
https://doi.org/10.34657/7684 |
_version_ |
1772821073914494976 |