Organic-Solvent-Tolerant Carboxylic Ester Hydrolases for Organic Synthesis
Biocatalysis has emerged as an important tool in synthetic organic chemistry enabling the chemical industry to execute reactions with high regio- or enantioselectivity and under usually mild reaction conditions while avoiding toxic waste. Target substrates and products of reactions catalyzed by carb...
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ftfzjuelichnvdb:oai:juser.fz-juelich.de:888809 2023-05-15T13:35:13+02:00 Organic-Solvent-Tolerant Carboxylic Ester Hydrolases for Organic Synthesis Bollinger, Alexander Molitor, Rebecka Thies, Stephan Koch, Rainhard Coscolín, Cristina Ferrer, Manuel Jaeger, Karl-Erich DE 2020 https://juser.fz-juelich.de/record/888809 https://juser.fz-juelich.de/search?p=id:%22FZJ-2020-05223%22 eng eng Soc. info:eu-repo/semantics/altIdentifier/doi/10.1128/AEM.00106-20 info:eu-repo/semantics/altIdentifier/issn/1070-6291 info:eu-repo/semantics/altIdentifier/issn/0099-2240 info:eu-repo/semantics/altIdentifier/wos/WOS:000529303800005 info:eu-repo/semantics/altIdentifier/pmid/32111588 info:eu-repo/semantics/altIdentifier/issn/1098-5336 info:eu-repo/semantics/altIdentifier/hdl/2128/26517 info:eu-repo/semantics/altIdentifier/issn/0003-6919 https://juser.fz-juelich.de/record/888809 https://juser.fz-juelich.de/search?p=id:%22FZJ-2020-05223%22 info:eu-repo/semantics/openAccess Applied and environmental microbiology 86(9), e00106-20 (2020). doi:10.1128/AEM.00106-20 info:eu-repo/classification/ddc/570 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 2020 ftfzjuelichnvdb https://doi.org/10.1128/AEM.00106-20 2022-07-14T11:21:25Z Biocatalysis has emerged as an important tool in synthetic organic chemistry enabling the chemical industry to execute reactions with high regio- or enantioselectivity and under usually mild reaction conditions while avoiding toxic waste. Target substrates and products of reactions catalyzed by carboxylic ester hydrolases are often poorly water soluble and require organic solvents, whereas enzymes are evolved by nature to be active in cells, i.e., in aqueous rather than organic solvents. Therefore, biocatalysts that withstand organic solvents are urgently needed. Current strategies to identify such enzymes rely on laborious tests carried out by incubation in different organic solvents and determination of residual activity. Here, we describe a simple assay useful for screening large libraries of carboxylic ester hydrolases for resistance and activity in water-miscible organic solvents. We have screened a set of 26 enzymes, most of them identified in this study, with four different water-miscible organic solvents. The triglyceride tributyrin was used as a substrate, and fatty acids released by enzymatic hydrolysis were detected by a pH shift indicated by the indicator dye nitrazine yellow. With this strategy, we succeeded in identifying a novel highly organic-solvent-tolerant esterase from Pseudomonas aestusnigri. In addition, the newly identified enzymes were tested with sterically demanding substrates, which are common in pharmaceutical intermediates, and two enzymes from Alcanivorax borkumensis were identified which outcompeted the gold standard ester hydrolase CalB from Candida antarctica Article in Journal/Newspaper Antarc* Antarctica Forschungszentrum Jülich: JuSER (Juelich Shared Electronic Resources) Applied and Environmental Microbiology 86 9 |
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Forschungszentrum Jülich: JuSER (Juelich Shared Electronic Resources) |
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English |
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info:eu-repo/classification/ddc/570 |
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info:eu-repo/classification/ddc/570 Bollinger, Alexander Molitor, Rebecka Thies, Stephan Koch, Rainhard Coscolín, Cristina Ferrer, Manuel Jaeger, Karl-Erich Organic-Solvent-Tolerant Carboxylic Ester Hydrolases for Organic Synthesis |
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info:eu-repo/classification/ddc/570 |
description |
Biocatalysis has emerged as an important tool in synthetic organic chemistry enabling the chemical industry to execute reactions with high regio- or enantioselectivity and under usually mild reaction conditions while avoiding toxic waste. Target substrates and products of reactions catalyzed by carboxylic ester hydrolases are often poorly water soluble and require organic solvents, whereas enzymes are evolved by nature to be active in cells, i.e., in aqueous rather than organic solvents. Therefore, biocatalysts that withstand organic solvents are urgently needed. Current strategies to identify such enzymes rely on laborious tests carried out by incubation in different organic solvents and determination of residual activity. Here, we describe a simple assay useful for screening large libraries of carboxylic ester hydrolases for resistance and activity in water-miscible organic solvents. We have screened a set of 26 enzymes, most of them identified in this study, with four different water-miscible organic solvents. The triglyceride tributyrin was used as a substrate, and fatty acids released by enzymatic hydrolysis were detected by a pH shift indicated by the indicator dye nitrazine yellow. With this strategy, we succeeded in identifying a novel highly organic-solvent-tolerant esterase from Pseudomonas aestusnigri. In addition, the newly identified enzymes were tested with sterically demanding substrates, which are common in pharmaceutical intermediates, and two enzymes from Alcanivorax borkumensis were identified which outcompeted the gold standard ester hydrolase CalB from Candida antarctica |
format |
Article in Journal/Newspaper |
author |
Bollinger, Alexander Molitor, Rebecka Thies, Stephan Koch, Rainhard Coscolín, Cristina Ferrer, Manuel Jaeger, Karl-Erich |
author_facet |
Bollinger, Alexander Molitor, Rebecka Thies, Stephan Koch, Rainhard Coscolín, Cristina Ferrer, Manuel Jaeger, Karl-Erich |
author_sort |
Bollinger, Alexander |
title |
Organic-Solvent-Tolerant Carboxylic Ester Hydrolases for Organic Synthesis |
title_short |
Organic-Solvent-Tolerant Carboxylic Ester Hydrolases for Organic Synthesis |
title_full |
Organic-Solvent-Tolerant Carboxylic Ester Hydrolases for Organic Synthesis |
title_fullStr |
Organic-Solvent-Tolerant Carboxylic Ester Hydrolases for Organic Synthesis |
title_full_unstemmed |
Organic-Solvent-Tolerant Carboxylic Ester Hydrolases for Organic Synthesis |
title_sort |
organic-solvent-tolerant carboxylic ester hydrolases for organic synthesis |
publisher |
Soc. |
publishDate |
2020 |
url |
https://juser.fz-juelich.de/record/888809 https://juser.fz-juelich.de/search?p=id:%22FZJ-2020-05223%22 |
op_coverage |
DE |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Applied and environmental microbiology 86(9), e00106-20 (2020). doi:10.1128/AEM.00106-20 |
op_relation |
info:eu-repo/semantics/altIdentifier/doi/10.1128/AEM.00106-20 info:eu-repo/semantics/altIdentifier/issn/1070-6291 info:eu-repo/semantics/altIdentifier/issn/0099-2240 info:eu-repo/semantics/altIdentifier/wos/WOS:000529303800005 info:eu-repo/semantics/altIdentifier/pmid/32111588 info:eu-repo/semantics/altIdentifier/issn/1098-5336 info:eu-repo/semantics/altIdentifier/hdl/2128/26517 info:eu-repo/semantics/altIdentifier/issn/0003-6919 https://juser.fz-juelich.de/record/888809 https://juser.fz-juelich.de/search?p=id:%22FZJ-2020-05223%22 |
op_rights |
info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1128/AEM.00106-20 |
container_title |
Applied and Environmental Microbiology |
container_volume |
86 |
container_issue |
9 |
_version_ |
1766062974692229120 |