The Metagenome-Derived Enzymes LipS and LipT Increase the Diversity of Known Lipases
Triacylglycerol lipases (EC 3.1.1.3) catalyze both hydrolysis and synthesis reactions with a broad spectrum of substrates rendering them especially suitable for many biotechnological applications. Most lipases used today originate from mesophilic organisms and are susceptible to thermal denaturation...
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ftfzjuelichnvdb:oai:juser.fz-juelich.de:202186 2023-05-15T13:41:25+02:00 The Metagenome-Derived Enzymes LipS and LipT Increase the Diversity of Known Lipases Chow, Jennifer Kovacic, Filip Menyes, Ina Bornscheuer, Uwe T. Eckstein, Marrit Thum, Oliver Liese, Andreas Mueller-Dieckmann, Jochen Jaeger, Karl-Erich Streit, Wolfgang R. Dall Antonia, Yuliya Krauss, Ulrich Fersini, Francesco Schmeisser, Christel Lauinger, Benjamin Bongen, Patrick Pietruszka, Jörg Schmidt, Marlen DE 2012 https://juser.fz-juelich.de/record/202186 https://juser.fz-juelich.de/search?p=id:%22FZJ-2015-04475%22 eng eng PLoS info:eu-repo/semantics/altIdentifier/wos/WOS:000310310200064 info:eu-repo/semantics/altIdentifier/issn/1932-6203 info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0047665 info:eu-repo/semantics/altIdentifier/hdl/2128/8945 https://juser.fz-juelich.de/record/202186 https://juser.fz-juelich.de/search?p=id:%22FZJ-2015-04475%22 info:eu-repo/semantics/openAccess PLoS one 7(10), e47665 - (2012). doi:10.1371/journal.pone.0047665 info:eu-repo/classification/ddc/500 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 2012 ftfzjuelichnvdb https://doi.org/10.1371/journal.pone.0047665 2022-07-14T11:07:10Z Triacylglycerol lipases (EC 3.1.1.3) catalyze both hydrolysis and synthesis reactions with a broad spectrum of substrates rendering them especially suitable for many biotechnological applications. Most lipases used today originate from mesophilic organisms and are susceptible to thermal denaturation whereas only few possess high thermotolerance. Here, we report on the identification and characterization of two novel thermostable bacterial lipases identified by functional metagenomic screenings. Metagenomic libraries were constructed from enrichment cultures maintained at 65 to 75°C and screened resulting in the identification of initially 10 clones with lipolytic activities. Subsequently, two ORFs were identified encoding lipases, LipS and LipT. Comparative sequence analyses suggested that both enzymes are members of novel lipase families. LipS is a 30.2 kDa protein and revealed a half-life of 48 h at 70°C. The lipT gene encoded for a multimeric enzyme with a half-life of 3 h at 70°C. LipS had an optimum temperature at 70°C and LipT at 75°C. Both enzymes catalyzed hydrolysis of long-chain (C12 and C14) fatty acid esters and additionally hydrolyzed a number of industry-relevant substrates. LipS was highly specific for (R)-ibuprofen-phenyl ester with an enantiomeric excess (ee) of 99%. Furthermore, LipS was able to synthesize 1-propyl laurate and 1-tetradecyl myristate at 70°C with rates similar to those of the lipase CalB from Candida antarctica. LipS represents the first example of a thermostable metagenome-derived lipase with significant synthesis activities. Its X-ray structure was solved with a resolution of 1.99 Å revealing an unusually compact lid structure. Article in Journal/Newspaper Antarc* Antarctica Forschungszentrum Jülich: JuSER (Juelich Shared Electronic Resources) PLoS ONE 7 10 e47665 |
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Forschungszentrum Jülich: JuSER (Juelich Shared Electronic Resources) |
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language |
English |
topic |
info:eu-repo/classification/ddc/500 |
spellingShingle |
info:eu-repo/classification/ddc/500 Chow, Jennifer Kovacic, Filip Menyes, Ina Bornscheuer, Uwe T. Eckstein, Marrit Thum, Oliver Liese, Andreas Mueller-Dieckmann, Jochen Jaeger, Karl-Erich Streit, Wolfgang R. Dall Antonia, Yuliya Krauss, Ulrich Fersini, Francesco Schmeisser, Christel Lauinger, Benjamin Bongen, Patrick Pietruszka, Jörg Schmidt, Marlen The Metagenome-Derived Enzymes LipS and LipT Increase the Diversity of Known Lipases |
topic_facet |
info:eu-repo/classification/ddc/500 |
description |
Triacylglycerol lipases (EC 3.1.1.3) catalyze both hydrolysis and synthesis reactions with a broad spectrum of substrates rendering them especially suitable for many biotechnological applications. Most lipases used today originate from mesophilic organisms and are susceptible to thermal denaturation whereas only few possess high thermotolerance. Here, we report on the identification and characterization of two novel thermostable bacterial lipases identified by functional metagenomic screenings. Metagenomic libraries were constructed from enrichment cultures maintained at 65 to 75°C and screened resulting in the identification of initially 10 clones with lipolytic activities. Subsequently, two ORFs were identified encoding lipases, LipS and LipT. Comparative sequence analyses suggested that both enzymes are members of novel lipase families. LipS is a 30.2 kDa protein and revealed a half-life of 48 h at 70°C. The lipT gene encoded for a multimeric enzyme with a half-life of 3 h at 70°C. LipS had an optimum temperature at 70°C and LipT at 75°C. Both enzymes catalyzed hydrolysis of long-chain (C12 and C14) fatty acid esters and additionally hydrolyzed a number of industry-relevant substrates. LipS was highly specific for (R)-ibuprofen-phenyl ester with an enantiomeric excess (ee) of 99%. Furthermore, LipS was able to synthesize 1-propyl laurate and 1-tetradecyl myristate at 70°C with rates similar to those of the lipase CalB from Candida antarctica. LipS represents the first example of a thermostable metagenome-derived lipase with significant synthesis activities. Its X-ray structure was solved with a resolution of 1.99 Å revealing an unusually compact lid structure. |
format |
Article in Journal/Newspaper |
author |
Chow, Jennifer Kovacic, Filip Menyes, Ina Bornscheuer, Uwe T. Eckstein, Marrit Thum, Oliver Liese, Andreas Mueller-Dieckmann, Jochen Jaeger, Karl-Erich Streit, Wolfgang R. Dall Antonia, Yuliya Krauss, Ulrich Fersini, Francesco Schmeisser, Christel Lauinger, Benjamin Bongen, Patrick Pietruszka, Jörg Schmidt, Marlen |
author_facet |
Chow, Jennifer Kovacic, Filip Menyes, Ina Bornscheuer, Uwe T. Eckstein, Marrit Thum, Oliver Liese, Andreas Mueller-Dieckmann, Jochen Jaeger, Karl-Erich Streit, Wolfgang R. Dall Antonia, Yuliya Krauss, Ulrich Fersini, Francesco Schmeisser, Christel Lauinger, Benjamin Bongen, Patrick Pietruszka, Jörg Schmidt, Marlen |
author_sort |
Chow, Jennifer |
title |
The Metagenome-Derived Enzymes LipS and LipT Increase the Diversity of Known Lipases |
title_short |
The Metagenome-Derived Enzymes LipS and LipT Increase the Diversity of Known Lipases |
title_full |
The Metagenome-Derived Enzymes LipS and LipT Increase the Diversity of Known Lipases |
title_fullStr |
The Metagenome-Derived Enzymes LipS and LipT Increase the Diversity of Known Lipases |
title_full_unstemmed |
The Metagenome-Derived Enzymes LipS and LipT Increase the Diversity of Known Lipases |
title_sort |
metagenome-derived enzymes lips and lipt increase the diversity of known lipases |
publisher |
PLoS |
publishDate |
2012 |
url |
https://juser.fz-juelich.de/record/202186 https://juser.fz-juelich.de/search?p=id:%22FZJ-2015-04475%22 |
op_coverage |
DE |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
PLoS one 7(10), e47665 - (2012). doi:10.1371/journal.pone.0047665 |
op_relation |
info:eu-repo/semantics/altIdentifier/wos/WOS:000310310200064 info:eu-repo/semantics/altIdentifier/issn/1932-6203 info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0047665 info:eu-repo/semantics/altIdentifier/hdl/2128/8945 https://juser.fz-juelich.de/record/202186 https://juser.fz-juelich.de/search?p=id:%22FZJ-2015-04475%22 |
op_rights |
info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1371/journal.pone.0047665 |
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PLoS ONE |
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7 |
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10 |
container_start_page |
e47665 |
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1766150617310429184 |