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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ftpubmed:oai:pubmedcentral.nih.gov:3480424 2023-05-15T14:04:04+02:00 The Metagenome-Derived Enzymes LipS and LipT Increase the Diversity of Known Lipases Chow, Jennifer Kovacic, Filip Dall Antonia, Yuliya Krauss, Ulrich Fersini, Francesco Schmeisser, Christel Lauinger, Benjamin Bongen, Patrick Pietruszka, Joerg Schmidt, Marlen Menyes, Ina Bornscheuer, Uwe T. Eckstein, Marrit Thum, Oliver Liese, Andreas Mueller-Dieckmann, Jochen Jaeger, Karl-Erich Streit, Wolfgang R. 2012-10-24 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3480424 http://www.ncbi.nlm.nih.gov/pubmed/23112831 https://doi.org/10.1371/journal.pone.0047665 en eng Public Library of Science http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3480424 http://www.ncbi.nlm.nih.gov/pubmed/23112831 http://dx.doi.org/10.1371/journal.pone.0047665 This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. CC-BY Research Article Text 2012 ftpubmed https://doi.org/10.1371/journal.pone.0047665 2013-09-04T14:57:19Z 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. Text Antarc* Antarctica PubMed Central (PMC) PLoS ONE 7 10 e47665 |
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English |
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Research Article |
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Research Article Chow, Jennifer Kovacic, Filip Dall Antonia, Yuliya Krauss, Ulrich Fersini, Francesco Schmeisser, Christel Lauinger, Benjamin Bongen, Patrick Pietruszka, Joerg Schmidt, Marlen Menyes, Ina Bornscheuer, Uwe T. Eckstein, Marrit Thum, Oliver Liese, Andreas Mueller-Dieckmann, Jochen Jaeger, Karl-Erich Streit, Wolfgang R. The Metagenome-Derived Enzymes LipS and LipT Increase the Diversity of Known Lipases |
topic_facet |
Research Article |
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 |
Text |
author |
Chow, Jennifer Kovacic, Filip Dall Antonia, Yuliya Krauss, Ulrich Fersini, Francesco Schmeisser, Christel Lauinger, Benjamin Bongen, Patrick Pietruszka, Joerg Schmidt, Marlen Menyes, Ina Bornscheuer, Uwe T. Eckstein, Marrit Thum, Oliver Liese, Andreas Mueller-Dieckmann, Jochen Jaeger, Karl-Erich Streit, Wolfgang R. |
author_facet |
Chow, Jennifer Kovacic, Filip Dall Antonia, Yuliya Krauss, Ulrich Fersini, Francesco Schmeisser, Christel Lauinger, Benjamin Bongen, Patrick Pietruszka, Joerg Schmidt, Marlen Menyes, Ina Bornscheuer, Uwe T. Eckstein, Marrit Thum, Oliver Liese, Andreas Mueller-Dieckmann, Jochen Jaeger, Karl-Erich Streit, Wolfgang R. |
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 |
Public Library of Science |
publishDate |
2012 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3480424 http://www.ncbi.nlm.nih.gov/pubmed/23112831 https://doi.org/10.1371/journal.pone.0047665 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3480424 http://www.ncbi.nlm.nih.gov/pubmed/23112831 http://dx.doi.org/10.1371/journal.pone.0047665 |
op_rights |
This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
op_rightsnorm |
CC-BY |
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 |
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e47665 |
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