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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ftdatacite:10.15480/882.2297 2023-05-15T13:33:12+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, Jörg Schmidt, Marlen Menyes, Ina Bornscheuer, Uwe Theo Eckstein, Marrit Friederike Thum, Oliver Liese, Andreas Müller-Dieckmann, Jochen Jaeger, Karl-Erich Streit, Wolfgang R. 2012 https://dx.doi.org/10.15480/882.2297 https://tore.tuhh.de/handle/11420/2830 en eng PLOS https://dx.doi.org/10.1371/journal.pone.0047665 Creative Commons Attribution 4.0 International https://creativecommons.org/licenses/by/4.0/legalcode cc-by-4.0 info:eu-repo/semantics/openAccess CC-BY 540 Chemie 570 Biowissenschaften, Biologie 620 Ingenieurwissenschaften 570 540 620 Journal Article article-journal Text ScholarlyArticle 2012 ftdatacite https://doi.org/10.15480/882.2297 https://doi.org/10.1371/journal.pone.0047665 2022-02-08T12:55:18Z 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. © 2012 Chow et al. Text Antarc* Antarctica DataCite Metadata Store (German National Library of Science and Technology) |
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Open Polar |
collection |
DataCite Metadata Store (German National Library of Science and Technology) |
op_collection_id |
ftdatacite |
language |
English |
topic |
540 Chemie 570 Biowissenschaften, Biologie 620 Ingenieurwissenschaften 570 540 620 |
spellingShingle |
540 Chemie 570 Biowissenschaften, Biologie 620 Ingenieurwissenschaften 570 540 620 Chow, Jennifer Kovacic, Filip Dall Antonia, Yuliya Krauss, Ulrich Fersini, Francesco Schmeißer, Christel Lauinger, Benjamin Bongen, Patrick Pietruszka, Jörg Schmidt, Marlen Menyes, Ina Bornscheuer, Uwe Theo Eckstein, Marrit Friederike Thum, Oliver Liese, Andreas Müller-Dieckmann, Jochen Jaeger, Karl-Erich Streit, Wolfgang R. The Metagenome-derived enzymes LipS and LipT increase the diversity of known lipases |
topic_facet |
540 Chemie 570 Biowissenschaften, Biologie 620 Ingenieurwissenschaften 570 540 620 |
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. © 2012 Chow et al. |
format |
Text |
author |
Chow, Jennifer Kovacic, Filip Dall Antonia, Yuliya Krauss, Ulrich Fersini, Francesco Schmeißer, Christel Lauinger, Benjamin Bongen, Patrick Pietruszka, Jörg Schmidt, Marlen Menyes, Ina Bornscheuer, Uwe Theo Eckstein, Marrit Friederike Thum, Oliver Liese, Andreas Müller-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, Jörg Schmidt, Marlen Menyes, Ina Bornscheuer, Uwe Theo Eckstein, Marrit Friederike Thum, Oliver Liese, Andreas Müller-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 |
PLOS |
publishDate |
2012 |
url |
https://dx.doi.org/10.15480/882.2297 https://tore.tuhh.de/handle/11420/2830 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
https://dx.doi.org/10.1371/journal.pone.0047665 |
op_rights |
Creative Commons Attribution 4.0 International https://creativecommons.org/licenses/by/4.0/legalcode cc-by-4.0 info:eu-repo/semantics/openAccess |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.15480/882.2297 https://doi.org/10.1371/journal.pone.0047665 |
_version_ |
1766039503904964608 |