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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2012
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oai:doaj.org-article:1ddb1c408a7041adbcaf448fed9090bf2021-11-18T08:11:12ZThe metagenome-derived enzymes LipS and LipT increase the diversity of known lipases.1932-620310.1371/journal.pone.0047665https://doaj.org/article/1ddb1c408a7041adbcaf448fed9090bf2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23112831/?tool=EBIhttps://doaj.org/toc/1932-6203Triacylglycerol 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 (C(12) and C(14)) 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.Jennifer ChowFilip KovacicYuliya Dall AntoniaUlrich KraussFrancesco FersiniChristel SchmeisserBenjamin LauingerPatrick BongenJoerg PietruszkaMarlen SchmidtIna MenyesUwe T BornscheuerMarrit EcksteinOliver ThumAndreas LieseJochen Mueller-DieckmannKarl-Erich JaegerWolfgang R StreitPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 10, p e47665 (2012) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
Medicine R Science Q |
| spellingShingle |
Medicine R Science Q Jennifer Chow Filip Kovacic Yuliya Dall Antonia Ulrich Krauss Francesco Fersini Christel Schmeisser Benjamin Lauinger Patrick Bongen Joerg Pietruszka Marlen Schmidt Ina Menyes Uwe T Bornscheuer Marrit Eckstein Oliver Thum Andreas Liese Jochen Mueller-Dieckmann Karl-Erich Jaeger Wolfgang R Streit The metagenome-derived enzymes LipS and LipT increase the diversity of known lipases. |
| 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 (C(12) and C(14)) 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 |
| author |
Jennifer Chow Filip Kovacic Yuliya Dall Antonia Ulrich Krauss Francesco Fersini Christel Schmeisser Benjamin Lauinger Patrick Bongen Joerg Pietruszka Marlen Schmidt Ina Menyes Uwe T Bornscheuer Marrit Eckstein Oliver Thum Andreas Liese Jochen Mueller-Dieckmann Karl-Erich Jaeger Wolfgang R Streit |
| author_facet |
Jennifer Chow Filip Kovacic Yuliya Dall Antonia Ulrich Krauss Francesco Fersini Christel Schmeisser Benjamin Lauinger Patrick Bongen Joerg Pietruszka Marlen Schmidt Ina Menyes Uwe T Bornscheuer Marrit Eckstein Oliver Thum Andreas Liese Jochen Mueller-Dieckmann Karl-Erich Jaeger Wolfgang R Streit |
| author_sort |
Jennifer Chow |
| 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 (PLoS) |
| publishDate |
2012 |
| url |
https://doaj.org/article/1ddb1c408a7041adbcaf448fed9090bf |
| work_keys_str_mv |
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