Cloning and fusion expression of a cold-active lipase from marine antarctic origin
Antarctic seawater bacteria producing extracellular lipolytic enzymes with activity at low temperatures were isolated. The most promising strain was selected to perform a 16S characterization which identified it as a Psychrobacter sp. The genomic DNA of this bacterium was used for a PCR screening us...
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ftconicyt:oai:localhost:10533/198054 2023-05-15T13:53:09+02:00 Cloning and fusion expression of a cold-active lipase from marine antarctic origin Asenjo-De Leuze, Juan Salazar-Aguirre, Oriana Acevedo-Cox, Juan Reyes-B., Fernando Andrews-Farrow, Barbara Parra-Atala, Loreto USA NEW YORK http://hdl.handle.net/10533/198054 https://doi.org/10.1016/j.enzmictec.2007.11.003 ENG eng ELSEVIER SCIENCE INC instname: Conicyt reponame: Repositorio Digital RI2.0 info:eu-repo/grantAgreement/Fondef/D04I1374 info:eu-repo/semantics/dataset/hdl.handle.net/10533/93477 http://hdl.handle.net/10533/198054 doi:10.1016/j.enzmictec.2007.11.003 wos: WOS:000254728500010 eissn: 0 issn: 0141-0229 info:eu-repo/semantics/openAccess ENZYME AND MICROBIAL TECHNOLOGY info:eu-repo/semantics/article ftconicyt https://doi.org/10.1016/j.enzmictec.2007.11.003 2019-09-07T08:18:48Z Antarctic seawater bacteria producing extracellular lipolytic enzymes with activity at low temperatures were isolated. The most promising strain was selected to perform a 16S characterization which identified it as a Psychrobacter sp. The genomic DNA of this bacterium was used for a PCR screening using primers obtained from multiple sequence alignments of lipases belonging to the hormone sensitive lipase (HSL) group of the prokaryotic species. This allowed cloning and sequencing of the DNA that partially encodes a novel lipase protein (240 bp, 80 aa). Subsequently the complete gene was obtained by a genome-walking technique. An open reading frame of 1293 bp was found, which encodes for a polypeptide of 431 amino acids, and presents 89% identity with lipase 2 from Moraxella TA 144 previously described; however its properties are very different. The promoter and downstream sequences of this gene were also obtained. The new lipase gene was cloned into expression vector pMAL-c2E and integrated into E. coli TB1. A recombinant fusion protein (MBP-lipase) with a molecular weight of 90 kDa was produced and purified which showed lipolytic activity. The optimum temperature for this fusion lipase was 20 degrees C at pH 8.0, and the activation energy was 5.5 kcal/mol between 5 and 20 degrees C at the same pH. (c) 2007 Elsevier Inc. All rights reserved. Article in Journal/Newspaper Antarc* Antarctic Repositorio Digital Conicyt RI 2.0 (Comisión Nacional de Investigación Científica y Tecnológica) Antarctic Enzyme and Microbial Technology 42 4 371 377 |
institution |
Open Polar |
collection |
Repositorio Digital Conicyt RI 2.0 (Comisión Nacional de Investigación Científica y Tecnológica) |
op_collection_id |
ftconicyt |
language |
English |
description |
Antarctic seawater bacteria producing extracellular lipolytic enzymes with activity at low temperatures were isolated. The most promising strain was selected to perform a 16S characterization which identified it as a Psychrobacter sp. The genomic DNA of this bacterium was used for a PCR screening using primers obtained from multiple sequence alignments of lipases belonging to the hormone sensitive lipase (HSL) group of the prokaryotic species. This allowed cloning and sequencing of the DNA that partially encodes a novel lipase protein (240 bp, 80 aa). Subsequently the complete gene was obtained by a genome-walking technique. An open reading frame of 1293 bp was found, which encodes for a polypeptide of 431 amino acids, and presents 89% identity with lipase 2 from Moraxella TA 144 previously described; however its properties are very different. The promoter and downstream sequences of this gene were also obtained. The new lipase gene was cloned into expression vector pMAL-c2E and integrated into E. coli TB1. A recombinant fusion protein (MBP-lipase) with a molecular weight of 90 kDa was produced and purified which showed lipolytic activity. The optimum temperature for this fusion lipase was 20 degrees C at pH 8.0, and the activation energy was 5.5 kcal/mol between 5 and 20 degrees C at the same pH. (c) 2007 Elsevier Inc. All rights reserved. |
format |
Article in Journal/Newspaper |
author |
Asenjo-De Leuze, Juan Salazar-Aguirre, Oriana Acevedo-Cox, Juan Reyes-B., Fernando Andrews-Farrow, Barbara Parra-Atala, Loreto |
spellingShingle |
Asenjo-De Leuze, Juan Salazar-Aguirre, Oriana Acevedo-Cox, Juan Reyes-B., Fernando Andrews-Farrow, Barbara Parra-Atala, Loreto Cloning and fusion expression of a cold-active lipase from marine antarctic origin |
author_facet |
Asenjo-De Leuze, Juan Salazar-Aguirre, Oriana Acevedo-Cox, Juan Reyes-B., Fernando Andrews-Farrow, Barbara Parra-Atala, Loreto |
author_sort |
Asenjo-De Leuze, Juan |
title |
Cloning and fusion expression of a cold-active lipase from marine antarctic origin |
title_short |
Cloning and fusion expression of a cold-active lipase from marine antarctic origin |
title_full |
Cloning and fusion expression of a cold-active lipase from marine antarctic origin |
title_fullStr |
Cloning and fusion expression of a cold-active lipase from marine antarctic origin |
title_full_unstemmed |
Cloning and fusion expression of a cold-active lipase from marine antarctic origin |
title_sort |
cloning and fusion expression of a cold-active lipase from marine antarctic origin |
publisher |
ELSEVIER SCIENCE INC |
url |
http://hdl.handle.net/10533/198054 https://doi.org/10.1016/j.enzmictec.2007.11.003 |
op_coverage |
USA NEW YORK |
geographic |
Antarctic |
geographic_facet |
Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
ENZYME AND MICROBIAL TECHNOLOGY |
op_relation |
instname: Conicyt reponame: Repositorio Digital RI2.0 info:eu-repo/grantAgreement/Fondef/D04I1374 info:eu-repo/semantics/dataset/hdl.handle.net/10533/93477 http://hdl.handle.net/10533/198054 doi:10.1016/j.enzmictec.2007.11.003 wos: WOS:000254728500010 eissn: 0 issn: 0141-0229 |
op_rights |
info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1016/j.enzmictec.2007.11.003 |
container_title |
Enzyme and Microbial Technology |
container_volume |
42 |
container_issue |
4 |
container_start_page |
371 |
op_container_end_page |
377 |
_version_ |
1766258132365869056 |