The hormone-sensitive lipase from Psychrobacter sp. TA144: New insight in the structural/functional characterization
Cold-adapted esterases and lipases have been found to be dominant activities throughout the cold marine environment, indicating their importance in bacterial degradation of the organic matter. lip2 Gene from Psychrobacter sp. TA144, a micro-organism isolated from the Antarctic sea water, was cloned...
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Online Access: | http://hdl.handle.net/11588/366788 https://doi.org/10.1016/j.biochi.2010.04.001 http://www.sciencedirect.com/science/article/B6VRJ-4YTFBVF-1/2/1bc4b640d036f4d7af0e0e03f69b90b3 |
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ftunivnapoliiris:oai:www.iris.unina.it:11588/366788 2024-09-09T19:10:04+00:00 The hormone-sensitive lipase from Psychrobacter sp. TA144: New insight in the structural/functional characterization C. DE SANTI L. MANDRICH M. GIULIANI D. DE PASCALE TUTINO, MARIA LUISA PARRILLI, ERMENEGILDA DEL VECCHIO, POMPEA GIUSEPPINA GRAZIA C., DE SANTI Tutino, MARIA LUISA L., Mandrich M., Giuliani Parrilli, Ermenegilda DEL VECCHIO, POMPEA GIUSEPPINA GRAZIA D., DE PASCALE 2010 STAMPA http://hdl.handle.net/11588/366788 https://doi.org/10.1016/j.biochi.2010.04.001 http://www.sciencedirect.com/science/article/B6VRJ-4YTFBVF-1/2/1bc4b640d036f4d7af0e0e03f69b90b3 eng eng info:eu-repo/semantics/altIdentifier/pmid/20382198 info:eu-repo/semantics/altIdentifier/wos/WOS:000280570800005 volume:92 firstpage:949 lastpage:957 numberofpages:9 journal:BIOCHIMIE http://hdl.handle.net/11588/366788 doi:10.1016/j.biochi.2010.04.001 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-77954658506 http://www.sciencedirect.com/science/article/B6VRJ-4YTFBVF-1/2/1bc4b640d036f4d7af0e0e03f69b90b3 Psychrophilic micro-organism Cold-active enzyme [alpha]/[beta] hydrolase fold Ester synthesi Esterase Lipases info:eu-repo/semantics/article 2010 ftunivnapoliiris https://doi.org/10.1016/j.biochi.2010.04.001 2024-06-17T15:19:26Z Cold-adapted esterases and lipases have been found to be dominant activities throughout the cold marine environment, indicating their importance in bacterial degradation of the organic matter. lip2 Gene from Psychrobacter sp. TA144, a micro-organism isolated from the Antarctic sea water, was cloned and over-expressed in Escherichia coli. The recombinant protein (PsyHSL) accumulated in the insoluble fraction from which it was recovered in active form, purified to homogeneity and deeply characterised. Temperature dependence of PsyHSL activity was typical of psychrophilic enzymes, with an optimal temperature of 35 °C at pH 8.0. The enzyme resulted to be active on pNP-esters of fatty acids with acyl chain length from C2 to C12 and the preferred substrate was pNP-pentanoate showing a kcat = 26.2 ± 0.1 s−1, KM = 0.122 ± 0.006 mM and a kcat/KM = 215 ± 11 mM−1 s−1. The enzyme was strongly inhibited by Hg2+, Zn2+, Cu2+, Fe3+, Mn2+ ions and it resulted to be activated in presence of methanol and acetonitrile, with calculated C50 values of 1.98 M and 0.92 M, respectively. The region surrounding PsyHSL catalytic site showed an unexpected homology with the human HSL. Further, both enzymes are characterised by the presence of an extra N-terminal domain, which role in the human protein has been related to regulative function. To clarify the function of PsyHSL N-terminal domain, a 97 amino acids deleted version of the enzyme was produced in E. coli in soluble form, purified and characterised. This mutant was inactive towards all tested substrates, indicating the involvement of this region in the catalytic process. Article in Journal/Newspaper Antarc* Antarctic IRIS Università degli Studi di Napoli Federico II Antarctic The Antarctic Biochimie 92 8 949 957 |
institution |
Open Polar |
collection |
IRIS Università degli Studi di Napoli Federico II |
op_collection_id |
ftunivnapoliiris |
language |
English |
topic |
Psychrophilic micro-organism Cold-active enzyme [alpha]/[beta] hydrolase fold Ester synthesi Esterase Lipases |
spellingShingle |
Psychrophilic micro-organism Cold-active enzyme [alpha]/[beta] hydrolase fold Ester synthesi Esterase Lipases C. DE SANTI L. MANDRICH M. GIULIANI D. DE PASCALE TUTINO, MARIA LUISA PARRILLI, ERMENEGILDA DEL VECCHIO, POMPEA GIUSEPPINA GRAZIA The hormone-sensitive lipase from Psychrobacter sp. TA144: New insight in the structural/functional characterization |
topic_facet |
Psychrophilic micro-organism Cold-active enzyme [alpha]/[beta] hydrolase fold Ester synthesi Esterase Lipases |
description |
Cold-adapted esterases and lipases have been found to be dominant activities throughout the cold marine environment, indicating their importance in bacterial degradation of the organic matter. lip2 Gene from Psychrobacter sp. TA144, a micro-organism isolated from the Antarctic sea water, was cloned and over-expressed in Escherichia coli. The recombinant protein (PsyHSL) accumulated in the insoluble fraction from which it was recovered in active form, purified to homogeneity and deeply characterised. Temperature dependence of PsyHSL activity was typical of psychrophilic enzymes, with an optimal temperature of 35 °C at pH 8.0. The enzyme resulted to be active on pNP-esters of fatty acids with acyl chain length from C2 to C12 and the preferred substrate was pNP-pentanoate showing a kcat = 26.2 ± 0.1 s−1, KM = 0.122 ± 0.006 mM and a kcat/KM = 215 ± 11 mM−1 s−1. The enzyme was strongly inhibited by Hg2+, Zn2+, Cu2+, Fe3+, Mn2+ ions and it resulted to be activated in presence of methanol and acetonitrile, with calculated C50 values of 1.98 M and 0.92 M, respectively. The region surrounding PsyHSL catalytic site showed an unexpected homology with the human HSL. Further, both enzymes are characterised by the presence of an extra N-terminal domain, which role in the human protein has been related to regulative function. To clarify the function of PsyHSL N-terminal domain, a 97 amino acids deleted version of the enzyme was produced in E. coli in soluble form, purified and characterised. This mutant was inactive towards all tested substrates, indicating the involvement of this region in the catalytic process. |
author2 |
C., DE SANTI Tutino, MARIA LUISA L., Mandrich M., Giuliani Parrilli, Ermenegilda DEL VECCHIO, POMPEA GIUSEPPINA GRAZIA D., DE PASCALE |
format |
Article in Journal/Newspaper |
author |
C. DE SANTI L. MANDRICH M. GIULIANI D. DE PASCALE TUTINO, MARIA LUISA PARRILLI, ERMENEGILDA DEL VECCHIO, POMPEA GIUSEPPINA GRAZIA |
author_facet |
C. DE SANTI L. MANDRICH M. GIULIANI D. DE PASCALE TUTINO, MARIA LUISA PARRILLI, ERMENEGILDA DEL VECCHIO, POMPEA GIUSEPPINA GRAZIA |
author_sort |
C. DE SANTI |
title |
The hormone-sensitive lipase from Psychrobacter sp. TA144: New insight in the structural/functional characterization |
title_short |
The hormone-sensitive lipase from Psychrobacter sp. TA144: New insight in the structural/functional characterization |
title_full |
The hormone-sensitive lipase from Psychrobacter sp. TA144: New insight in the structural/functional characterization |
title_fullStr |
The hormone-sensitive lipase from Psychrobacter sp. TA144: New insight in the structural/functional characterization |
title_full_unstemmed |
The hormone-sensitive lipase from Psychrobacter sp. TA144: New insight in the structural/functional characterization |
title_sort |
hormone-sensitive lipase from psychrobacter sp. ta144: new insight in the structural/functional characterization |
publishDate |
2010 |
url |
http://hdl.handle.net/11588/366788 https://doi.org/10.1016/j.biochi.2010.04.001 http://www.sciencedirect.com/science/article/B6VRJ-4YTFBVF-1/2/1bc4b640d036f4d7af0e0e03f69b90b3 |
geographic |
Antarctic The Antarctic |
geographic_facet |
Antarctic The Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_relation |
info:eu-repo/semantics/altIdentifier/pmid/20382198 info:eu-repo/semantics/altIdentifier/wos/WOS:000280570800005 volume:92 firstpage:949 lastpage:957 numberofpages:9 journal:BIOCHIMIE http://hdl.handle.net/11588/366788 doi:10.1016/j.biochi.2010.04.001 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-77954658506 http://www.sciencedirect.com/science/article/B6VRJ-4YTFBVF-1/2/1bc4b640d036f4d7af0e0e03f69b90b3 |
op_doi |
https://doi.org/10.1016/j.biochi.2010.04.001 |
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Biochimie |
container_volume |
92 |
container_issue |
8 |
container_start_page |
949 |
op_container_end_page |
957 |
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