Aspects on the catalysis of lipase from porcine pancreas (type VI-s) in aqueous media: development of ion-pairs
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Brazilian Archives of Biology and Technology |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132012000200007 |
Resumo: | This article reports a first contribution for the elucidation of catalytic mechanism of Lipase from porcine pancreas, type VI-s (PPL), in hydrolyzing an ester substrate in aqueous media. The conclusions were based on the pH-profiles of Michaelis-Menten parameters k cat/Km, k cat and Km, as well as on the absolute temperature profile of k cat/Km, obtained during the hydrolysis of p-nitrophenyl laurate by PPL. It was found that (a) PPL performs catalysis by means of ion pairs formed either as Ser152-Ο-/His263-Im+H and/or Carbonyl-Ο-/His263-Im+H, (b) the parameter k cat/Km equals to k1 and thus ES is formed and destroyed in the course of a series of consecutive reactions governed by the dynamic constant K S = k2/k1, and (c) the hydrolysis of substrate is assisted by a hydrogen bond developed between deprotonated Asp176 and the positively charged imidazole of His263 across a pKa-value 3.85, necessary for efficient catalysis. |
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Aspects on the catalysis of lipase from porcine pancreas (type VI-s) in aqueous media: development of ion-pairsPorcine Pancreas Lipasemechanism of hydrolysisp-nitrophenyl laurateThis article reports a first contribution for the elucidation of catalytic mechanism of Lipase from porcine pancreas, type VI-s (PPL), in hydrolyzing an ester substrate in aqueous media. The conclusions were based on the pH-profiles of Michaelis-Menten parameters k cat/Km, k cat and Km, as well as on the absolute temperature profile of k cat/Km, obtained during the hydrolysis of p-nitrophenyl laurate by PPL. It was found that (a) PPL performs catalysis by means of ion pairs formed either as Ser152-Ο-/His263-Im+H and/or Carbonyl-Ο-/His263-Im+H, (b) the parameter k cat/Km equals to k1 and thus ES is formed and destroyed in the course of a series of consecutive reactions governed by the dynamic constant K S = k2/k1, and (c) the hydrolysis of substrate is assisted by a hydrogen bond developed between deprotonated Asp176 and the positively charged imidazole of His263 across a pKa-value 3.85, necessary for efficient catalysis.Instituto de Tecnologia do Paraná - Tecpar2012-04-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132012000200007Brazilian Archives of Biology and Technology v.55 n.2 2012reponame:Brazilian Archives of Biology and Technologyinstname:Instituto de Tecnologia do Paraná (Tecpar)instacron:TECPAR10.1590/S1516-89132012000200007info:eu-repo/semantics/openAccessKokkinou,MarinaTheodorou,Leonidas G.Papamichael,Emmanuel M.eng2012-05-03T00:00:00Zoai:scielo:S1516-89132012000200007Revistahttps://www.scielo.br/j/babt/https://old.scielo.br/oai/scielo-oai.phpbabt@tecpar.br||babt@tecpar.br1678-43241516-8913opendoar:2012-05-03T00:00Brazilian Archives of Biology and Technology - Instituto de Tecnologia do Paraná (Tecpar)false |
dc.title.none.fl_str_mv |
Aspects on the catalysis of lipase from porcine pancreas (type VI-s) in aqueous media: development of ion-pairs |
title |
Aspects on the catalysis of lipase from porcine pancreas (type VI-s) in aqueous media: development of ion-pairs |
spellingShingle |
Aspects on the catalysis of lipase from porcine pancreas (type VI-s) in aqueous media: development of ion-pairs Kokkinou,Marina Porcine Pancreas Lipase mechanism of hydrolysis p-nitrophenyl laurate |
title_short |
Aspects on the catalysis of lipase from porcine pancreas (type VI-s) in aqueous media: development of ion-pairs |
title_full |
Aspects on the catalysis of lipase from porcine pancreas (type VI-s) in aqueous media: development of ion-pairs |
title_fullStr |
Aspects on the catalysis of lipase from porcine pancreas (type VI-s) in aqueous media: development of ion-pairs |
title_full_unstemmed |
Aspects on the catalysis of lipase from porcine pancreas (type VI-s) in aqueous media: development of ion-pairs |
title_sort |
Aspects on the catalysis of lipase from porcine pancreas (type VI-s) in aqueous media: development of ion-pairs |
author |
Kokkinou,Marina |
author_facet |
Kokkinou,Marina Theodorou,Leonidas G. Papamichael,Emmanuel M. |
author_role |
author |
author2 |
Theodorou,Leonidas G. Papamichael,Emmanuel M. |
author2_role |
author author |
dc.contributor.author.fl_str_mv |
Kokkinou,Marina Theodorou,Leonidas G. Papamichael,Emmanuel M. |
dc.subject.por.fl_str_mv |
Porcine Pancreas Lipase mechanism of hydrolysis p-nitrophenyl laurate |
topic |
Porcine Pancreas Lipase mechanism of hydrolysis p-nitrophenyl laurate |
description |
This article reports a first contribution for the elucidation of catalytic mechanism of Lipase from porcine pancreas, type VI-s (PPL), in hydrolyzing an ester substrate in aqueous media. The conclusions were based on the pH-profiles of Michaelis-Menten parameters k cat/Km, k cat and Km, as well as on the absolute temperature profile of k cat/Km, obtained during the hydrolysis of p-nitrophenyl laurate by PPL. It was found that (a) PPL performs catalysis by means of ion pairs formed either as Ser152-Ο-/His263-Im+H and/or Carbonyl-Ο-/His263-Im+H, (b) the parameter k cat/Km equals to k1 and thus ES is formed and destroyed in the course of a series of consecutive reactions governed by the dynamic constant K S = k2/k1, and (c) the hydrolysis of substrate is assisted by a hydrogen bond developed between deprotonated Asp176 and the positively charged imidazole of His263 across a pKa-value 3.85, necessary for efficient catalysis. |
publishDate |
2012 |
dc.date.none.fl_str_mv |
2012-04-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132012000200007 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132012000200007 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S1516-89132012000200007 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Instituto de Tecnologia do Paraná - Tecpar |
publisher.none.fl_str_mv |
Instituto de Tecnologia do Paraná - Tecpar |
dc.source.none.fl_str_mv |
Brazilian Archives of Biology and Technology v.55 n.2 2012 reponame:Brazilian Archives of Biology and Technology instname:Instituto de Tecnologia do Paraná (Tecpar) instacron:TECPAR |
instname_str |
Instituto de Tecnologia do Paraná (Tecpar) |
instacron_str |
TECPAR |
institution |
TECPAR |
reponame_str |
Brazilian Archives of Biology and Technology |
collection |
Brazilian Archives of Biology and Technology |
repository.name.fl_str_mv |
Brazilian Archives of Biology and Technology - Instituto de Tecnologia do Paraná (Tecpar) |
repository.mail.fl_str_mv |
babt@tecpar.br||babt@tecpar.br |
_version_ |
1750318275179315200 |