Aspects on the catalysis of lipase from porcine pancreas (type VI-s) in aqueous media: development of ion-pairs

Detalhes bibliográficos
Autor(a) principal: Kokkinou,Marina
Data de Publicação: 2012
Outros Autores: Theodorou,Leonidas G., Papamichael,Emmanuel M.
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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spelling 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
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