Extraction of lipase from Burkholderia cepacia by PEG/Phosphate ATPS and its biochemical characterization
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=S1516-89132012000100002 |
Resumo: | This work aimed to study the partitioning of a lipase produced by Burkholderia cepacia in PEG/Phosphate aqueous two phase system (ATPS) and its characterization. Lipase was produced by B. cepacia strains in a fermenter. Enzyme partitioning occurred at pH 6.0 and 8.0, using PEG 1500 and 6000 on two tie lines. Metal ions, pH and temperature effects on enzyme activity were evaluated. Five milliliter of 7.5% olive oil emulsion with 2.5% gumarabic in 0.1M sodium phosphate buffer at pH 8.0 and 37ºC were used for the activity determinations. Results showed that crude stratum from B. cepacia was partitioned by PEG1500/phosphate ATPS at pH 6.0 or 8.0 for, which the partitioning coefficients were 108-and 209-folds. Lipase presented optimal activity conditions at 37ºC and pH 8.0; it showed pH-stability for 4 h of incubation at different pH values at 37ºC. Metal ions such as Mn2+ , Co2+, I-and Ca2+ sustained enzymatic activities; however, it was inhibited by the presence of Fe2+, Hg2+ and Al3+ . Km and Vmax values were 0.258 U/mg and 43.90 g/L, respectively. A molecular weight of 33 kDa and an isoelectric point at pH 5.0 were determined by SDS-PAGE and IFS electrophoresis, respectively. |
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Brazilian Archives of Biology and Technology |
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Extraction of lipase from Burkholderia cepacia by PEG/Phosphate ATPS and its biochemical characterizationlipaseBurkholderia cepaciaaqueous two phase systempartitioningcharacterizationThis work aimed to study the partitioning of a lipase produced by Burkholderia cepacia in PEG/Phosphate aqueous two phase system (ATPS) and its characterization. Lipase was produced by B. cepacia strains in a fermenter. Enzyme partitioning occurred at pH 6.0 and 8.0, using PEG 1500 and 6000 on two tie lines. Metal ions, pH and temperature effects on enzyme activity were evaluated. Five milliliter of 7.5% olive oil emulsion with 2.5% gumarabic in 0.1M sodium phosphate buffer at pH 8.0 and 37ºC were used for the activity determinations. Results showed that crude stratum from B. cepacia was partitioned by PEG1500/phosphate ATPS at pH 6.0 or 8.0 for, which the partitioning coefficients were 108-and 209-folds. Lipase presented optimal activity conditions at 37ºC and pH 8.0; it showed pH-stability for 4 h of incubation at different pH values at 37ºC. Metal ions such as Mn2+ , Co2+, I-and Ca2+ sustained enzymatic activities; however, it was inhibited by the presence of Fe2+, Hg2+ and Al3+ . Km and Vmax values were 0.258 U/mg and 43.90 g/L, respectively. A molecular weight of 33 kDa and an isoelectric point at pH 5.0 were determined by SDS-PAGE and IFS electrophoresis, respectively.Instituto de Tecnologia do Paraná - Tecpar2012-02-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132012000100002Brazilian Archives of Biology and Technology v.55 n.1 2012reponame:Brazilian Archives of Biology and Technologyinstname:Instituto de Tecnologia do Paraná (Tecpar)instacron:TECPAR10.1590/S1516-89132012000100002info:eu-repo/semantics/openAccessPadilha,Giovana da SilvaSantana,José Carlos CurveloAlegre,Ranulfo MonteTambourgi,Elias Basileeng2012-03-29T00:00:00Zoai:scielo:S1516-89132012000100002Revistahttps://www.scielo.br/j/babt/https://old.scielo.br/oai/scielo-oai.phpbabt@tecpar.br||babt@tecpar.br1678-43241516-8913opendoar:2012-03-29T00:00Brazilian Archives of Biology and Technology - Instituto de Tecnologia do Paraná (Tecpar)false |
dc.title.none.fl_str_mv |
Extraction of lipase from Burkholderia cepacia by PEG/Phosphate ATPS and its biochemical characterization |
title |
Extraction of lipase from Burkholderia cepacia by PEG/Phosphate ATPS and its biochemical characterization |
spellingShingle |
Extraction of lipase from Burkholderia cepacia by PEG/Phosphate ATPS and its biochemical characterization Padilha,Giovana da Silva lipase Burkholderia cepacia aqueous two phase system partitioning characterization |
title_short |
Extraction of lipase from Burkholderia cepacia by PEG/Phosphate ATPS and its biochemical characterization |
title_full |
Extraction of lipase from Burkholderia cepacia by PEG/Phosphate ATPS and its biochemical characterization |
title_fullStr |
Extraction of lipase from Burkholderia cepacia by PEG/Phosphate ATPS and its biochemical characterization |
title_full_unstemmed |
Extraction of lipase from Burkholderia cepacia by PEG/Phosphate ATPS and its biochemical characterization |
title_sort |
Extraction of lipase from Burkholderia cepacia by PEG/Phosphate ATPS and its biochemical characterization |
author |
Padilha,Giovana da Silva |
author_facet |
Padilha,Giovana da Silva Santana,José Carlos Curvelo Alegre,Ranulfo Monte Tambourgi,Elias Basile |
author_role |
author |
author2 |
Santana,José Carlos Curvelo Alegre,Ranulfo Monte Tambourgi,Elias Basile |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Padilha,Giovana da Silva Santana,José Carlos Curvelo Alegre,Ranulfo Monte Tambourgi,Elias Basile |
dc.subject.por.fl_str_mv |
lipase Burkholderia cepacia aqueous two phase system partitioning characterization |
topic |
lipase Burkholderia cepacia aqueous two phase system partitioning characterization |
description |
This work aimed to study the partitioning of a lipase produced by Burkholderia cepacia in PEG/Phosphate aqueous two phase system (ATPS) and its characterization. Lipase was produced by B. cepacia strains in a fermenter. Enzyme partitioning occurred at pH 6.0 and 8.0, using PEG 1500 and 6000 on two tie lines. Metal ions, pH and temperature effects on enzyme activity were evaluated. Five milliliter of 7.5% olive oil emulsion with 2.5% gumarabic in 0.1M sodium phosphate buffer at pH 8.0 and 37ºC were used for the activity determinations. Results showed that crude stratum from B. cepacia was partitioned by PEG1500/phosphate ATPS at pH 6.0 or 8.0 for, which the partitioning coefficients were 108-and 209-folds. Lipase presented optimal activity conditions at 37ºC and pH 8.0; it showed pH-stability for 4 h of incubation at different pH values at 37ºC. Metal ions such as Mn2+ , Co2+, I-and Ca2+ sustained enzymatic activities; however, it was inhibited by the presence of Fe2+, Hg2+ and Al3+ . Km and Vmax values were 0.258 U/mg and 43.90 g/L, respectively. A molecular weight of 33 kDa and an isoelectric point at pH 5.0 were determined by SDS-PAGE and IFS electrophoresis, respectively. |
publishDate |
2012 |
dc.date.none.fl_str_mv |
2012-02-01 |
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=S1516-89132012000100002 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132012000100002 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S1516-89132012000100002 |
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.1 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_ |
1750318274886762496 |