Extraction, partial characterization and susceptibility to Hg2+ of acid phosphatase from the microalgae Pseudokirchneriella subcapitata
Autor(a) principal: | |
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Data de Publicação: | 2009 |
Outros Autores: | |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Scientia Agrícola (Online) |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-90162009000500008 |
Resumo: | Pseudokirchneriella subcapitata is a unicellular green algae widely distributed in freshwater and soils. Due to its cosmopolitan characteristic, its use is recommended by national and international protocols in ecotoxicity studies. The alteration of phosphatase activities by agriculture pollutants like heavy metals has been extensively used as a biomarker in risk assessment and biomonitoring. In this study, we compared the extraction of acid phosphatase from P. subcapitata by different procedures and we studied the stability, substrates specificity, kinetics and the effect of Hg2+ in the crude extract. The freezing and thawing technique associated with probe sonication was the most suitable method of extraction. The enzyme was stable when frozen at -20ºC for at least six months, showed an optimum pH of 5 and a Km value of 0.27 mM for p-nitrophenylphosphate (pNPP) as substrate. Some natural organic substrates were cleaved by a similar extent as the synthetic substrate pNPP. Short term exposure (24 hours) to Hg2+ had little effect but inhibition of the specific activity was observed after 7 days with EC50 (concentration of Hg2+ that promotes 50% decrease of specific activity) value of 12.63 μM Hg2+. |
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Scientia Agrícola (Online) |
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Extraction, partial characterization and susceptibility to Hg2+ of acid phosphatase from the microalgae Pseudokirchneriella subcapitataSelenastrum capricornutumalgaeenzymemetaltoxicityPseudokirchneriella subcapitata is a unicellular green algae widely distributed in freshwater and soils. Due to its cosmopolitan characteristic, its use is recommended by national and international protocols in ecotoxicity studies. The alteration of phosphatase activities by agriculture pollutants like heavy metals has been extensively used as a biomarker in risk assessment and biomonitoring. In this study, we compared the extraction of acid phosphatase from P. subcapitata by different procedures and we studied the stability, substrates specificity, kinetics and the effect of Hg2+ in the crude extract. The freezing and thawing technique associated with probe sonication was the most suitable method of extraction. The enzyme was stable when frozen at -20ºC for at least six months, showed an optimum pH of 5 and a Km value of 0.27 mM for p-nitrophenylphosphate (pNPP) as substrate. Some natural organic substrates were cleaved by a similar extent as the synthetic substrate pNPP. Short term exposure (24 hours) to Hg2+ had little effect but inhibition of the specific activity was observed after 7 days with EC50 (concentration of Hg2+ that promotes 50% decrease of specific activity) value of 12.63 μM Hg2+.Escola Superior de Agricultura "Luiz de Queiroz"2009-10-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-90162009000500008Scientia Agricola v.66 n.5 2009reponame:Scientia Agrícola (Online)instname:Universidade de São Paulo (USP)instacron:USP10.1590/S0103-90162009000500008info:eu-repo/semantics/openAccessJonsson,Claudio MartínAoyama,Hiroshieng2009-10-05T00:00:00Zoai:scielo:S0103-90162009000500008Revistahttp://revistas.usp.br/sa/indexPUBhttps://old.scielo.br/oai/scielo-oai.phpscientia@usp.br||alleoni@usp.br1678-992X0103-9016opendoar:2009-10-05T00:00Scientia Agrícola (Online) - Universidade de São Paulo (USP)false |
dc.title.none.fl_str_mv |
Extraction, partial characterization and susceptibility to Hg2+ of acid phosphatase from the microalgae Pseudokirchneriella subcapitata |
title |
Extraction, partial characterization and susceptibility to Hg2+ of acid phosphatase from the microalgae Pseudokirchneriella subcapitata |
spellingShingle |
Extraction, partial characterization and susceptibility to Hg2+ of acid phosphatase from the microalgae Pseudokirchneriella subcapitata Jonsson,Claudio Martín Selenastrum capricornutum algae enzyme metal toxicity |
title_short |
Extraction, partial characterization and susceptibility to Hg2+ of acid phosphatase from the microalgae Pseudokirchneriella subcapitata |
title_full |
Extraction, partial characterization and susceptibility to Hg2+ of acid phosphatase from the microalgae Pseudokirchneriella subcapitata |
title_fullStr |
Extraction, partial characterization and susceptibility to Hg2+ of acid phosphatase from the microalgae Pseudokirchneriella subcapitata |
title_full_unstemmed |
Extraction, partial characterization and susceptibility to Hg2+ of acid phosphatase from the microalgae Pseudokirchneriella subcapitata |
title_sort |
Extraction, partial characterization and susceptibility to Hg2+ of acid phosphatase from the microalgae Pseudokirchneriella subcapitata |
author |
Jonsson,Claudio Martín |
author_facet |
Jonsson,Claudio Martín Aoyama,Hiroshi |
author_role |
author |
author2 |
Aoyama,Hiroshi |
author2_role |
author |
dc.contributor.author.fl_str_mv |
Jonsson,Claudio Martín Aoyama,Hiroshi |
dc.subject.por.fl_str_mv |
Selenastrum capricornutum algae enzyme metal toxicity |
topic |
Selenastrum capricornutum algae enzyme metal toxicity |
description |
Pseudokirchneriella subcapitata is a unicellular green algae widely distributed in freshwater and soils. Due to its cosmopolitan characteristic, its use is recommended by national and international protocols in ecotoxicity studies. The alteration of phosphatase activities by agriculture pollutants like heavy metals has been extensively used as a biomarker in risk assessment and biomonitoring. In this study, we compared the extraction of acid phosphatase from P. subcapitata by different procedures and we studied the stability, substrates specificity, kinetics and the effect of Hg2+ in the crude extract. The freezing and thawing technique associated with probe sonication was the most suitable method of extraction. The enzyme was stable when frozen at -20ºC for at least six months, showed an optimum pH of 5 and a Km value of 0.27 mM for p-nitrophenylphosphate (pNPP) as substrate. Some natural organic substrates were cleaved by a similar extent as the synthetic substrate pNPP. Short term exposure (24 hours) to Hg2+ had little effect but inhibition of the specific activity was observed after 7 days with EC50 (concentration of Hg2+ that promotes 50% decrease of specific activity) value of 12.63 μM Hg2+. |
publishDate |
2009 |
dc.date.none.fl_str_mv |
2009-10-01 |
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=S0103-90162009000500008 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-90162009000500008 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S0103-90162009000500008 |
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 |
Escola Superior de Agricultura "Luiz de Queiroz" |
publisher.none.fl_str_mv |
Escola Superior de Agricultura "Luiz de Queiroz" |
dc.source.none.fl_str_mv |
Scientia Agricola v.66 n.5 2009 reponame:Scientia Agrícola (Online) instname:Universidade de São Paulo (USP) instacron:USP |
instname_str |
Universidade de São Paulo (USP) |
instacron_str |
USP |
institution |
USP |
reponame_str |
Scientia Agrícola (Online) |
collection |
Scientia Agrícola (Online) |
repository.name.fl_str_mv |
Scientia Agrícola (Online) - Universidade de São Paulo (USP) |
repository.mail.fl_str_mv |
scientia@usp.br||alleoni@usp.br |
_version_ |
1748936461530431488 |