Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9

Detalhes bibliográficos
Autor(a) principal: Greiner,Ralf
Data de Publicação: 2009
Outros Autores: Silva,Lucineia Gomes da, Couri,Sonia
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Journal of Microbiology
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822009000400010
Resumo: An extracellular phytase from Aspergillus niger 11T53A9 was purified about 51-fold to apparent homogeneity with a recovery of 20.3% referred to the phytase activity in the crude extract. Purification was achieved by ammonium sulphate precipitation, ion chromataography and gel filtration. The purified enzyme behaved as a monomeric protein with a molecular mass of about 85 kDa and exhibited maximal phytate-degrading activity at pH 5.0. Optimum temperature for the degradation of phytate was 55°C. The kinetic parameters for the hydrolysis of sodium phytate were determined to be K M = 54 µmol l-1 and k cat = 190 sec-1 at pH 5.0 and 37°C. The purified enzyme was rather specific for phytate dephosphorylation. It was shown that the phytase preferably dephosphorylates myo-inositol hexakisphosphate in a stereospecific way by sequential removal of phosphate groups via D-Ins(1,2,4,5,6)P5, D-Ins(1,2,5,6)P4, D-Ins(1,2,6)P3, D-Ins(1,2)P2 to finally Ins(2)P.
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spelling Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9Aspergillus nigerphytate-degrading enzymephytatephytaseAn extracellular phytase from Aspergillus niger 11T53A9 was purified about 51-fold to apparent homogeneity with a recovery of 20.3% referred to the phytase activity in the crude extract. Purification was achieved by ammonium sulphate precipitation, ion chromataography and gel filtration. The purified enzyme behaved as a monomeric protein with a molecular mass of about 85 kDa and exhibited maximal phytate-degrading activity at pH 5.0. Optimum temperature for the degradation of phytate was 55°C. The kinetic parameters for the hydrolysis of sodium phytate were determined to be K M = 54 µmol l-1 and k cat = 190 sec-1 at pH 5.0 and 37°C. The purified enzyme was rather specific for phytate dephosphorylation. It was shown that the phytase preferably dephosphorylates myo-inositol hexakisphosphate in a stereospecific way by sequential removal of phosphate groups via D-Ins(1,2,4,5,6)P5, D-Ins(1,2,5,6)P4, D-Ins(1,2,6)P3, D-Ins(1,2)P2 to finally Ins(2)P.Sociedade Brasileira de Microbiologia2009-12-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822009000400010Brazilian Journal of Microbiology v.40 n.4 2009reponame:Brazilian Journal of Microbiologyinstname:Sociedade Brasileira de Microbiologia (SBM)instacron:SBM10.1590/S1517-83822009000400010info:eu-repo/semantics/openAccessGreiner,RalfSilva,Lucineia Gomes daCouri,Soniaeng2009-10-06T00:00:00Zoai:scielo:S1517-83822009000400010Revistahttps://www.scielo.br/j/bjm/ONGhttps://old.scielo.br/oai/scielo-oai.phpbjm@sbmicrobiologia.org.br||mbmartin@usp.br1678-44051517-8382opendoar:2009-10-06T00:00Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)false
dc.title.none.fl_str_mv Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9
title Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9
spellingShingle Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9
Greiner,Ralf
Aspergillus niger
phytate-degrading enzyme
phytate
phytase
title_short Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9
title_full Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9
title_fullStr Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9
title_full_unstemmed Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9
title_sort Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9
author Greiner,Ralf
author_facet Greiner,Ralf
Silva,Lucineia Gomes da
Couri,Sonia
author_role author
author2 Silva,Lucineia Gomes da
Couri,Sonia
author2_role author
author
dc.contributor.author.fl_str_mv Greiner,Ralf
Silva,Lucineia Gomes da
Couri,Sonia
dc.subject.por.fl_str_mv Aspergillus niger
phytate-degrading enzyme
phytate
phytase
topic Aspergillus niger
phytate-degrading enzyme
phytate
phytase
description An extracellular phytase from Aspergillus niger 11T53A9 was purified about 51-fold to apparent homogeneity with a recovery of 20.3% referred to the phytase activity in the crude extract. Purification was achieved by ammonium sulphate precipitation, ion chromataography and gel filtration. The purified enzyme behaved as a monomeric protein with a molecular mass of about 85 kDa and exhibited maximal phytate-degrading activity at pH 5.0. Optimum temperature for the degradation of phytate was 55°C. The kinetic parameters for the hydrolysis of sodium phytate were determined to be K M = 54 µmol l-1 and k cat = 190 sec-1 at pH 5.0 and 37°C. The purified enzyme was rather specific for phytate dephosphorylation. It was shown that the phytase preferably dephosphorylates myo-inositol hexakisphosphate in a stereospecific way by sequential removal of phosphate groups via D-Ins(1,2,4,5,6)P5, D-Ins(1,2,5,6)P4, D-Ins(1,2,6)P3, D-Ins(1,2)P2 to finally Ins(2)P.
publishDate 2009
dc.date.none.fl_str_mv 2009-12-01
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=S1517-83822009000400010
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822009000400010
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S1517-83822009000400010
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 Sociedade Brasileira de Microbiologia
publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
dc.source.none.fl_str_mv Brazilian Journal of Microbiology v.40 n.4 2009
reponame:Brazilian Journal of Microbiology
instname:Sociedade Brasileira de Microbiologia (SBM)
instacron:SBM
instname_str Sociedade Brasileira de Microbiologia (SBM)
instacron_str SBM
institution SBM
reponame_str Brazilian Journal of Microbiology
collection Brazilian Journal of Microbiology
repository.name.fl_str_mv Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)
repository.mail.fl_str_mv bjm@sbmicrobiologia.org.br||mbmartin@usp.br
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