Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance
Autor(a) principal: | |
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Data de Publicação: | 2015 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://dx.doi.org/10.1590/S1517-838220120037 http://www.locus.ufv.br/handle/123456789/12735 |
Resumo: | An Aspergillus niger UFV-1 phytase was characterized and made available for industrial application. The enzyme was purified via ultrafiltration followed by acid precipitation, ion exchange and gel filtration chromatography. This protein exhibited a molecular mass of 161 kDa in gel filtration and 81 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), indicating that it may be a dimer. It presented an optimum temperature of 60 °C and optimum pH of 2.0. The K M for so- dium phytate hydrolysis was 30.9 mM, while the k cat and k cat /K M were 1.46 x10^ 5 s^ -1 and 4.7 x 10^ 6 s^ -1 .M^ -1 , respectively. The purified phytase exhibited broad specificity on a range of phosphorylated compounds, presenting activity on sodium phytate, p-NPP, 2- naphthylphosphate, 1- naphthyl- phosphate, ATP, phenyl-phosphate, glucose-6-phosphate, calcium phytate and other substrates. Enzymatic activity was slightly inhibited by Mg^ 2+ , Cd^ 2+ , K^ + and Ca^ 2+ , and it was drastically inhibited by F^ - . The enzyme displayed high thermostability, retaining more than 90% activity at 60 °C during 120 h and displayed a t 1/2 of 94.5 h and 6.2 h at 70 °C and 80 °C, respectively. The enzyme demonstrated strong resistance toward pepsin and trypsin, and it retained more than 90% residual activity for both enzymes after 1 h treatment. Additionally, the enzyme efficiently hydrolyzed phytate in live- stock feed, liberating 15.3 mmol phosphate/mL after 2.5 h of treatment. |
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Monteiro, Paulo S.Guimarães, Valéria M.Melo, Ricardo R. deRezende, Sebastião T. de2017-11-01T17:27:37Z2017-11-01T17:27:37Z2015-03-0116784405http://dx.doi.org/10.1590/S1517-838220120037http://www.locus.ufv.br/handle/123456789/12735An Aspergillus niger UFV-1 phytase was characterized and made available for industrial application. The enzyme was purified via ultrafiltration followed by acid precipitation, ion exchange and gel filtration chromatography. This protein exhibited a molecular mass of 161 kDa in gel filtration and 81 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), indicating that it may be a dimer. It presented an optimum temperature of 60 °C and optimum pH of 2.0. The K M for so- dium phytate hydrolysis was 30.9 mM, while the k cat and k cat /K M were 1.46 x10^ 5 s^ -1 and 4.7 x 10^ 6 s^ -1 .M^ -1 , respectively. The purified phytase exhibited broad specificity on a range of phosphorylated compounds, presenting activity on sodium phytate, p-NPP, 2- naphthylphosphate, 1- naphthyl- phosphate, ATP, phenyl-phosphate, glucose-6-phosphate, calcium phytate and other substrates. Enzymatic activity was slightly inhibited by Mg^ 2+ , Cd^ 2+ , K^ + and Ca^ 2+ , and it was drastically inhibited by F^ - . The enzyme displayed high thermostability, retaining more than 90% activity at 60 °C during 120 h and displayed a t 1/2 of 94.5 h and 6.2 h at 70 °C and 80 °C, respectively. The enzyme demonstrated strong resistance toward pepsin and trypsin, and it retained more than 90% residual activity for both enzymes after 1 h treatment. Additionally, the enzyme efficiently hydrolyzed phytate in live- stock feed, liberating 15.3 mmol phosphate/mL after 2.5 h of treatment.engBrazilian Journal of Microbiology46 (1), p. 251–260, Mar. 2015PhosphatasePhytic acidDephosphorylationIsolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistanceinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINAL1517-8382-bjm-46-01-0251.pdf1517-8382-bjm-46-01-0251.pdftexto completoapplication/pdf1104294https://locus.ufv.br//bitstream/123456789/12735/1/1517-8382-bjm-46-01-0251.pdf9bfa55167cbe31a53173861c3313dff6MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://locus.ufv.br//bitstream/123456789/12735/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52THUMBNAIL1517-8382-bjm-46-01-0251.pdf.jpg1517-8382-bjm-46-01-0251.pdf.jpgIM Thumbnailimage/jpeg5428https://locus.ufv.br//bitstream/123456789/12735/3/1517-8382-bjm-46-01-0251.pdf.jpg51e51d4f18d3039f3cb7799da386da75MD53123456789/127352022-06-23 11:02:48.781oai:locus.ufv.br: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Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452022-06-23T14:02:48LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.en.fl_str_mv |
Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance |
title |
Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance |
spellingShingle |
Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance Monteiro, Paulo S. Phosphatase Phytic acid Dephosphorylation |
title_short |
Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance |
title_full |
Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance |
title_fullStr |
Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance |
title_full_unstemmed |
Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance |
title_sort |
Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance |
author |
Monteiro, Paulo S. |
author_facet |
Monteiro, Paulo S. Guimarães, Valéria M. Melo, Ricardo R. de Rezende, Sebastião T. de |
author_role |
author |
author2 |
Guimarães, Valéria M. Melo, Ricardo R. de Rezende, Sebastião T. de |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Monteiro, Paulo S. Guimarães, Valéria M. Melo, Ricardo R. de Rezende, Sebastião T. de |
dc.subject.pt-BR.fl_str_mv |
Phosphatase Phytic acid Dephosphorylation |
topic |
Phosphatase Phytic acid Dephosphorylation |
description |
An Aspergillus niger UFV-1 phytase was characterized and made available for industrial application. The enzyme was purified via ultrafiltration followed by acid precipitation, ion exchange and gel filtration chromatography. This protein exhibited a molecular mass of 161 kDa in gel filtration and 81 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), indicating that it may be a dimer. It presented an optimum temperature of 60 °C and optimum pH of 2.0. The K M for so- dium phytate hydrolysis was 30.9 mM, while the k cat and k cat /K M were 1.46 x10^ 5 s^ -1 and 4.7 x 10^ 6 s^ -1 .M^ -1 , respectively. The purified phytase exhibited broad specificity on a range of phosphorylated compounds, presenting activity on sodium phytate, p-NPP, 2- naphthylphosphate, 1- naphthyl- phosphate, ATP, phenyl-phosphate, glucose-6-phosphate, calcium phytate and other substrates. Enzymatic activity was slightly inhibited by Mg^ 2+ , Cd^ 2+ , K^ + and Ca^ 2+ , and it was drastically inhibited by F^ - . The enzyme displayed high thermostability, retaining more than 90% activity at 60 °C during 120 h and displayed a t 1/2 of 94.5 h and 6.2 h at 70 °C and 80 °C, respectively. The enzyme demonstrated strong resistance toward pepsin and trypsin, and it retained more than 90% residual activity for both enzymes after 1 h treatment. Additionally, the enzyme efficiently hydrolyzed phytate in live- stock feed, liberating 15.3 mmol phosphate/mL after 2.5 h of treatment. |
publishDate |
2015 |
dc.date.issued.fl_str_mv |
2015-03-01 |
dc.date.accessioned.fl_str_mv |
2017-11-01T17:27:37Z |
dc.date.available.fl_str_mv |
2017-11-01T17:27:37Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1590/S1517-838220120037 http://www.locus.ufv.br/handle/123456789/12735 |
dc.identifier.issn.none.fl_str_mv |
16784405 |
identifier_str_mv |
16784405 |
url |
http://dx.doi.org/10.1590/S1517-838220120037 http://www.locus.ufv.br/handle/123456789/12735 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartofseries.pt-BR.fl_str_mv |
46 (1), p. 251–260, Mar. 2015 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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Brazilian Journal of Microbiology |
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Brazilian Journal of Microbiology |
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