Characterization of a Thermotolerant Phytase Produced by Rhizopus microsporus var. microsporus Biofilm on an Inert Support Using Sugarcane Bagasse as Carbon Source
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1007/s12010-016-2018-7 http://hdl.handle.net/11449/168416 |
Resumo: | The Rhizopus microsporus var. microsporus biofilm was able to produce increased levels of an extracellular thermotolerant phytase using polyethylene and viscose as an inert support in both modified NBRIP medium and modified Khanna medium containing sugarcane bagasse as the carbon source. The enzyme production was strictly regulated by the phosphorus content with optimal production at 0.5 mM of sodium phytate and KH2PO4. The extracellular phytase, RMPhy1, was purified 4.18-fold with 4.78 % recovery using DEAE-cellulose and CM-cellulose. A single protein band with a molecular mass of 35.4 kDa was obtained when the samples were subjected to 10 % SDS-PAGE. The optimum temperature for activity was 55 °C and the optimum pH was 4.5. R. microsporus var. microsporus phytase exhibited high stability at 30 and 40 °C with a half-life of 115 min at 60 °C. The enzyme activity increased in the presence of Ca2+ and was inhibited by Zn2+, arsenate, and sodium phosphate. Phytase demonstrated high substrate specificity for sodium phytate with Km = 0.72 mM and Vmax = 94.55 U/mg of protein and for p-NPP with Km = 0.04 mM and Vmax = 106.38 U/mg of protein. The enzyme also hydrolyzed ATP, AMPc, glucose 6-phosphate, glucose 1-phosphate, and UDPG. This is the first report on phytase characterization delivered with biofilm technology. The properties of the enzyme account for its high potential for use in biotechnology and the possibility of application in different industrial sectors as feed in the future. |
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Characterization of a Thermotolerant Phytase Produced by Rhizopus microsporus var. microsporus Biofilm on an Inert Support Using Sugarcane Bagasse as Carbon SourceEnzyme characterizationFungal biofilmPhytaseRhizopus microsporusThe Rhizopus microsporus var. microsporus biofilm was able to produce increased levels of an extracellular thermotolerant phytase using polyethylene and viscose as an inert support in both modified NBRIP medium and modified Khanna medium containing sugarcane bagasse as the carbon source. The enzyme production was strictly regulated by the phosphorus content with optimal production at 0.5 mM of sodium phytate and KH2PO4. The extracellular phytase, RMPhy1, was purified 4.18-fold with 4.78 % recovery using DEAE-cellulose and CM-cellulose. A single protein band with a molecular mass of 35.4 kDa was obtained when the samples were subjected to 10 % SDS-PAGE. The optimum temperature for activity was 55 °C and the optimum pH was 4.5. R. microsporus var. microsporus phytase exhibited high stability at 30 and 40 °C with a half-life of 115 min at 60 °C. The enzyme activity increased in the presence of Ca2+ and was inhibited by Zn2+, arsenate, and sodium phosphate. Phytase demonstrated high substrate specificity for sodium phytate with Km = 0.72 mM and Vmax = 94.55 U/mg of protein and for p-NPP with Km = 0.04 mM and Vmax = 106.38 U/mg of protein. The enzyme also hydrolyzed ATP, AMPc, glucose 6-phosphate, glucose 1-phosphate, and UDPG. This is the first report on phytase characterization delivered with biofilm technology. The properties of the enzyme account for its high potential for use in biotechnology and the possibility of application in different industrial sectors as feed in the future.Institute of Chemistry of Araraquara UNESP, Rua Francisco Degni s/nFaculty of Philosophy Sciences and Letters of Ribeirão Preto USP, Avenida Bandeirantes 3900, Monte AlegreInstitute of Chemistry of Araraquara UNESP, Rua Francisco Degni s/nUniversidade Estadual Paulista (Unesp)Universidade de São Paulo (USP)Sato, Vanessa Sayuri [UNESP]Jorge, João AtílioGuimarães, Luis Henrique Souza [UNESP]2018-12-11T16:41:10Z2018-12-11T16:41:10Z2016-06-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article610-624application/pdfhttp://dx.doi.org/10.1007/s12010-016-2018-7Applied Biochemistry and Biotechnology, v. 179, n. 4, p. 610-624, 2016.1559-02910273-2289http://hdl.handle.net/11449/16841610.1007/s12010-016-2018-72-s2.0-849591225332-s2.0-84959122533.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengApplied Biochemistry and Biotechnology0,571info:eu-repo/semantics/openAccess2024-01-27T06:52:27Zoai:repositorio.unesp.br:11449/168416Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-01-27T06:52:27Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Characterization of a Thermotolerant Phytase Produced by Rhizopus microsporus var. microsporus Biofilm on an Inert Support Using Sugarcane Bagasse as Carbon Source |
title |
Characterization of a Thermotolerant Phytase Produced by Rhizopus microsporus var. microsporus Biofilm on an Inert Support Using Sugarcane Bagasse as Carbon Source |
spellingShingle |
Characterization of a Thermotolerant Phytase Produced by Rhizopus microsporus var. microsporus Biofilm on an Inert Support Using Sugarcane Bagasse as Carbon Source Sato, Vanessa Sayuri [UNESP] Enzyme characterization Fungal biofilm Phytase Rhizopus microsporus |
title_short |
Characterization of a Thermotolerant Phytase Produced by Rhizopus microsporus var. microsporus Biofilm on an Inert Support Using Sugarcane Bagasse as Carbon Source |
title_full |
Characterization of a Thermotolerant Phytase Produced by Rhizopus microsporus var. microsporus Biofilm on an Inert Support Using Sugarcane Bagasse as Carbon Source |
title_fullStr |
Characterization of a Thermotolerant Phytase Produced by Rhizopus microsporus var. microsporus Biofilm on an Inert Support Using Sugarcane Bagasse as Carbon Source |
title_full_unstemmed |
Characterization of a Thermotolerant Phytase Produced by Rhizopus microsporus var. microsporus Biofilm on an Inert Support Using Sugarcane Bagasse as Carbon Source |
title_sort |
Characterization of a Thermotolerant Phytase Produced by Rhizopus microsporus var. microsporus Biofilm on an Inert Support Using Sugarcane Bagasse as Carbon Source |
author |
Sato, Vanessa Sayuri [UNESP] |
author_facet |
Sato, Vanessa Sayuri [UNESP] Jorge, João Atílio Guimarães, Luis Henrique Souza [UNESP] |
author_role |
author |
author2 |
Jorge, João Atílio Guimarães, Luis Henrique Souza [UNESP] |
author2_role |
author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) Universidade de São Paulo (USP) |
dc.contributor.author.fl_str_mv |
Sato, Vanessa Sayuri [UNESP] Jorge, João Atílio Guimarães, Luis Henrique Souza [UNESP] |
dc.subject.por.fl_str_mv |
Enzyme characterization Fungal biofilm Phytase Rhizopus microsporus |
topic |
Enzyme characterization Fungal biofilm Phytase Rhizopus microsporus |
description |
The Rhizopus microsporus var. microsporus biofilm was able to produce increased levels of an extracellular thermotolerant phytase using polyethylene and viscose as an inert support in both modified NBRIP medium and modified Khanna medium containing sugarcane bagasse as the carbon source. The enzyme production was strictly regulated by the phosphorus content with optimal production at 0.5 mM of sodium phytate and KH2PO4. The extracellular phytase, RMPhy1, was purified 4.18-fold with 4.78 % recovery using DEAE-cellulose and CM-cellulose. A single protein band with a molecular mass of 35.4 kDa was obtained when the samples were subjected to 10 % SDS-PAGE. The optimum temperature for activity was 55 °C and the optimum pH was 4.5. R. microsporus var. microsporus phytase exhibited high stability at 30 and 40 °C with a half-life of 115 min at 60 °C. The enzyme activity increased in the presence of Ca2+ and was inhibited by Zn2+, arsenate, and sodium phosphate. Phytase demonstrated high substrate specificity for sodium phytate with Km = 0.72 mM and Vmax = 94.55 U/mg of protein and for p-NPP with Km = 0.04 mM and Vmax = 106.38 U/mg of protein. The enzyme also hydrolyzed ATP, AMPc, glucose 6-phosphate, glucose 1-phosphate, and UDPG. This is the first report on phytase characterization delivered with biofilm technology. The properties of the enzyme account for its high potential for use in biotechnology and the possibility of application in different industrial sectors as feed in the future. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-06-01 2018-12-11T16:41:10Z 2018-12-11T16:41:10Z |
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/10.1007/s12010-016-2018-7 Applied Biochemistry and Biotechnology, v. 179, n. 4, p. 610-624, 2016. 1559-0291 0273-2289 http://hdl.handle.net/11449/168416 10.1007/s12010-016-2018-7 2-s2.0-84959122533 2-s2.0-84959122533.pdf |
url |
http://dx.doi.org/10.1007/s12010-016-2018-7 http://hdl.handle.net/11449/168416 |
identifier_str_mv |
Applied Biochemistry and Biotechnology, v. 179, n. 4, p. 610-624, 2016. 1559-0291 0273-2289 10.1007/s12010-016-2018-7 2-s2.0-84959122533 2-s2.0-84959122533.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Applied Biochemistry and Biotechnology 0,571 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
610-624 application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1799965737103654912 |