Partial purification, immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillus
Autor(a) principal: | |
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Data de Publicação: | 2013 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://hdl.handle.net/11449/122670 |
Resumo: | Lipases have important applications in biotechnological processes, motivating us to produce, purify, immobilize and perform a biochemical characterization of the lipase from Rhizomucor pusillus. The fungus was cultivated by solid state fermentation producing lipolytic activity of about 0.5 U/mL(4U/g). A partial purification by gel filtration chromatography in Se-phacryl S-100 allowed obtaining a yield of about 85% and a purification factor of 5.7. Our results revealed that the purified enzyme is very stable with some significant differences in its properties when compared to crude extract. The crude enzyme extract has an optimum pH and temperature of 7.5 ° C and 40 ° C, respectively. After purification, a shift of the optimum pH from 7 to 8 was observed, as well as a rise in optimumtemperature to 60 ° C and an increase in stability. The enzyme was immobilized on CNBr-Agarose and Octyl-Agarose supports, having the highest immobilization yield of 94% in the second resin. The major advantage of immobilization in hydrophobic media such as Octyl is in its hyper activation, which in this case was over 200%, a very interesting finding. Another advantage of this type of immobilization is the possibility of using the derivatives in biotechnological applications, such as in oil enriched with omega-3 as the results obtained in this study display the hydrolysis of 40% EPA and 7% DHA from sardine oil, promising results compared to the literature. |
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Partial purification, immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillusEnzyme ImmobilizationSolid State FermentationPurificationn-3 Polyunsatured Fatty AcidsRhizomucor pusillusLipases have important applications in biotechnological processes, motivating us to produce, purify, immobilize and perform a biochemical characterization of the lipase from Rhizomucor pusillus. The fungus was cultivated by solid state fermentation producing lipolytic activity of about 0.5 U/mL(4U/g). A partial purification by gel filtration chromatography in Se-phacryl S-100 allowed obtaining a yield of about 85% and a purification factor of 5.7. Our results revealed that the purified enzyme is very stable with some significant differences in its properties when compared to crude extract. The crude enzyme extract has an optimum pH and temperature of 7.5 ° C and 40 ° C, respectively. After purification, a shift of the optimum pH from 7 to 8 was observed, as well as a rise in optimumtemperature to 60 ° C and an increase in stability. The enzyme was immobilized on CNBr-Agarose and Octyl-Agarose supports, having the highest immobilization yield of 94% in the second resin. The major advantage of immobilization in hydrophobic media such as Octyl is in its hyper activation, which in this case was over 200%, a very interesting finding. Another advantage of this type of immobilization is the possibility of using the derivatives in biotechnological applications, such as in oil enriched with omega-3 as the results obtained in this study display the hydrolysis of 40% EPA and 7% DHA from sardine oil, promising results compared to the literature.Universidade Estadual Paulista Júlio de Mesquita Filho, Departamento de Biologia, Instituto de Biociências Letras e Ciências Exatas de São José do Rio Preto, Sao Jose do Rio Preto, RUA CRISTOVAO COLOMBO, 2265, JARDIM NAZARETH, CEP 15054-000, SP, BrasilUniversidade Estadual Paulista Júlio de Mesquita Filho, Departamento de Biologia, Instituto de Biociências Letras e Ciências Exatas de São José do Rio Preto, Sao Jose do Rio Preto, RUA CRISTOVAO COLOMBO, 2265, JARDIM NAZARETH, CEP 15054-000, SP, BrasilDepartamento de Química e Ciências Ambientais, IBILCE UNESP - São José do Rio Preto.Universidade Estadual Paulista (Unesp)Institute of Catalysis and Petrochemistry, CSIC, Campus UAM, Cantoblanco, Madrid, SpainResearch Institute for Food Science, CIAL-CSIC, CalleNicolás Cabrera 9, Campus UAM, Madrid, SpainFerrarezzi, Ana LuciaPiveta, Daniele H.Bonilla-Rodriguez, Gustavo Orlando [UNESP]Silva, Roberto da [UNESP]Guisan, Jose ManuelGomes, Eleni [UNESP]Pessela, Benevides Costa2015-04-27T11:55:57Z2015-04-27T11:55:57Z2013info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article79-90application/pdfAdvances in Enzyme Research, v. 01, n.4, p. 79-90, 2013.2328-4846http://hdl.handle.net/11449/12267010.4236/aer.2013.14009ISSN2328-4846-2013-01-04-79-90.pdf942417568820654569552585886721307091241742851920Currículo Lattesreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengAdvances in Enzyme Researchinfo:eu-repo/semantics/openAccess2023-12-14T06:19:11Zoai:repositorio.unesp.br:11449/122670Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T20:18:00.707904Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Partial purification, immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillus |
title |
Partial purification, immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillus |
spellingShingle |
Partial purification, immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillus Ferrarezzi, Ana Lucia Enzyme Immobilization Solid State Fermentation Purification n-3 Polyunsatured Fatty Acids Rhizomucor pusillus |
title_short |
Partial purification, immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillus |
title_full |
Partial purification, immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillus |
title_fullStr |
Partial purification, immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillus |
title_full_unstemmed |
Partial purification, immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillus |
title_sort |
Partial purification, immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillus |
author |
Ferrarezzi, Ana Lucia |
author_facet |
Ferrarezzi, Ana Lucia Piveta, Daniele H. Bonilla-Rodriguez, Gustavo Orlando [UNESP] Silva, Roberto da [UNESP] Guisan, Jose Manuel Gomes, Eleni [UNESP] Pessela, Benevides Costa |
author_role |
author |
author2 |
Piveta, Daniele H. Bonilla-Rodriguez, Gustavo Orlando [UNESP] Silva, Roberto da [UNESP] Guisan, Jose Manuel Gomes, Eleni [UNESP] Pessela, Benevides Costa |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) Institute of Catalysis and Petrochemistry, CSIC, Campus UAM, Cantoblanco, Madrid, Spain Research Institute for Food Science, CIAL-CSIC, CalleNicolás Cabrera 9, Campus UAM, Madrid, Spain |
dc.contributor.author.fl_str_mv |
Ferrarezzi, Ana Lucia Piveta, Daniele H. Bonilla-Rodriguez, Gustavo Orlando [UNESP] Silva, Roberto da [UNESP] Guisan, Jose Manuel Gomes, Eleni [UNESP] Pessela, Benevides Costa |
dc.subject.por.fl_str_mv |
Enzyme Immobilization Solid State Fermentation Purification n-3 Polyunsatured Fatty Acids Rhizomucor pusillus |
topic |
Enzyme Immobilization Solid State Fermentation Purification n-3 Polyunsatured Fatty Acids Rhizomucor pusillus |
description |
Lipases have important applications in biotechnological processes, motivating us to produce, purify, immobilize and perform a biochemical characterization of the lipase from Rhizomucor pusillus. The fungus was cultivated by solid state fermentation producing lipolytic activity of about 0.5 U/mL(4U/g). A partial purification by gel filtration chromatography in Se-phacryl S-100 allowed obtaining a yield of about 85% and a purification factor of 5.7. Our results revealed that the purified enzyme is very stable with some significant differences in its properties when compared to crude extract. The crude enzyme extract has an optimum pH and temperature of 7.5 ° C and 40 ° C, respectively. After purification, a shift of the optimum pH from 7 to 8 was observed, as well as a rise in optimumtemperature to 60 ° C and an increase in stability. The enzyme was immobilized on CNBr-Agarose and Octyl-Agarose supports, having the highest immobilization yield of 94% in the second resin. The major advantage of immobilization in hydrophobic media such as Octyl is in its hyper activation, which in this case was over 200%, a very interesting finding. Another advantage of this type of immobilization is the possibility of using the derivatives in biotechnological applications, such as in oil enriched with omega-3 as the results obtained in this study display the hydrolysis of 40% EPA and 7% DHA from sardine oil, promising results compared to the literature. |
publishDate |
2013 |
dc.date.none.fl_str_mv |
2013 2015-04-27T11:55:57Z 2015-04-27T11:55:57Z |
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 |
Advances in Enzyme Research, v. 01, n.4, p. 79-90, 2013. 2328-4846 http://hdl.handle.net/11449/122670 10.4236/aer.2013.14009 ISSN2328-4846-2013-01-04-79-90.pdf 9424175688206545 6955258588672130 7091241742851920 |
identifier_str_mv |
Advances in Enzyme Research, v. 01, n.4, p. 79-90, 2013. 2328-4846 10.4236/aer.2013.14009 ISSN2328-4846-2013-01-04-79-90.pdf 9424175688206545 6955258588672130 7091241742851920 |
url |
http://hdl.handle.net/11449/122670 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Advances in Enzyme Research |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
79-90 application/pdf |
dc.source.none.fl_str_mv |
Currículo Lattes 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 |
|
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1808129185481228288 |