Partial purification, immobilization and preliminary biochemical characterization of lipases from Rhizomucor pusillus

Detalhes bibliográficos
Autor(a) principal: Ferrarezzi, Ana Lucia
Data de Publicação: 2013
Outros Autores: Piveta, Daniele H., Bonilla-Rodriguez, Gustavo Orlando [UNESP], Silva, Roberto da [UNESP], Guisan, Jose Manuel, Gomes, Eleni [UNESP], Pessela, Benevides Costa
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.
id UNSP_c649ef6cb2be28fb7a445a01b9a517fc
oai_identifier_str oai:repositorio.unesp.br:11449/122670
network_acronym_str UNSP
network_name_str Repositório Institucional da UNESP
repository_id_str 2946
spelling 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
_version_ 1808129185481228288