Trypsin purification using magnetic particles of azocasein-iron composite

Detalhes bibliográficos
Autor(a) principal: Alves, Maria Helena
Data de Publicação: 2017
Outros Autores: Nascimento, Gabriela Ayres, Cabrera, Mariana Paola, Silvério, Sara C., Gonçalves, Clarisse Salomé Nobre, Teixeira, J. A., Júnior, Luiz Bezerra de Carvalho
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/1822/44424
Resumo: This work presents an inexpensive, simple and fast procedure to purify trypsin based on affinity binding with ferromagnetic particles of azocasein composite (mAzo). Crude extract was obtained from intestines of fish Nile tilapia (Oreochromis niloticus) homogenized in buffer (01 g tissue/ml). This extract was exposed to 100mg of mAzo and washed to remove unbound proteins by magnetic field. Trypsin was leached off under high ionic strength (3 M NaCl). Preparation was achieved containing specific activity about 60 times higher than that of the crude extract. SDS-PAGE showed that the purified protein had molecular weight (24 kDa) in concordance with the literature for the Nile tilapia trypsin. The mAzo composite can be reused and applied to purify trypsin from other sources.
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spelling Trypsin purification using magnetic particles of azocasein-iron compositeEnzyme purificationProteasesFish visceraAzocasein iron oxidesScience & TechnologyThis work presents an inexpensive, simple and fast procedure to purify trypsin based on affinity binding with ferromagnetic particles of azocasein composite (mAzo). Crude extract was obtained from intestines of fish Nile tilapia (Oreochromis niloticus) homogenized in buffer (01 g tissue/ml). This extract was exposed to 100mg of mAzo and washed to remove unbound proteins by magnetic field. Trypsin was leached off under high ionic strength (3 M NaCl). Preparation was achieved containing specific activity about 60 times higher than that of the crude extract. SDS-PAGE showed that the purified protein had molecular weight (24 kDa) in concordance with the literature for the Nile tilapia trypsin. The mAzo composite can be reused and applied to purify trypsin from other sources.Authors are thankful to CNPq (grant 307273/2013-0) and CAPES (Scholarship to MHMEA) Brazil agencies as well as FCT (Scholarship to SICS and CN) Portugal Agencie.ElsevierUniversidade do MinhoAlves, Maria HelenaNascimento, Gabriela AyresCabrera, Mariana PaolaSilvério, Sara C.Gonçalves, Clarisse Salomé NobreTeixeira, J. A.Júnior, Luiz Bezerra de Carvalho2017-072017-07-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/44424engAlves, M.; Nascimento, Gabriela Ayres; Cabrera, Mariana Paola; Silvério, Sara C.; Nobre, Clarisse; Teixeira, J. A.; Júnior, Luiz Bezerra de Carvalho, Trypsin purification using magnetic particles of azocasein-iron composite. Food Chemistry, 226, 75-78, 20170308-814610.1016/j.foodchem.2016.12.09428254021http://www.elsevier.com/locate/issn/03088146info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:51:08Zoai:repositorium.sdum.uminho.pt:1822/44424Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:49:58.364175Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Trypsin purification using magnetic particles of azocasein-iron composite
title Trypsin purification using magnetic particles of azocasein-iron composite
spellingShingle Trypsin purification using magnetic particles of azocasein-iron composite
Alves, Maria Helena
Enzyme purification
Proteases
Fish viscera
Azocasein iron oxides
Science & Technology
title_short Trypsin purification using magnetic particles of azocasein-iron composite
title_full Trypsin purification using magnetic particles of azocasein-iron composite
title_fullStr Trypsin purification using magnetic particles of azocasein-iron composite
title_full_unstemmed Trypsin purification using magnetic particles of azocasein-iron composite
title_sort Trypsin purification using magnetic particles of azocasein-iron composite
author Alves, Maria Helena
author_facet Alves, Maria Helena
Nascimento, Gabriela Ayres
Cabrera, Mariana Paola
Silvério, Sara C.
Gonçalves, Clarisse Salomé Nobre
Teixeira, J. A.
Júnior, Luiz Bezerra de Carvalho
author_role author
author2 Nascimento, Gabriela Ayres
Cabrera, Mariana Paola
Silvério, Sara C.
Gonçalves, Clarisse Salomé Nobre
Teixeira, J. A.
Júnior, Luiz Bezerra de Carvalho
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Alves, Maria Helena
Nascimento, Gabriela Ayres
Cabrera, Mariana Paola
Silvério, Sara C.
Gonçalves, Clarisse Salomé Nobre
Teixeira, J. A.
Júnior, Luiz Bezerra de Carvalho
dc.subject.por.fl_str_mv Enzyme purification
Proteases
Fish viscera
Azocasein iron oxides
Science & Technology
topic Enzyme purification
Proteases
Fish viscera
Azocasein iron oxides
Science & Technology
description This work presents an inexpensive, simple and fast procedure to purify trypsin based on affinity binding with ferromagnetic particles of azocasein composite (mAzo). Crude extract was obtained from intestines of fish Nile tilapia (Oreochromis niloticus) homogenized in buffer (01 g tissue/ml). This extract was exposed to 100mg of mAzo and washed to remove unbound proteins by magnetic field. Trypsin was leached off under high ionic strength (3 M NaCl). Preparation was achieved containing specific activity about 60 times higher than that of the crude extract. SDS-PAGE showed that the purified protein had molecular weight (24 kDa) in concordance with the literature for the Nile tilapia trypsin. The mAzo composite can be reused and applied to purify trypsin from other sources.
publishDate 2017
dc.date.none.fl_str_mv 2017-07
2017-07-01T00:00:00Z
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://hdl.handle.net/1822/44424
url http://hdl.handle.net/1822/44424
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Alves, M.; Nascimento, Gabriela Ayres; Cabrera, Mariana Paola; Silvério, Sara C.; Nobre, Clarisse; Teixeira, J. A.; Júnior, Luiz Bezerra de Carvalho, Trypsin purification using magnetic particles of azocasein-iron composite. Food Chemistry, 226, 75-78, 2017
0308-8146
10.1016/j.foodchem.2016.12.094
28254021
http://www.elsevier.com/locate/issn/03088146
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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