Hydrolysis of α-lactalbumin by cardosin A immobilized on highly activated supports
Autor(a) principal: | |
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Data de Publicação: | 2003 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.14/6826 |
Resumo: | In the present research effort, production of derivatives of cardosin A (a plant protease) encompassing full stabilization of its dimeric structure has been achieved, via covalent, multi-subunit immobilization onto highly activated agarose-glutaraldehyde supports. Boiling such enzyme derivatives in the presence of sodium dodecyl sulfate and β-mercaptoethanol did not lead to leaching of enzyme, thus providing evidence for the effectiveness of the attachment procedure. Furthermore, the cardosin A derivatives prepared under optimal conditions presented ca. half the specific activity of the enzyme in soluble form, and were successfully employed at laboratory-scale trials to perform (selective) hydrolysis of α-lactalbumin (α-La), one of the major proteins in bovine whey. Hydrolysates of α-La were assayed for by the OPA method, as well as by FPLC, SDS–PAGE and HPLC. Thermal inactivation of the immobilized cardosin A was also assessed at 40, 50 and 55 °C; at these temperatures, no thermal denaturation took place during incubation for 48 h. The highest degree of hydrolysis was attained by 5 h reaction, at 55 °C and pH 5.2. SDS–PAGE of α-La hydrolysates displayed bands corresponding to low molecular weight peptides. Our results suggest that cardosin A in immobilized form is a good candidate to bring about proteolysis in the dairy industry, namely in whey processing. |
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Hydrolysis of α-lactalbumin by cardosin A immobilized on highly activated supportsEnzymeProteaseAgaroseAttachmentStructural stabilizationDairy foodsα-LactalbuminCardosinIn the present research effort, production of derivatives of cardosin A (a plant protease) encompassing full stabilization of its dimeric structure has been achieved, via covalent, multi-subunit immobilization onto highly activated agarose-glutaraldehyde supports. Boiling such enzyme derivatives in the presence of sodium dodecyl sulfate and β-mercaptoethanol did not lead to leaching of enzyme, thus providing evidence for the effectiveness of the attachment procedure. Furthermore, the cardosin A derivatives prepared under optimal conditions presented ca. half the specific activity of the enzyme in soluble form, and were successfully employed at laboratory-scale trials to perform (selective) hydrolysis of α-lactalbumin (α-La), one of the major proteins in bovine whey. Hydrolysates of α-La were assayed for by the OPA method, as well as by FPLC, SDS–PAGE and HPLC. Thermal inactivation of the immobilized cardosin A was also assessed at 40, 50 and 55 °C; at these temperatures, no thermal denaturation took place during incubation for 48 h. The highest degree of hydrolysis was attained by 5 h reaction, at 55 °C and pH 5.2. SDS–PAGE of α-La hydrolysates displayed bands corresponding to low molecular weight peptides. Our results suggest that cardosin A in immobilized form is a good candidate to bring about proteolysis in the dairy industry, namely in whey processing.ElsevierVeritati - Repositório Institucional da Universidade Católica PortuguesaBarros, Rui M.Extremina, Clara I.Gonçalves, Inês C.Braga, Beatriz O.Balcão, Victor M.Malcata, F. Xavier2011-10-22T15:46:19Z20032003-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.14/6826engBARROS, Rui M... [et al.] - Hydrolysis of α-lactalbumin by cardosin A immobilized on highly activated supports. Enzyme and Microbial Technology. ISSN: 0141-0229. Vol. 33, n.º 7 (2003) p. 908-91610.1016/S0141-0229(03)00244-8info: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:RCAAP2023-07-12T17:10:32Zoai:repositorio.ucp.pt:10400.14/6826Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:05:47.965855Repositório 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 |
Hydrolysis of α-lactalbumin by cardosin A immobilized on highly activated supports |
title |
Hydrolysis of α-lactalbumin by cardosin A immobilized on highly activated supports |
spellingShingle |
Hydrolysis of α-lactalbumin by cardosin A immobilized on highly activated supports Barros, Rui M. Enzyme Protease Agarose Attachment Structural stabilization Dairy foods α-Lactalbumin Cardosin |
title_short |
Hydrolysis of α-lactalbumin by cardosin A immobilized on highly activated supports |
title_full |
Hydrolysis of α-lactalbumin by cardosin A immobilized on highly activated supports |
title_fullStr |
Hydrolysis of α-lactalbumin by cardosin A immobilized on highly activated supports |
title_full_unstemmed |
Hydrolysis of α-lactalbumin by cardosin A immobilized on highly activated supports |
title_sort |
Hydrolysis of α-lactalbumin by cardosin A immobilized on highly activated supports |
author |
Barros, Rui M. |
author_facet |
Barros, Rui M. Extremina, Clara I. Gonçalves, Inês C. Braga, Beatriz O. Balcão, Victor M. Malcata, F. Xavier |
author_role |
author |
author2 |
Extremina, Clara I. Gonçalves, Inês C. Braga, Beatriz O. Balcão, Victor M. Malcata, F. Xavier |
author2_role |
author author author author author |
dc.contributor.none.fl_str_mv |
Veritati - Repositório Institucional da Universidade Católica Portuguesa |
dc.contributor.author.fl_str_mv |
Barros, Rui M. Extremina, Clara I. Gonçalves, Inês C. Braga, Beatriz O. Balcão, Victor M. Malcata, F. Xavier |
dc.subject.por.fl_str_mv |
Enzyme Protease Agarose Attachment Structural stabilization Dairy foods α-Lactalbumin Cardosin |
topic |
Enzyme Protease Agarose Attachment Structural stabilization Dairy foods α-Lactalbumin Cardosin |
description |
In the present research effort, production of derivatives of cardosin A (a plant protease) encompassing full stabilization of its dimeric structure has been achieved, via covalent, multi-subunit immobilization onto highly activated agarose-glutaraldehyde supports. Boiling such enzyme derivatives in the presence of sodium dodecyl sulfate and β-mercaptoethanol did not lead to leaching of enzyme, thus providing evidence for the effectiveness of the attachment procedure. Furthermore, the cardosin A derivatives prepared under optimal conditions presented ca. half the specific activity of the enzyme in soluble form, and were successfully employed at laboratory-scale trials to perform (selective) hydrolysis of α-lactalbumin (α-La), one of the major proteins in bovine whey. Hydrolysates of α-La were assayed for by the OPA method, as well as by FPLC, SDS–PAGE and HPLC. Thermal inactivation of the immobilized cardosin A was also assessed at 40, 50 and 55 °C; at these temperatures, no thermal denaturation took place during incubation for 48 h. The highest degree of hydrolysis was attained by 5 h reaction, at 55 °C and pH 5.2. SDS–PAGE of α-La hydrolysates displayed bands corresponding to low molecular weight peptides. Our results suggest that cardosin A in immobilized form is a good candidate to bring about proteolysis in the dairy industry, namely in whey processing. |
publishDate |
2003 |
dc.date.none.fl_str_mv |
2003 2003-01-01T00:00:00Z 2011-10-22T15:46:19Z |
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/10400.14/6826 |
url |
http://hdl.handle.net/10400.14/6826 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
BARROS, Rui M... [et al.] - Hydrolysis of α-lactalbumin by cardosin A immobilized on highly activated supports. Enzyme and Microbial Technology. ISSN: 0141-0229. Vol. 33, n.º 7 (2003) p. 908-916 10.1016/S0141-0229(03)00244-8 |
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 |
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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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1799131723463655424 |