Immobilization and characterization of G51 keratinolytic enzymes with potential for wool processing

Autores
Iglesias, Martín Sebastián; Sequeiros, Cynthia; Islan, German Abel; Castro, Guillermo Raul; Olivera, Nelda Lila
Año de publicación
2018
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Bacillus sp. G51 produces extracellular keratinases with potential for shrink-proofing of wool. Keratinases are proteases with autolytic activity which are restringing their industrial application in free form. Immobilization could contribute to a better control of their catalytic activity. Our aim was to immobilize and characterize G51 extracellular enzymes by cross-linking of enzyme aggregates (CLEA). G51 culture supernatant was used for CLEA with glutaraldehyde as cross-linking agent. G51 enzyme units (EU)/glutaraldehyde ratio was optimized, obtaining the best recovery of the proteolytic activity with the lowest ratio tested (8.4% with 3.5 EU/mlglu25%). CLEA-G51 thermal stability was higher (91 and 71% of residual activity after 1 h at 50 and 60°C, respectively) than that of free enzymes (40 and 5% residual activity under the same conditions). After 4 month-storage at room temperature, the free and immobilized enzymes kept 20 and 80% of residual proteolytic activity, respectively. This improvement of storage stability suggests that immobilization could prevent G51-keratinase autolysis and loss of activity. More than 60% of the proteolytic activity was preserved in the 3rd use, and it gradually diminished to 30% after seven re-uses. CLEA-G51 enzymes retained its wool keratinolytic activity (0.06 EU/ml), which is essential for wool shrink-proofing. CLEA-G51 operational and storage advantages could be valuable for industrial applications. Particularly, increased molecular size of immobilized G51 keratinases could avoid their diffusion into the wool fiber, allowing wool treatments with higher enzyme concentrations and without excessive degradation.
Fil: Iglesias, Martín Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto Patagónico para el Estudio de los Ecosistemas Continentales; Argentina
Fil: Sequeiros, Cynthia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; Argentina
Fil: Islan, German Abel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina
Fil: Castro, Guillermo Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina
Fil: Olivera, Nelda Lila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto Patagónico para el Estudio de los Ecosistemas Continentales; Argentina
54th Annual Meeting Argentine Society for Biochemistry and Molecular Biology
Paraná
Argentina
Sociedad Argentina de Investigación en Bioquímica y Biología Molecular
Consejo Nacional de Investigaciones Científicas y Técnicas
Materia
BACILLUS
ENZYMES
KERATINOLYTIC
CLEA
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/227266
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/227266
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Immobilization and characterization of G51 keratinolytic enzymes with potential for wool processingIglesias, Martín SebastiánSequeiros, CynthiaIslan, German AbelCastro, Guillermo RaulOlivera, Nelda LilaBACILLUSENZYMESKERATINOLYTICCLEAhttps://purl.org/becyt/ford/2.9https://purl.org/becyt/ford/2Bacillus sp. G51 produces extracellular keratinases with potential for shrink-proofing of wool. Keratinases are proteases with autolytic activity which are restringing their industrial application in free form. Immobilization could contribute to a better control of their catalytic activity. Our aim was to immobilize and characterize G51 extracellular enzymes by cross-linking of enzyme aggregates (CLEA). G51 culture supernatant was used for CLEA with glutaraldehyde as cross-linking agent. G51 enzyme units (EU)/glutaraldehyde ratio was optimized, obtaining the best recovery of the proteolytic activity with the lowest ratio tested (8.4% with 3.5 EU/mlglu25%). CLEA-G51 thermal stability was higher (91 and 71% of residual activity after 1 h at 50 and 60°C, respectively) than that of free enzymes (40 and 5% residual activity under the same conditions). After 4 month-storage at room temperature, the free and immobilized enzymes kept 20 and 80% of residual proteolytic activity, respectively. This improvement of storage stability suggests that immobilization could prevent G51-keratinase autolysis and loss of activity. More than 60% of the proteolytic activity was preserved in the 3rd use, and it gradually diminished to 30% after seven re-uses. CLEA-G51 enzymes retained its wool keratinolytic activity (0.06 EU/ml), which is essential for wool shrink-proofing. CLEA-G51 operational and storage advantages could be valuable for industrial applications. Particularly, increased molecular size of immobilized G51 keratinases could avoid their diffusion into the wool fiber, allowing wool treatments with higher enzyme concentrations and without excessive degradation.Fil: Iglesias, Martín Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto Patagónico para el Estudio de los Ecosistemas Continentales; ArgentinaFil: Sequeiros, Cynthia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaFil: Islan, German Abel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; ArgentinaFil: Castro, Guillermo Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; ArgentinaFil: Olivera, Nelda Lila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto Patagónico para el Estudio de los Ecosistemas Continentales; Argentina54th Annual Meeting Argentine Society for Biochemistry and Molecular BiologyParanáArgentinaSociedad Argentina de Investigación en Bioquímica y Biología MolecularConsejo Nacional de Investigaciones Científicas y TécnicasTech Science Press2018info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/227266Immobilization and characterization of G51 keratinolytic enzymes with potential for wool processing; 54th Annual Meeting Argentine Society for Biochemistry and Molecular Biology; Paraná; Argentina; 2018; 70-701667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.techscience.com/biocell/v42nSuppl.4Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:53:38Zoai:ri.conicet.gov.ar:11336/227266instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-15 14:53:39.166CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Immobilization and characterization of G51 keratinolytic enzymes with potential for wool processing
title Immobilization and characterization of G51 keratinolytic enzymes with potential for wool processing
spellingShingle Immobilization and characterization of G51 keratinolytic enzymes with potential for wool processing
Iglesias, Martín Sebastián
BACILLUS
ENZYMES
KERATINOLYTIC
CLEA
title_short Immobilization and characterization of G51 keratinolytic enzymes with potential for wool processing
title_full Immobilization and characterization of G51 keratinolytic enzymes with potential for wool processing
title_fullStr Immobilization and characterization of G51 keratinolytic enzymes with potential for wool processing
title_full_unstemmed Immobilization and characterization of G51 keratinolytic enzymes with potential for wool processing
title_sort Immobilization and characterization of G51 keratinolytic enzymes with potential for wool processing
dc.creator.none.fl_str_mv Iglesias, Martín Sebastián
Sequeiros, Cynthia
Islan, German Abel
Castro, Guillermo Raul
Olivera, Nelda Lila
author Iglesias, Martín Sebastián
author_facet Iglesias, Martín Sebastián
Sequeiros, Cynthia
Islan, German Abel
Castro, Guillermo Raul
Olivera, Nelda Lila
author_role author
author2 Sequeiros, Cynthia
Islan, German Abel
Castro, Guillermo Raul
Olivera, Nelda Lila
author2_role author
author
author
author
dc.subject.none.fl_str_mv BACILLUS
ENZYMES
KERATINOLYTIC
CLEA
topic BACILLUS
ENZYMES
KERATINOLYTIC
CLEA
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.9
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv Bacillus sp. G51 produces extracellular keratinases with potential for shrink-proofing of wool. Keratinases are proteases with autolytic activity which are restringing their industrial application in free form. Immobilization could contribute to a better control of their catalytic activity. Our aim was to immobilize and characterize G51 extracellular enzymes by cross-linking of enzyme aggregates (CLEA). G51 culture supernatant was used for CLEA with glutaraldehyde as cross-linking agent. G51 enzyme units (EU)/glutaraldehyde ratio was optimized, obtaining the best recovery of the proteolytic activity with the lowest ratio tested (8.4% with 3.5 EU/mlglu25%). CLEA-G51 thermal stability was higher (91 and 71% of residual activity after 1 h at 50 and 60°C, respectively) than that of free enzymes (40 and 5% residual activity under the same conditions). After 4 month-storage at room temperature, the free and immobilized enzymes kept 20 and 80% of residual proteolytic activity, respectively. This improvement of storage stability suggests that immobilization could prevent G51-keratinase autolysis and loss of activity. More than 60% of the proteolytic activity was preserved in the 3rd use, and it gradually diminished to 30% after seven re-uses. CLEA-G51 enzymes retained its wool keratinolytic activity (0.06 EU/ml), which is essential for wool shrink-proofing. CLEA-G51 operational and storage advantages could be valuable for industrial applications. Particularly, increased molecular size of immobilized G51 keratinases could avoid their diffusion into the wool fiber, allowing wool treatments with higher enzyme concentrations and without excessive degradation.
Fil: Iglesias, Martín Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto Patagónico para el Estudio de los Ecosistemas Continentales; Argentina
Fil: Sequeiros, Cynthia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; Argentina
Fil: Islan, German Abel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina
Fil: Castro, Guillermo Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina
Fil: Olivera, Nelda Lila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto Patagónico para el Estudio de los Ecosistemas Continentales; Argentina
54th Annual Meeting Argentine Society for Biochemistry and Molecular Biology
Paraná
Argentina
Sociedad Argentina de Investigación en Bioquímica y Biología Molecular
Consejo Nacional de Investigaciones Científicas y Técnicas
description Bacillus sp. G51 produces extracellular keratinases with potential for shrink-proofing of wool. Keratinases are proteases with autolytic activity which are restringing their industrial application in free form. Immobilization could contribute to a better control of their catalytic activity. Our aim was to immobilize and characterize G51 extracellular enzymes by cross-linking of enzyme aggregates (CLEA). G51 culture supernatant was used for CLEA with glutaraldehyde as cross-linking agent. G51 enzyme units (EU)/glutaraldehyde ratio was optimized, obtaining the best recovery of the proteolytic activity with the lowest ratio tested (8.4% with 3.5 EU/mlglu25%). CLEA-G51 thermal stability was higher (91 and 71% of residual activity after 1 h at 50 and 60°C, respectively) than that of free enzymes (40 and 5% residual activity under the same conditions). After 4 month-storage at room temperature, the free and immobilized enzymes kept 20 and 80% of residual proteolytic activity, respectively. This improvement of storage stability suggests that immobilization could prevent G51-keratinase autolysis and loss of activity. More than 60% of the proteolytic activity was preserved in the 3rd use, and it gradually diminished to 30% after seven re-uses. CLEA-G51 enzymes retained its wool keratinolytic activity (0.06 EU/ml), which is essential for wool shrink-proofing. CLEA-G51 operational and storage advantages could be valuable for industrial applications. Particularly, increased molecular size of immobilized G51 keratinases could avoid their diffusion into the wool fiber, allowing wool treatments with higher enzyme concentrations and without excessive degradation.
publishDate 2018
dc.date.none.fl_str_mv 2018
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/227266
Immobilization and characterization of G51 keratinolytic enzymes with potential for wool processing; 54th Annual Meeting Argentine Society for Biochemistry and Molecular Biology; Paraná; Argentina; 2018; 70-70
1667-5746
CONICET Digital
CONICET
url http://hdl.handle.net/11336/227266
identifier_str_mv Immobilization and characterization of G51 keratinolytic enzymes with potential for wool processing; 54th Annual Meeting Argentine Society for Biochemistry and Molecular Biology; Paraná; Argentina; 2018; 70-70
1667-5746
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.techscience.com/biocell/v42nSuppl.4
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
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eu_rights_str_mv openAccess
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dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Tech Science Press
publisher.none.fl_str_mv Tech Science Press
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
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instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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