Molecular cloning and characterization of procirsin, an active aspartic protease precursor from <i>Cirsium vulgare</i> (Asteraceae)

Autores
Lufrano, Daniela; Faro, Rosário; Castanheira, Pedro; Parisi, Gustavo Daniel; Veríssimo, Paula; Vairo Cavalli, Sandra Elizabeth; Simões, Isaura; Faro, Carlos
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Typical aspartic proteinases from plants of the Astereaceae family like cardosins and cyprosins are wellknown milk-clotting enzymes. Their effectiveness in cheesemaking has encouraged several studies on other Astereaceae plant species for identification of new vegetable rennets. Here we report on the cloning, expression and characterization of a novel aspartic proteinase precursor from the flowers of Cirsium vulgare (Savi) Ten. The isolated cDNA encoded a protein product with 509 amino acids, termed cirsin, with the characteristic primary structure organization of plant typical aspartic proteinases. The pro form of cirsin was expressed in Escherichia coli and shown to be active without autocatalytically cleaving its pro domain. This contrasts with the acid-triggered autoactivation by pro-segment removal described for several recombinant plant typical aspartic proteinases. Recombinant procirsin displayed all typical proteolytic features of aspartic proteinases as optimum acidic pH, inhibition by pepstatin, cleavage between hydrophobic amino acids and strict dependence on two catalytic Asp residues for activity. Procirsin also displayed a high specificity towards j-casein and milk-clotting activity, suggesting it might be an effective vegetable rennet. The findings herein described provide additional evidences for the existence of different structural arrangements among plant typical aspartic proteinases.
Centro de Investigación de Proteínas Vegetales
Materia
Biología
Cirsium vulgare
Asteraceae
Bull thistle
Cirsin
Procirsin
Aspartic proteinase
Pepsin-like protease
Milk-clotting activity
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
SEDICI (UNLP)
Institución
Universidad Nacional de La Plata
OAI Identificador
oai:sedici.unlp.edu.ar:10915/119358
Acceder
id SEDICI_dc200c121fcb333e850fdc688fd212a8
oai_identifier_str oai:sedici.unlp.edu.ar:10915/119358
network_acronym_str SEDICI
repository_id_str 1329
network_name_str SEDICI (UNLP)
spelling Molecular cloning and characterization of procirsin, an active aspartic protease precursor from <i>Cirsium vulgare</i> (Asteraceae)Lufrano, DanielaFaro, RosárioCastanheira, PedroParisi, Gustavo DanielVeríssimo, PaulaVairo Cavalli, Sandra ElizabethSimões, IsauraFaro, CarlosBiologíaCirsium vulgareAsteraceaeBull thistleCirsinProcirsinAspartic proteinasePepsin-like proteaseMilk-clotting activityTypical aspartic proteinases from plants of the Astereaceae family like cardosins and cyprosins are wellknown milk-clotting enzymes. Their effectiveness in cheesemaking has encouraged several studies on other Astereaceae plant species for identification of new vegetable rennets. Here we report on the cloning, expression and characterization of a novel aspartic proteinase precursor from the flowers of Cirsium vulgare (Savi) Ten. The isolated cDNA encoded a protein product with 509 amino acids, termed cirsin, with the characteristic primary structure organization of plant typical aspartic proteinases. The pro form of cirsin was expressed in Escherichia coli and shown to be active without autocatalytically cleaving its pro domain. This contrasts with the acid-triggered autoactivation by pro-segment removal described for several recombinant plant typical aspartic proteinases. Recombinant procirsin displayed all typical proteolytic features of aspartic proteinases as optimum acidic pH, inhibition by pepstatin, cleavage between hydrophobic amino acids and strict dependence on two catalytic Asp residues for activity. Procirsin also displayed a high specificity towards j-casein and milk-clotting activity, suggesting it might be an effective vegetable rennet. The findings herein described provide additional evidences for the existence of different structural arrangements among plant typical aspartic proteinases.Centro de Investigación de Proteínas Vegetales2012info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdf7-18http://sedici.unlp.edu.ar/handle/10915/119358enginfo:eu-repo/semantics/altIdentifier/issn/0031-9422info:eu-repo/semantics/altIdentifier/doi/10.1016/j.phytochem.2012.05.028info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2025-09-29T11:28:10Zoai:sedici.unlp.edu.ar:10915/119358Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292025-09-29 11:28:10.602SEDICI (UNLP) - Universidad Nacional de La Platafalse
dc.title.none.fl_str_mv Molecular cloning and characterization of procirsin, an active aspartic protease precursor from <i>Cirsium vulgare</i> (Asteraceae)
title Molecular cloning and characterization of procirsin, an active aspartic protease precursor from <i>Cirsium vulgare</i> (Asteraceae)
spellingShingle Molecular cloning and characterization of procirsin, an active aspartic protease precursor from <i>Cirsium vulgare</i> (Asteraceae)
Lufrano, Daniela
Biología
Cirsium vulgare
Asteraceae
Bull thistle
Cirsin
Procirsin
Aspartic proteinase
Pepsin-like protease
Milk-clotting activity
title_short Molecular cloning and characterization of procirsin, an active aspartic protease precursor from <i>Cirsium vulgare</i> (Asteraceae)
title_full Molecular cloning and characterization of procirsin, an active aspartic protease precursor from <i>Cirsium vulgare</i> (Asteraceae)
title_fullStr Molecular cloning and characterization of procirsin, an active aspartic protease precursor from <i>Cirsium vulgare</i> (Asteraceae)
title_full_unstemmed Molecular cloning and characterization of procirsin, an active aspartic protease precursor from <i>Cirsium vulgare</i> (Asteraceae)
title_sort Molecular cloning and characterization of procirsin, an active aspartic protease precursor from <i>Cirsium vulgare</i> (Asteraceae)
dc.creator.none.fl_str_mv Lufrano, Daniela
Faro, Rosário
Castanheira, Pedro
Parisi, Gustavo Daniel
Veríssimo, Paula
Vairo Cavalli, Sandra Elizabeth
Simões, Isaura
Faro, Carlos
author Lufrano, Daniela
author_facet Lufrano, Daniela
Faro, Rosário
Castanheira, Pedro
Parisi, Gustavo Daniel
Veríssimo, Paula
Vairo Cavalli, Sandra Elizabeth
Simões, Isaura
Faro, Carlos
author_role author
author2 Faro, Rosário
Castanheira, Pedro
Parisi, Gustavo Daniel
Veríssimo, Paula
Vairo Cavalli, Sandra Elizabeth
Simões, Isaura
Faro, Carlos
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Biología
Cirsium vulgare
Asteraceae
Bull thistle
Cirsin
Procirsin
Aspartic proteinase
Pepsin-like protease
Milk-clotting activity
topic Biología
Cirsium vulgare
Asteraceae
Bull thistle
Cirsin
Procirsin
Aspartic proteinase
Pepsin-like protease
Milk-clotting activity
dc.description.none.fl_txt_mv Typical aspartic proteinases from plants of the Astereaceae family like cardosins and cyprosins are wellknown milk-clotting enzymes. Their effectiveness in cheesemaking has encouraged several studies on other Astereaceae plant species for identification of new vegetable rennets. Here we report on the cloning, expression and characterization of a novel aspartic proteinase precursor from the flowers of Cirsium vulgare (Savi) Ten. The isolated cDNA encoded a protein product with 509 amino acids, termed cirsin, with the characteristic primary structure organization of plant typical aspartic proteinases. The pro form of cirsin was expressed in Escherichia coli and shown to be active without autocatalytically cleaving its pro domain. This contrasts with the acid-triggered autoactivation by pro-segment removal described for several recombinant plant typical aspartic proteinases. Recombinant procirsin displayed all typical proteolytic features of aspartic proteinases as optimum acidic pH, inhibition by pepstatin, cleavage between hydrophobic amino acids and strict dependence on two catalytic Asp residues for activity. Procirsin also displayed a high specificity towards j-casein and milk-clotting activity, suggesting it might be an effective vegetable rennet. The findings herein described provide additional evidences for the existence of different structural arrangements among plant typical aspartic proteinases.
Centro de Investigación de Proteínas Vegetales
description Typical aspartic proteinases from plants of the Astereaceae family like cardosins and cyprosins are wellknown milk-clotting enzymes. Their effectiveness in cheesemaking has encouraged several studies on other Astereaceae plant species for identification of new vegetable rennets. Here we report on the cloning, expression and characterization of a novel aspartic proteinase precursor from the flowers of Cirsium vulgare (Savi) Ten. The isolated cDNA encoded a protein product with 509 amino acids, termed cirsin, with the characteristic primary structure organization of plant typical aspartic proteinases. The pro form of cirsin was expressed in Escherichia coli and shown to be active without autocatalytically cleaving its pro domain. This contrasts with the acid-triggered autoactivation by pro-segment removal described for several recombinant plant typical aspartic proteinases. Recombinant procirsin displayed all typical proteolytic features of aspartic proteinases as optimum acidic pH, inhibition by pepstatin, cleavage between hydrophobic amino acids and strict dependence on two catalytic Asp residues for activity. Procirsin also displayed a high specificity towards j-casein and milk-clotting activity, suggesting it might be an effective vegetable rennet. The findings herein described provide additional evidences for the existence of different structural arrangements among plant typical aspartic proteinases.
publishDate 2012
dc.date.none.fl_str_mv 2012
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Articulo
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://sedici.unlp.edu.ar/handle/10915/119358
url http://sedici.unlp.edu.ar/handle/10915/119358
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/issn/0031-9422
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.phytochem.2012.05.028
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
7-18
dc.source.none.fl_str_mv reponame:SEDICI (UNLP)
instname:Universidad Nacional de La Plata
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instname_str Universidad Nacional de La Plata
instacron_str UNLP
institution UNLP
repository.name.fl_str_mv SEDICI (UNLP) - Universidad Nacional de La Plata
repository.mail.fl_str_mv alira@sedici.unlp.edu.ar
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