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

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 th...

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Detalles Bibliográficos
Autores principales:	Lufrano, Daniela, Faro, Rosário, Castanheira, Pedro, Parisi, Gustavo Daniel, Veríssimo, Paula, Vairo Cavalli, Sandra Elizabeth, Simões, Isaura, Faro, Carlos
Formato:	Articulo
Lenguaje:	Inglés
Publicado:	2012
Materias:	Biología Cirsium vulgare Asteraceae Bull thistle Cirsin Procirsin Aspartic proteinase Pepsin-like protease Milk-clotting activity
Acceso en línea:	http://sedici.unlp.edu.ar/handle/10915/119358
Aporte de:	SEDICI (UNLP) de Universidad Nacional de La Plata

id	I19-R120-10915-119358
record_format	dspace
institution	Universidad Nacional de La Plata
institution_str	I-19
repository_str	R-120
collection	SEDICI (UNLP)
language	Inglés
topic	Biología Cirsium vulgare Asteraceae Bull thistle Cirsin Procirsin Aspartic proteinase Pepsin-like protease Milk-clotting activity
spellingShingle	Biología Cirsium vulgare Asteraceae Bull thistle Cirsin Procirsin Aspartic proteinase Pepsin-like protease Milk-clotting activity Lufrano, Daniela Faro, Rosário Castanheira, Pedro Parisi, Gustavo Daniel Veríssimo, Paula Vairo Cavalli, Sandra Elizabeth Simões, Isaura Faro, Carlos Molecular cloning and characterization of procirsin, an active aspartic protease precursor from <i>Cirsium vulgare</i> (Asteraceae)
topic_facet	Biología Cirsium vulgare Asteraceae Bull thistle Cirsin Procirsin Aspartic proteinase Pepsin-like protease Milk-clotting activity
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.
format	Articulo Articulo
author	Lufrano, Daniela Faro, Rosário Castanheira, Pedro Parisi, Gustavo Daniel Veríssimo, Paula Vairo Cavalli, Sandra Elizabeth Simões, Isaura Faro, Carlos
author_facet	Lufrano, Daniela Faro, Rosário Castanheira, Pedro Parisi, Gustavo Daniel Veríssimo, Paula Vairo Cavalli, Sandra Elizabeth Simões, Isaura Faro, Carlos
author_sort	Lufrano, Daniela
title	Molecular cloning and characterization of procirsin, an active aspartic protease precursor from <i>Cirsium vulgare</i> (Asteraceae)
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)
publishDate	2012
url	http://sedici.unlp.edu.ar/handle/10915/119358
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Molecular cloning and characterization of procirsin, an active aspartic protease precursor from <i>Cirsium vulgare</i> (Asteraceae)

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