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dc.contributor.author
Rocha, Gabriela F.
dc.contributor.author
Obregon, Walter David
dc.contributor.author
Muñoz, Fernando Felipe
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Guevara, Maria Gabriela
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Fernandez, Graciela del Valle
dc.contributor.author
Rosso, Adriana M.
dc.contributor.author
Parisi, Mónica G.
dc.date.available
2017-02-20T21:22:50Z
dc.date.issued
2015-03
dc.identifier.citation
Rocha, Gabriela F. ; Obregon, Walter David; Muñoz, Fernando Felipe; Guevara, Maria Gabriela; Fernandez, Graciela del Valle; et al.; Isolation and characterization of an Aspartic Protease from Salpichroa origanifolia Fruits; Bentham Science Publishers; Protein And Peptide Letters; 22; 4; 3-2015; 379-390
dc.identifier.issn
0929-8665
dc.identifier.uri
http://hdl.handle.net/11336/13206
dc.description.abstract
This report describes the purification of an aspartic protease (salpichroin) from ripe fruits of Salpichroa origanifolia (Solanaceae) starting with precipitation using organic solvents and anionexchange chromatography with 32.1% recovery and 13.4-fold purification. SDS-PAGE and zymograms of this enzyme showed a single band corresponding to an apparent molecular mass of approximately 32 kDa. The biochemical and kinetic characterization of the pure enzyme showed an acidic behavior with an optimal pH value around 3.0–4.5 with hemoglobin and 5.5–6.0 with casein. Salpichroin activity was inhibited by pepstatin but not by phenylmethylsulfonyl fluoride, E-64, EDTA or 1,10-phenanthroline, thus suggesting an aspartic protease behavior. Salpichroin hydrolyzed natural substrates, such as casein and hemoglobin, with high specific activity. Kinetic studies conducted with the synthetic peptide H-Pro- Thr-Glu-Phe-p-(NO2)-Phe-Arg-Leu-OH showed lower affinity (Km 494 µM) than other representative aspartic proteases. By investigating the cleavage of oxidized insulin β-chain to establish the hydrolytic specificity of salpichroin, we found six cleavage sites on the substrate of peptide bonds similar to those of chymosin. MALDI-TOF/TOF-MS of the tryptic ingel digest of salpichroin showed that the isolated protease shared homologous sequences with other plant proteases of the A1 aspartic protease family. This is the first report concerning the isolation and biochemical characterization of an aspartic protease isolated from Salpichroa origanifolia fruits.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Bentham Science Publishers
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Caseinolytic Activity
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Peptide Mass Fingerprinting
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Plant Aspartic Protease
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Purification
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Salpichroa Origanifolia
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Salpichroin
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Bioquímica y Biología Molecular
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Ciencias Biológicas
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CIENCIAS NATURALES Y EXACTAS
dc.title
Isolation and characterization of an Aspartic Protease from Salpichroa origanifolia Fruits
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2017-02-13T20:24:33Z
dc.identifier.eissn
1875-5305
dc.journal.volume
22
dc.journal.number
4
dc.journal.pagination
379-390
dc.journal.pais
Estados Unidos
dc.description.fil
Fil: Rocha, Gabriela F. . Universidad Nacional de Luján. Departamento de Ciencias Básicas. Area de Química Biológica; Argentina
dc.description.fil
Fil: Obregon, Walter David. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigacion de Proteinas Vegetales; Argentina
dc.description.fil
Fil: Muñoz, Fernando Felipe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata; Argentina
dc.description.fil
Fil: Guevara, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata; Argentina
dc.description.fil
Fil: Fernandez, Graciela del Valle. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Area de Química Biológica; Argentina
dc.description.fil
Fil: Rosso, Adriana M. . Universidad Nacional de Luján. Departamento de Ciencias Básicas. Area de Química Biológica; Argentina
dc.description.fil
Fil: Parisi, Mónica G. . Universidad Nacional de Luján. Departamento de Ciencias Básicas. Area de Química Biológica; Argentina
dc.journal.title
Protein And Peptide Letters
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://www.eurekaselect.com/128937/article
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.2174/0929866522666150302111059
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