Isolation and characterization of an aspartic protease from salpichroa origanifolia fruits
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 enz...
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| Formato: | Capítulo de libro |
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Bentham Science Publishers B.V.
2015
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| Acceso en línea: | Registro en Scopus Handle Registro en la Biblioteca Digital |
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| LEADER | 21967caa a22018737a 4500 | ||
|---|---|---|---|
| 001 | PAPER-13874 | ||
| 003 | AR-BaUEN | ||
| 005 | 20230518204418.0 | ||
| 008 | 170725s2015 xx ||||fo|||| 00| 0 eng|d | ||
| 024 | 7 | |2 scopus |a 2-s2.0-84931259807 | |
| 024 | 7 | |2 cas |a 1,10 phenanthroline, 3829-86-5, 66-71-7; aspartic proteinase, 78169-47-8; benzylsulfonyl fluoride, 329-98-6; casein, 9000-71-9; chymosin, 9001-98-3; edetic acid, 150-43-6, 60-00-4; hemoglobin, 9008-02-0; n [n (3 carboxyoxirane 2 carbonyl)leucyl]agmatine, 66701-25-5; pepstatin, 26305-03-3, 39324-30-6; insulin, 9004-10-8; Aspartic Acid Proteases; Insulin | |
| 040 | |a Scopus |b spa |c AR-BaUEN |d AR-BaUEN | ||
| 030 | |a PPELE | ||
| 100 | 1 | |a Rocha, G.F. | |
| 245 | 1 | 0 | |a Isolation and characterization of an aspartic protease from salpichroa origanifolia fruits |
| 260 | |b Bentham Science Publishers B.V. |c 2015 | ||
| 270 | 1 | 0 | |m Parisi, M.G.; Área de Química Biológica, Departamento de Ciencias Básicas, Universidad Nacional de Luján, Ruta 5 y Avenida Constitución, Argentina |
| 506 | |2 openaire |e Política editorial | ||
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| 520 | 3 | |a 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. © 2015 Bentham Science Publishers. |l eng | |
| 593 | |a Área de Química Biológica, Departamento de Ciencias Básicas, Universidad Nacional de Luján, Ruta 5 y Avenida Constitución, Luján, Buenos Aires, Argentina | ||
| 593 | |a Laboratorio de Investigación de Proteínas Vegetales, Departamento de Ciencias Biológicas, Universidad Nacional de la Plata, 47 y 115 s/N, C.C. 711, La Plata, Argentina | ||
| 593 | |a Instituto de Investigaciones Biológicas, Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Mar Del Plata, Funes 3250, Mar del Plata, Argentina | ||
| 593 | |a Consejo Nacional de Investigaciones Científicas y Tecnológicas, CONICET, Argentina | ||
| 690 | 1 | 0 | |a CASEINOLYTIC ACTIVITY |
| 690 | 1 | 0 | |a PEPTIDE MASS FINGERPRINTING |
| 690 | 1 | 0 | |a PLANT ASPARTIC PROTEASE |
| 690 | 1 | 0 | |a PURIFICATION |
| 690 | 1 | 0 | |a SALPICHROA ORIGANIFOLIA |
| 690 | 1 | 0 | |a SALPICHROIN |
| 690 | 1 | 0 | |a 1,10 PHENANTHROLINE |
| 690 | 1 | 0 | |a ASPARTIC PROTEINASE |
| 690 | 1 | 0 | |a BENZYLSULFONYL FLUORIDE |
| 690 | 1 | 0 | |a CASEIN |
| 690 | 1 | 0 | |a CHYMOSIN |
| 690 | 1 | 0 | |a EDETIC ACID |
| 690 | 1 | 0 | |a HEMOGLOBIN |
| 690 | 1 | 0 | |a N [N (3 CARBOXYOXIRANE 2 CARBONYL)LEUCYL]AGMATINE |
| 690 | 1 | 0 | |a PEPSTATIN |
| 690 | 1 | 0 | |a SYNTHETIC PEPTIDE |
| 690 | 1 | 0 | |a ASPARTIC PROTEINASE |
| 690 | 1 | 0 | |a INSULIN |
| 690 | 1 | 0 | |a AMINO ACID SEQUENCE |
| 690 | 1 | 0 | |a ANION EXCHANGE CHROMATOGRAPHY |
| 690 | 1 | 0 | |a ARTICLE |
| 690 | 1 | 0 | |a CONTROLLED STUDY |
| 690 | 1 | 0 | |a ENZYME KINETICS |
| 690 | 1 | 0 | |a FRUIT |
| 690 | 1 | 0 | |a MATRIX ASSISTED LASER DESORPTION IONIZATION TIME OF FLIGHT MASS SPECTROMETRY |
| 690 | 1 | 0 | |a MOLECULAR WEIGHT |
| 690 | 1 | 0 | |a NONHUMAN |
| 690 | 1 | 0 | |a POLYACRYLAMIDE GEL ELECTROPHORESIS |
| 690 | 1 | 0 | |a PRECIPITATION |
| 690 | 1 | 0 | |a PROTEIN ANALYSIS |
| 690 | 1 | 0 | |a PROTEIN CLEAVAGE |
| 690 | 1 | 0 | |a PROTEIN HYDROLYSIS |
| 690 | 1 | 0 | |a PROTEIN ISOLATION |
| 690 | 1 | 0 | |a PROTEIN PURIFICATION |
| 690 | 1 | 0 | |a SALPICHROA ORIGANIFOLIA |
| 690 | 1 | 0 | |a SOLANACEAE |
| 690 | 1 | 0 | |a CHEMISTRY |
| 690 | 1 | 0 | |a ISOLATION AND PURIFICATION |
| 690 | 1 | 0 | |a MOLECULAR GENETICS |
| 690 | 1 | 0 | |a SALPICHROA ORIGANIFOLIA |
| 690 | 1 | 0 | |a SOLANACEAE |
| 690 | 1 | 0 | |a AMINO ACID SEQUENCE |
| 690 | 1 | 0 | |a ASPARTIC ACID PROTEASES |
| 690 | 1 | 0 | |a FRUIT |
| 690 | 1 | 0 | |a INSULIN |
| 690 | 1 | 0 | |a MOLECULAR SEQUENCE DATA |
| 690 | 1 | 0 | |a SOLANACEAE |
| 650 | 1 | 7 | |2 spines |a PH |
| 700 | 1 | |a Obregón, W.D. | |
| 700 | 1 | |a Muñoz, F. | |
| 700 | 1 | |a Guevara, M.G. | |
| 700 | 1 | |a Fernández, G. | |
| 700 | 1 | |a Rosso, A.M. | |
| 700 | 1 | |a Parisi, M.G. | |
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| 962 | |a info:eu-repo/semantics/article |a info:ar-repo/semantics/artículo |b info:eu-repo/semantics/publishedVersion | ||
| 999 | |c 74827 | ||