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• W1418737564 abstract "SUMMARY. The presence of hydrolases in the latex of Araujia angustifolia (Asclepiadaceae), a climbing plant that grows in Argentina, has been studied. The crude extract (CE) obtained by differential centrifugation at 8000 and 15000 rpm of latex from A. angustifolia fruits, collected on 0.05 M citric-citrate buffer (pH 4.5) with 5 mM EDTA, exhibited several enzymatic activities. CE showed amydasic activity on L-pyroglutamyl-L-phenylalanyl-L-leucine-p-nitroanilide (PFLNA), proteolytic activity on 1% casein in TrisHCl pH 8.5, polygalacturonidase activity using 1% polygalacturonic acid in acetic acetate buffer pH 4.5, pectin methyl esterase on 1% pectin in citric phosphate pH 6.8 in the presence of 0.02M Cl 2Ca, and endosterolytic activity on p-nitrophenyl esters of N-α -carbobenzoxy-L-amino acids in Tris-HCl buffer pH 8.0. As proteolytic activity was the main hydrolytic activity observed, the proteolytic properties of CE were stablished. The proteases present in CE showed the characteristics of the cysteine proteases: optimum pH at alkaline range, isoelectric point (pI) higher than 8.0, and inhibition of activity by thiol blocking reagents, such as E-64 and iodoacetate. A remarkable thermal stability was also evident in the CE. Three proteases have been detected by IEF and zymogram in the CE and purified by FPLC affording three basic active fractions (araujiain aI, aII and aIII) with molecular masses about 23 kDa (SDS-PAGE). RESUMEN.“ Perfil Hidrolitico y Aislamiento de los Componentes Proteoliticos Presentes en el Latex de Frutos de Araujia angustifolia”. Se estudiaron las hidrolasas presentes en el l atex de Araujia angustifolia (Asclepiadaceae), enredadera que crece en Argentina. El extracto crudo (CE) obtenido por centrifugaci o n diferencial a 8000 y 15000 rpm del latex de frutos de A. angustifolia, recogido en buffer citrico-citrato 0,05 M (pH 4,5) con EDTA 5 mM, exhibio varias actividades enzimaticas: amidasica con L-piroglutamil-L-fenilalanil-L-leucina- p-nitroanilida (PFLNA) como sustrato; proteolitica sobre caseina al 1% en Tris-HCl pH 8,5; poligalacturonidasa usando acido poligalacturo nico al 1% en buffer ace tico-acetato pH 4,5; pectin metil esterasa ensayada sobre pectina al 1% en buffer citrico-fosfato pH 6,8, en presencia de Cl 2Ca 0,02M y endosterolitica empleando los e steres p-nitrofenilados de N-α -carbobenzoxi-L-aminoacidos en buffer Tris-HCl. pH 8,0. Dado que la actividad proteol itica fue la principal actividad hidrol itica observada, se determinaron las caracter isticas proteoliticas del CE, que contiene proteasas de tipo ciste inico: pH o ptimo alcalino, punto isoele ctrico (pI) superior a 8,0 y resultan inhibidas por reactivos bloqueantes de grupos ti o licos, tales como iodoacetato y E-64. El CE mostr o notable estabilidad t e rmica. Se detectaron tres proteasas por IEF y zimograma y luego fueron purificadas por FPLC. Las masas moleculares de las mismas fueron del orden de 23 kDa (SDS-PAGE). Las mismas fueron denominadas araujiaina aI, aII y aIII." @default.
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• W1418737564 date "2006-01-01" @default.
• W1418737564 modified "2023-09-27" @default.
• W1418737564 title "Hidrolytic Profile and isolation of the Proteolytic Components of Latex from Araujia angustifoliaFruits" @default.
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