RESUMEN
BACKGROUND: Generally, proteases in medicinal plants had different therapeutic effects such as anti-inflammatory effect; modulate the immune response and inhibitory effect toward tumor growth. In this study, protease was purified and characterized from miswak roots, as medicinal plant and natural toothbrush. RESULTS: Physical and chemical characterization of cysteine protease P1 were studied such as pH optimum (6.5), optimum temperature (50 °C), thermal stability (50 °C) and Km (3.3 mg azocasein/ml). The enzyme digested some proteins in the order of caseine > haemoglobin > egg albumin >gelatin > bovine serum albumin. Hg2+ had strong inhibitory effect on enzyme activity compared with other metal ions. Kinetic of inhibition for determination the type of protease was studied. Iodoactamide and p-Hydroximercuribenzaoic acid (p-HMB) caused strong inhibitory effect on enzyme activity indicating the enzyme is cysteine protease. CONCLUSIONS: The biochemical characterization of this enzyme will be display the suitable conditions for using of this enzyme in toothpaste in the future and the enzyme may be used in other applications.
Asunto(s)
Proteasas de Cisteína/metabolismo , Proteínas de Plantas/metabolismo , Salvadoraceae/enzimología , Proteasas de Cisteína/química , Proteasas de Cisteína/genética , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/metabolismo , Estabilidad de Enzimas , Hidroximercuribenzoatos/química , Hidroximercuribenzoatos/metabolismo , Yodoacetamida/química , Yodoacetamida/metabolismo , Cinética , Mercurio/química , Mercurio/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/genética , Raíces de Plantas/enzimología , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/química , Proteínas Recombinantes/aislamiento & purificación , TemperaturaRESUMEN
Na2SeO3 supplementation in the dialysis medium could obviously prevent the dissociation of spectrin from the erythrocyte membranes. Such Se effect could be eliminated by pretreatment of erythrocyte membranes with a SH-blocking reagent, iodoacetamide(IAA) or addition of a SH-compound, dithio-threitol. The fluorescence intensity of erythrocyte membranes labelled with the fluorescent probe N-(3-pyrenyl)-maleimide decreased with increasing Na2SeO3 concentration used for pretreatment of ghosts. 31P-NMR spectra of erythrocyte membrane dialyzed in the presence or absence of Na2SeO3 concentration showed a difference in chemical shift anisotropy (delta sigma) between these two samples. These data suggest that the stabilization effect is based on changes in lipid-protein interaction and conformation of membrane skeletal components induced by reaction of their SH groups with Na2SeO3.