The mechanism of biocidal action of nano titania on Escherichia coli and Staphylococcus aureus has been evaluated by various biochemical techniques like lipid peroxidation, hydrolysis of orthonitrophenol β-D-galactopyranoside, estimation of protein-amino acid and bacterial nucleic acids leakage into solution, in addition to morphology studies by electron microscopy (TEM and SEM) and K(+) ion leakage by inductively coupled plasma optical emission spectrometry. The active anatase phase of nano titania has been synthesized by sol-gel and pulverization techniques to obtain particle sizes averaging around 11 nm. The nano semiconductor with a bandgap of 3.2 eV responds well to the UV source to liberate reactive oxygen species (ROS). Gram negative bacteria easily succumb to the ROS at a faster rate than gram-positive bacteria with an observable difference in the mode of attack. The use of analytical techniques revealed the release of peroxidized lipid (26 nmol mL(-1) ) and protein content (370 μg mL(-1)) with a K(+) ion concentration of 22 000 ppb on complete destruction of E. coli.
Pharmacological properties exhibited by latex of plants are due to various biologically active compounds including several proteolytic enzymes. Present study evaluates hemostatic potential of Tabernaemontana divaricata and Artocarpus altilis from Apocynaceae and Moraceae families respectively. The latex of these plants were initially subjected to dialysis and crude extracts were estimated for proteolytic activity using casein as the substrate. Mean caseinolytic activity for 100 μg of latex protein was found to be 56.16 ± 0.57 and 45 ± 0.3 U/h for T. divaricata and A. altilis respectively. Caseinolytic activity by both the plant extracts was higher than standard proteases, papain and trypsin. However the difference was significant (p < 0.05) with papain alone. Crude enzymes (CE) from both plants exhibited coagulant activity on human platelet poor plasma by recalcification time. A significant reduction in clotting time was exhibited by T. divaricata compared to A. altilis (p < 0.05). These results were further substantiated with fibrinogen agarose plate assay. Crude enzyme of both plants also hydrolyzed blood clot. Mean % of thrombolysis by T. divaricata was 80.75 ± 1.2 and that of A. altilis was 70.24 ± 1.52. Inhibition studies confirmed cysteine protease nature of CE. Comparative analysis revealed T. divaricata to be the best among the two for its hemostatic potential. This study scientifically validates the use of latex from these plants in the management of fresh cuts or wounds.
Introduction:Plant latex is a natural source of biologically active compounds and several hydrolytic enzymes responsible for their diverse health benefits. Recent past has witnessed substantial progress in understanding their supplementary industrial and pharmaceutical utility. Calotropis gigantea is one of the important latex producing plants belonging to asclepediaceae family with wide ethnopharmacological applications and is rich in proteolytic enzymes. Present study investigates hemostatic, milk clotting and blood stain removal potential of C. gigantea latex proteases.Materials and Methods:The protease activity of crude enzyme (CE), obtained by centrifugation followed by ammonium sulphate precipitation and dialysis, was assayed using casein as the substrate. Effect of pH, temperature and specific inhibitors on protease activity was determined. Native PAGE and in gel protease activity of CE was performed. Hemostatic (Fibrinogen polymerization, fibrinogen agarose plate and blood clot lysis assays), milk clotting and blood stain removal efficacies of CE were determined.Results:CE exhibited high caseinolytic activity. Enzyme activity was optimum at 37-50ºC and pH 8.0. Fibrinogen polymerization assay showed concentration dependent increase in turbidity indicating thrombin like activity which was further confirmed by fibrinogen agarose plate assays. Clot lysis assay indicated 92.41% thrombolysis by CE in 90 min. CE also revealed significantly high ratio of milk clotting to protease activity (Milk Clotting Index, MCI = 827.59 ± 1.52). Complete destaining of blood stained fabric was observed when incubated with 1% detergent incorporated with 0.1mg/ml CE. The study highlights and validates the compound application potential of latex cysteine proteases from C. gigantea.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.