Purification and characterization of cysteine protease from miswak Salvadora persica

Abstract

BackgroundGenerally, 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.ResultsPhysical 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.ConclusionsThe 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.

The Overexpression of Zea mays Strigolactone Receptor Gene D14 Enhances Drought Resistance in Arabidopsis thaliana L.

“…Concurrently, the variation in absorbance was recorded at a wavelength of 240 nm. A suspension was formulated containing 50 mM of potassium dihydrogen phosphate buffer, 120 mM of Met, 0.1 mM of EDTA, 700 µM of NBT, 60 µM of riboflavin, and 0.2 mL of enzyme extract, which was then used for the assessment of superoxide dismutase (SOD) activity [73].…”

Purification and Characterization of Alkaline Protease Isolated from Cotton (Gossypium hirsutum) Seeds

“…It was noted from the above-reported results as depicted in figure 9 that cotton seed protease prefers hydrolyzed peptones rather than other proteins. However, our finding is in variance with the reports of other researchers who investigated the substrate specificity of protease isolated from different plants such as cysteine protease from the fruit of Babi Kiwi exhibited high activity toward the synthetic substrate ZLeu-Arg-MCA and moderate activity toward Z-Val-Leu-LysMCA, Z-Val-Val-Arg-MCA, Z-Phe-Arg-MCA and Pro-PheArg-MCA, protease from miswak roots (Abdulaal, 2018)showed that the enzyme acted on casein, hemoglobin, egg albumin, gelatin and bovine serum albumin, three proteases from citrus sinensis fruit peel prefer to hydrolyzed gelatin than casein and bovine serum albumin and protease from seeds of Citrulluscolocynthis identified significantly high rates of hydrolysis for natural proteins such as hemoglobin, casein, gelatin and bovine serum albumin compared to modified proteins such as azoalbumin and azocasein (Khan, Khan, Shah, & Khan, 2016) .…”

A Novel Senescence-Specific Gene (ZmSAG39) Negatively Regulates Darkness and Drought Responses in Maize

“…The superoxide dismutase (SOD) activity was analyzed in 3 mL reaction mixture (50 mM potassium phosphate buffer (pH 7.0), 130mM Met, 0.1 mM EDTA, 750 µM NBT, 60 µM riboflavin and 0.2 mL of enzyme extract). One unit of SOD activity was defined as the quantity of enzyme required to induce 50% inhibition of the NBT reduction measured at 560 nm [52]. The ascorbate peroxidase (AXP) activity was analyzed in a 3 mL reaction mixture containing 50 mM potassium phosphate buffer (pH 7.0), 0.5 mM ASA, 6 mM H 2 O 2 and 50 µL of enzyme extract, and the changes in the absorbance at 290 nm were measured over a period of 30 s [53].…”