Vicilin has nutraceutical potential and different noteworthy medicative health-promoting biotic diversions, and it is remarkable against pathogenic microorganisms and insects. In this study, Vigna aconitifolia vicilin (VacV) has been identified and characterized from the seed of Vigna aconitifolia (Jacq.) Marechal (Moth beans). LC-MS/MS analysis of VacV provided seven random fragmented sequences comprising 238 residues, showing significant homology with already reported Vigna radiata vicilin (VraV). VacV was purified using ammonium sulfate precipitation (60%) followed by size exclusion chromatography on Hi-Load 16/60 Superdex 200 pg column and anion-exchange chromatography (Hi trap Q FF column). Purified VacV showed a major ~50 kDa band and multiple lower bands on 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under both reduced and non-reduced conditions. After all, a three-dimensional molecular structure of VacV was predicted, which showed β-sheeted molecular conformation similar to crystallographic structure of VraV. All Vicilins from V. aconitifolia and other plants were divided into six sub-groups by phylogenetic analysis, and VacV shared a high degree of similarity with vicilins of Vigna radiata, Pisum sativum, Lupinus albus, Cicer arietinum and Glycine max. Additionally, VacV (20 μg) has significant growth inhibition against different pathogenic bacteria along strong antifungal activity (50 μg). Likewise, VacV (3.0 mg) produced significant growth reduction in Rice Weevil Sitophilus oryzae larvae after 9 days compared with control. Furthermore, by using MMT assay, the cytotoxicity effect of VacV on the growth of HepG2 liver cancerous cells was tested. VacV showed cytotoxicity against the HepG-2 line and the acquired value was 180 µg after 48 h. Finally, we performed molecular docking against caspase-3 protein (PDB ID: 3DEI) for VacV bioactive receptor interface residues. Hence, our results reveal that VacV, has nutraceutical potential and moth beans can be used as a rich resource of functional foods.
Approach to fresh water is limited due to the growing world population and supply of clean drinking water is a global challenge. Pond water is widely used for drinking in developing countries and it has higher turbidity during rainy seasons. This purpose requires a large number of chemical coagulants that lead to a high cost of treatment. Therefore there is a need to find an economical and natural coagulant to treat turbid water. Coagulation activity is an ability of a liquid, changing to a solid or semi-solid state after sometime period of time. This study focuses on the isolation of coagulant proteins from Brassica nigra seeds responsible for water clarification. The coagulant protein (napin) was purified, quantified and identified. Napin protein was precipitated with 70% saturation of ammonium sulfate solution combined with dialysis and size-exclusion chromatography. SDS-PAGE showed that isolated napin has a molecular weight of 16 kDa. Protein quantification was done by using a NanoDrop spectrophotometer. The coagulation activity of the napin protein was measured against synthetic clay solution and turbid pond water. The coagulation assay results showed that napin protein from Brassica nigra has 85% and Moringa seeds extract has 87% coagulation activity against the synthetic clay solution after 120 min. However, the coagulation activity of napin protein against the turbid pond water was higher 85% compared to Moringa seed extract of 80%. The active coagulant napin protein would be helpful in establishing cheaper methods in scaling up the coagulant protein from the Brassica nigra seeds acting as a potential agent for water treatment.
In densely populated developing countries, water is limited, thus reservoirs are used and turbidity of the water rises during raining seasons. Because of this, a wide variety of chemical coagulants are required that raises the expense of treatment. The present research emphasizes mainly on using a plant-derived component as a natural coagulant and making ensure its supply is environmentally friendly and sustainable. Natural coagulants like carbohydrates, proteins exhibit the coagulation activity due to presence of charged particle and the suspended charge particles. This work focuses on the isolation and identification of natural coagulant proteins from Iberis umbellata L. seeds in order to improve the efficacy of turbidity removal and a coagulant protein chitin binding protein (CBP) has been purified and identified. Ammonium sulphate solution at 70% saturation was used to precipitate CBP through dialysis, and spin column chromatography was performed to purify it. Purified CBP has a molecular weight of around 14 kDa, determined by SDS-PAGE. The results showed that at varying pH levels, the coagulation activity of the seed extracts of I. umbellata and Moringa oleifera (Mo) was tested against turbid pond water and clay solution. At pH 7, the greatest reduction in turbidity was attained. After 120 min, the coagulation activity findings showed that Mo seed extract had 89% coagulation activity against the clay solution and CBP from I. umbellata had 87%. In contrast to M. oleifera (Mo) seed extract, which showed 82% coagulation activity, CBP showed a maximum of 83% coagulation activity against pond water. As a possible agent for water treatment, the active coagulant CBP protein would be useful in developing less expensive ways to scale up the coagulant protein from the seeds I. umbellata.
Photoinhibition and photooxidation of photosystem II (PSII) is one of the key damages caused by drought stress. The present study aimed to assess whether or not foliar application of copper oxide nanoparticles (CuO NPs) is effective in promoting PSII stability and activity in corn plants under drought stress. Three-week old plants of corn were subjected to drought stress and varying levels of CuO nano-particles (0, 25, 50, and 100 mM) of the rooting medium. In this study, the effects of foliar application of copper oxide nanoparticles were assessed on growth, plant water status, nutrient uptake and structural stability of photosystem II of drought stressed corn plants. Drought stress impeded overall growth of the corn plants by reducing leaf relative water content, water potential, chlorophyll a content, and accumulation of K+ in plant leaves and roots. Exogenous application of 25 mM CuO NPs significantly enhanced the growth of corn. Exogenous application of CuO NPs improved the dry biomass and length of roots of the corn plants under drought stress. Although the application of nano-particles did not change leaf photosynthetic pigments, relative water content, K+ accumulation, it enhanced the accumulation of K+ in roots. Drought stress did not affect the structural stability of PSII, but it reduced its activity (performance index, PIABS) due to changes in the reaction center density and the biochemical reaction efficiency or electron transport capability. Exogenous application of CuO NPs improved PIABS due to increase in active reaction center density and electron transport efficiency. However, 100 mM CuO NPs application caused PSII damage at the donor end and reduced active reaction center density in the corn plants under both normal and drought stress conditions. The present findings provide a baseline information that foliar application of CuO-nanoparticles in low concentrations can improve the growth of corn by improving accumulation of K+ and increasing PSII activity. Further research is required to explore its effect on cellular redox balance and activities of PSII and PSI, and optimum dose of CuO-nanoparticles for developing formulations for their commercial applications in farmers’ fields.