Oleifera Protein Research Articles

Moringa oleifera protein isolates as novel wall materials in the encapsulation of limonene: Physicochemical properties and its stability

The purpose of this study was to investigate the efficiency of Moringa oleifera protein isolates (MPI) as a wall material for encapsulating limonene. MPI (10 % concentration) formed stable emulsions with a mean droplet size of 1.63±0.07 μm and viscosity of 3.54±0.10 mPa·s. Characterization of microcapsules revealed encapsulation efficiency (94.77±0.05 %), yield (56.50±5.07 %), aqueous solubility (21.00±0.92 %), water activity (0.46±0.01), moisture (2.09±0.42 %), a light color, indicating suitability for incorporation into food products. MPI better protected limonene from light and temperature compared with soy protein isolate (SPI), commonly used in encapsulation. The Korsmeyer-Peppas model best fit the release behavior of MPI-limonene microcapsules (r2=0.959), indicating that the release was controlled by relaxation of polymer chains and erosion. The release constant (k0) implied that MPI (2.303) had a faster release rate of limonene than SPI (1.997). The findings indicate that MPI is a potential wall material for limonene encapsulation, with promising physicochemical properties and improved oxidative stability.

Expression Analysis and Interaction Protein Screening of CoGI, the Key Factor in Photoperiod Regulation of Flowering in Camellia oleifera Abel

Photoperiod is a pivotal regulatory factor in the flowering of Camellia oleifera Abel. (C. oleifera). GIGANTEA (GI) serves as a pivotal regulator, not only orchestrating the intricate circadian rhythm but also governing photoperiod-dependent flowering. In order to explore the function of GI in C. oleifera (CoGI), we obtained a CoGI gene-coding sequence and analyzed a CoGI protein sequence using bioinformatics. Furthermore, we conducted a spatiotemporal expression analysis of CoGI. And a yeast two-hybridization assay was used to screen the interacting proteins of CoGI. Evolutionary analysis revealed high conservation of the CoGI protein, which clustered with the GI protein from Camellia sinensis (CsGI) on a common evolutionary branch. The expression of CoGI was different in each part, and a tissue expression analysis revealed that the relative expression level of the CoGI gene is highest in the leaves of C. oleifera, while it is at its lowest in the seed coats. Transgenic Arabidopsis thaliana (Arabidopsis) overexpressing CoGI exhibited early flowering under long-day conditions. In addition, the yeast two-hybrid library screening revealed interactions between seven C. oleifera proteins and CoGI: CoACR9, CoLAO, CoDExH12-like, CoIT1K-like, CoUPF0481, CoIDM3, and CoAt4g27190-like. The findings demonstrated that CoGI is crucial to C. oleifera’s flowering.

Functional Properties of Moringa Oleifera Protein Isolates as Influenced by Different Isolation Techniques, pH, and Ionic Strength

AbstractPlant-based proteins are increasingly being utilized as they are sustainable, economical, and can enhance the organoleptic properties of food products. This study investigated the effect of extraction and isolation techniques (alkaline extraction & isoelectric precipitation and salt extraction & micellization) on the functional properties ((solubility, water holding and oil absorption capacity (WHC/OAC), emulsifying activity and stability index (EAI/ESI), foaming capacity and stability (FC/FS), and gelation)) of Moringa oleifera protein isolates. The effect of ionic strength (0.2–1.0 M NaCl) and pH (2–10) on the functional properties were also investigated. Salt extraction & micellization produced an isolate with a high solubility at 0.8 M NaCl and pH 10 (98.1%, 91.8%), EAI at 0.6 M NaCl and pH 6 (56.2 m2/g, 77.0 m2/g), and ESI at 0.8 M NaCl and pH 4 (64.4 min, 243.5 min). Alkaline extraction & isoelectric precipitation resulted in an isolate with a high WHC at 0.6 M NaCl and pH 10 (5.8 g/g, 4.0 g/g), OAC (4.9 g/g), FC at 0.4 M NaCl and pH 2 (203.3%, 163.3%), and gelation at 0.2 M NaCl, 0.4 M NaCl, and pH 8 (20.0%). Both extraction methods can be applied in the valorization of M. oleifera seedcake. Micellized isolate could be used in food emulsions due to better emulsifying properties, while isoelectric isolate can be applied in meats and sauces, owing to better water holding and oil absorption capacity. M. oleifera protein isolates are promising plant-based proteins that can be used in food formulations or to replace animal-derived proteins in food.

Physiochemical and functional properties of Moringa oleifera protein obtained by isoelectric precipitation

The work investigated the effect of extraction pH (10 or 11) and precipitation pH (4 or 4.5) on the physicochemical and functional properties of Moringa oleifera protein. The physicochemical characteristics of Moringa leaf powder were analyzed using proximate quantification based on AOAC standard methods. The thermal, spectral, x-ray diffraction data were collected using the differential scanning calorimetric, Fourier transform infrared spectroscopy (FTIR), and wide-angle x-ray diffraction. The extraction at pH 10 and the precipitation process at pH 4.5 generated the highest protein content of 58.88%. Some of the functional properties of this pH combination were emulsion activity at 29.67%, emulsion stability at 50.45%, protein solubility at 32.36%, and water absorption capacity of 4.78 mL water/g. The thermal properties suggested the denaturation temperature of Moringa protein powder at 75°C, which was comparable to that previously reported plantbased protein isolate. FTIR and x-ray spectra argued the domination of amorphous protein fraction with a trace of lipid and carbohydrate. With these characteristics, protein has great potential to be used for food and drug applications.

Moringa oleifera protein isolates: In silico analysis of bioactivity, allergenicity, and toxicity, and application in plant-based burgers

Moringa oleifera seeds are excellent sources of protein. In this study, the potential bioactivity, allergenicity, and toxicity of M. oleifera protein isolates (MPI), and the effect of different concentrations of MPI (0%, 5%, 10%, and 15%) on the physicochemical properties and sensory acceptance of plant-based burgers (PBB) were investigated. A total of 100 peptides were identified through LC-MS/MS analysis. Subsequent in silico analyses unveiled their notable bioactivity potential and their potential to cause allergies while showing low levels of toxicity. The addition of MPI improved the nutritional value and cooking properties of PBB, evidenced by a 13% increment in protein content, a 3% increment in moisture retention, and an 11% less cooking loss. Furthermore, burgers containing 15% MPI had the lowest hardness (11.65 ± 0.33 N), cohesiveness (0.25 ± 0.01), springiness (0.42 ± 0.02 mm), and chewiness (1.22 ± 0.11 N × mm). MPI did not impair (p > 0.05) consumers’ sensory acceptance of the PBB. Overall, MPI is a promising protein source for preparing high-quality PBB.

Expression Analysis and Interaction Protein Screening of CoZTL in Camellia oleifera Abel

Camellia oleifera Abel., which produces fruits of high comprehensive utilization value, is an important woody oil tree in China. ZEITLUPE (ZTL) is a blue light receptor and clock component protein that is involved in various physiological and biochemical processes. However, the expression pattern and function of C. oleifera ZTL (CoZTL) remain unclear. In this study, the coding sequence of the CoZTL gene was isolated and the protein function was explored using bioinformatics and expression analyses and heterologous expression techniques. The results showed that the CoZTL protein was highly conserved during evolution and was on the same branch of the evolutionary tree as the ZTL proteins from Ipomoea nil and Nicotiana attenuata. CoZTL was mainly expressed in the fruit shells and stems of C. oleifera, and its expression level fluctuated greatly during flower bud development. Transgenic CoZTL-overexpressing Arabidopsis plants showed delayed flowering under long-day conditions as well as light-dependent promotion of hypocotyl elongation. Furthermore, yeast two-hybrid library screening revealed that seven C. oleifera proteins (CoAAT, Coβ-GAL, CoLAT52-like, CoCAR4-like, CoAO, CoUQCC1, and CoADF 2) interacted with CoZTL. Our results indicate that CoZTL plays an important role in C. oleifera flowering and hypocotyl growth.

The Crosslinking Mechanism of Camellia Oleifera Protein Adhesive with Amine Resins by 13C-NMR and ESI-MS

The Crosslinking Mechanism of Camellia Oleifera Protein Adhesive with Amine Resins by 13C-NMR and ESI-MS

Identification and Analysis of MADS-box, WRKY, NAC, and SBP-box Transcription Factor Families in Diospyros oleifera Cheng and Their Associations with Sex Differentiation

Transcription factors are crucial for plant growth and development. MADS-box, WRKY, NAC, and SBP-box, some of the most vital transcription factors, are not well identified in Diospyros spp., and their floral development and sex differentiation related functions remain unknown. We identified and analysed 53 MADS-box, 66 WRKY, 83 NAC, and 17 SBP-box transcription factors using the chromosomal D. oleifera genome. There were six DolSBPs identified as miR156 and miR157 targets. According to the multiple sequence alignments of Arabidopsis and D. oleifera proteins and their conserved domains and motifs, DolMADSs were divided into 23 type I and 30 type II. The DolWRKYs, DolNACs, and DolSBPs were divided into 7, 16, and 6 subgroups, respectively. It was found that one DolMADS, five DolWRKYs, one DolNAC, and four DolSBPs may promote unisexual female flowers development, while three DolMADSs, four DolWRKYs, and one DolNAC may enhance unisexual male flowers development. The functions among the different members of the same family may, thus, vary widely. The results will help to elucidate the evolution of the MADS-box, WRKY, NAC, and SBP-box genes families in D. oleifera and to determine their functions in flower development for Diospyros spp.

The effect of isolation techniques on the physicochemical properties of Moringa oleifera protein isolates

The effect of alkaline extraction/isoelectric precipitation and salt extraction/micellization on proximate composition, anti-nutritional content, amino acid composition and physicochemical properties of Moringa oleifera protein isolates were studied. Protein extractability was influenced by NaCl concentration, pH and temperature. The protein concentration of isoelectric and micellized isolates were 80% and 90%, respectively. Micellized isolates recorded higher yields (49%) than isoelectric isolates (15%). Both isolates were free of phytate, tannin, saponin and oxalate. Micellized isolates contained all essential amino acids except lysine and methionine which met the FAO/WHO recommendations for children; isoelectric isolates were deficient in methionine. Molecular weights of protein fractions were approximately 8.5–32.5 kDa. Micellization resulted in isolates with a lighter color, higher surface hydrophobicity (184) and higher transition enthalpy (109 J/g); isoelectric precipitation resulted in isolates with a higher denaturation temperature (190 °C). The results indicated that M. oleifera protein isolates could be a potential source of plant-based protein. The differences in physicochemical properties between the isolates indicate different functionalities and potentially different applications.

Effect of heat treatment on the structure, functional properties and composition of Moringa oleifera seed proteins

Knowledge on how food processing conditions and protein composition can modulate individual or food matrix protein functionality is crucial for designing new protein ingredients. In this regard, we investigated how heat treatment and protein composition influence physicochemical and functional properties of Moringa oleifera seed protein isolate. Results showed that changes in processing temperature induced modifications in the conformation affecting the hydrophobic core of proteins. Protein isolate was more soluble at room temperature whereas prolamin fraction presented high solubility at 70 °C. Glutelin showed higher emulsifying properties at all temperatures. Protein composition also significantly affected physicochemical and functional properties of protein isolate. Increasing soluble glutelin enhanced solubility while increasing albumin, globulin and glutelin decreased hydrophobicity of the isolate. Likewise, increasing soluble globulin improved emulsifying capacity, and emulsion stability of the isolate was negatively affected by increase in albumin and glutelin. These findings could enhance application of Moringa oleifera protein in food formulations.

Coagulation of turbid wastewater with an active component extracted from Moringa oleifera seeds

ABSTRACT The oil and carbohydrates contained in the Moringa oleifera seeds (MO) are two parameters that inhibit its coagulant potential. Oil extraction and protein purification are therefore necessary to improve its performance. The extraction process to recover the maximum oil yield was carried out using the solvent chloroform: ethanol C: E in a ratio (3:1). At a temperature of 100°C, the yield was higher than 44%. Purification was carried out by salting-in the protein with 1 M NaCl combined with ethanol precipitation and heat treatment. Analysis of the protein powder by the Kjeldahl method revealed 92% pure protein and 14% nitrogen; energy dispersive X-ray (EDX) analysis revealed high percentages of carbon (45%) and oxygen (39%). Scanning electron microscopy (SEM) showed a porous surface, and X-ray diffraction identified a crystalline phase of protein. The isoelectric point of Moringa oleifera protein (MOP) is in the range of pH 6.5–7.5 and its zeta potential varies between −19 mv and + 25.49 mV. It indicates a lower level of dispersion stability, implying a high capacity for protein agglomeration and flocculation. By reducing the zeta potential of water from −16 mv to −1.18 mv after treatment without modifying its pH, the Moringa protein has proven its ability to destabilise organic colloids. Moringa protein as a coagulat aid is effective for high turbidity wastewater with removal up to 86.78% and DOC removal up to 56.52%. The study reveals that coagulation with Moringa protein is a cost-effective alternative since it reduces the quantity of coagulant and time required for wastewater treatment.

Study on Camellia Oleifera Protein based Wood Adhesive by Epoxy Resin and Its Crosslinking Mechanism

Study on Camellia Oleifera Protein based Wood Adhesive by Epoxy Resin and Its Crosslinking Mechanism

Predicted of a heat shock cognate 70 kDa protein by PHYRE2 and SWISS-MODEL software from Elaeis guineensis and E. oleifera

Protein modeling was obtained from Elaeis guineensis and E. oleifera. The identification of heat shock cognate70 kDaboth very similar to coverage 90% analyzed by SWISS-MODEL. In addition, the data from similarity, identity, range, and Q-mean were distinct. According to the PHYRE2 analysis, we observed the longest residue presenting in E. guineensis (15-348). Wherein the 3D-ligand site prediction confirms that many amino acids were found (GLY, THR, TYR, LYS, GLU, ARG, and SER). Interestingly, the heat shock cognate 70 kDa protein in E. oleifera has a slight residue (15-18), and also have been three amino acids (GLY, THR, TYR). Appropriately, the heat shock cognate-like protein involved in different species, likewise playing a distinct response to kind the defense. The different homology images and templates were identified as different binding site predictions. These models can be used for supporting the E. guineensis and E. oleifera proteins modeling, mainly in the molecular study.

Antihypertensive activity of the ACE-renin inhibitory peptide derived from Moringa oleifera protein.

Moringa oleifera (MO) leaf is a potential plant protein resource with high nutritional and medicinal value. The study aims to investigate the hypotensive activity and stability of MO leaf peptides. MO leaf protein was extracted and then hydrolyzed with Alcalase to produce the MO leaf protein hydrolysate (MOPH). The MOPH was separated into peptide fractions with different molecular weights by membrane ultrafiltration. The MOPH and ultrafiltration fractions were evaluated for antihypertensive activity. Inhibition of the angiotensin-converting enzyme (84.71 ± 0.07%) and renin (43.72 ± 0.02%) was significantly higher for <1 kDa peptides when compared to other fractions. Oral administration of the <1 kDa component in spontaneously hypertensive rats positively lowers the blood pressure (∼17 mmHg). The <1 kDa component was isolated and purified subsequently; the final active component was identified by mass spectrometry and amino acid sequence analysis. Two highly active ACE (angiotensin-converting enzyme) and renin dual inhibitory peptides Leu-Gly-Phe-Phe (LGF) and Gly-Leu-Phe-Phe (GLFF) were obtained. The two peptides exhibited a good dual inhibitory activity of ACE and renin with IC50 values of LGF (0.29 ± 0.13 mM, 1.88 ± 0.08 mM) and GLFF (0.31 ± 0.04 mM, 2.80 ± 0.08 mM). Furthermore, in vivo models, LGF and GLFF significantly reduced the systolic blood pressure (19.4 mmHg; 18.2 mmHg) and diastolic blood pressure (12 mmHg; 13.8 mmHg) of SHRs (spontaneously hypertensive rats). The peptide transmembrane transport experiments and simulated gastrointestinal digestion experiments with LGF and GLFF showed that they can resist gastrointestinal digestion in a complete form. Thus, bioactive peptides from MO leaf may possess the potential to be used for treating hypertension in humans.

Biologically active Camellia oleifera protein nanoparticles for improving the tumor microenvironment and drug delivery.

It is important for antitumor drugs to accumulate at the tumor site and penetrate deeply to play a role in treatment. However, it is difficult for the drugs to reach the destination on account of the complex tumor microenvironment such as elevated tumor interstitial fluid pressure (IFP) and solid stress. Here, we report a type of nanocarrier composed entirely of Camellia oleifera protein (COP), which could lower tumor IFP and solid stress. Its physicochemical properties, cellular uptake, in vitro cytotoxicity and tumor perfusion, biodistribution, and in vivo antitumor efficiency were evaluated. It was found that COP NPs had good cellular uptake ability and cytocompatibility. When loading doxorubicin, COP NPs showed an in vitro concentration-dependent cytotoxicity. Importantly, the tumor IFP and solid stress were greatly reduced after injecting COP NPs into tumor-bearing mice, leading to more drug accumulating in the tumor and a longer survival time for tumor-bearing mice. Therefore, our study provided a new strategy to improve the tumor microenvironment and to achieve better antitumor efficiency.

Antibacterial action of functional silicon dioxide: an investigation of the attachment and separation of bacteria

ABSTRACTBactericidal proteins from the Moringa oleifera seed are reported to be suitable alternatives to conventional methods of bacterial reduction in water. In this study the cationic bactericidal M. oleifera proteins were isolated by attachment onto the surface of silicon dioxide. This functionalised SiO2(ƒ-SiO2) was then exposed to Escherichia coli and Micrococcus luteus to examine whether the ƒ-SiO2 could be used to inactivate the bacteria. The effect of the non-ionic surfactant dodecyl glucoside on the attachment of these bacteria to the ƒ-SiO2 was examined with the aim of developing a method of reusable bacterial inactivation. The primary result of this study was that the E. coli could be readily separated from the ƒ-SiO2, allowing the ƒ-SiO2 to be used for further bacterial inactivation. The regeneration of the ƒ-SiO2 was demonstrated using fluorescence microscopy on bacterial cells stained with propidium iodide, and zeta potential measurements. Future applications of this work include a reusable method of removing bacteria from contaminated water.

Effect of humic acids on the kaolin coagulation performance of Moringa oleifera proteins

Cationic proteins from Moringa oleifera seeds are of interest as sustainable coagulants for drinking water treatment in regions with poor access to potable water. They have been shown to reduce turbidity in natural water sources as well as in various model freshwaters. Prior research on kaolin suspension coagulation by M. oleifera proteins demonstrated that coagulation occurs by adsorption and charge neutralization. Natural organic matter may interfere with this mechanism by complexing with proteins or adsorbing to kaolin particles. Electrophoretic mobility measurements of M. oleifera proteins added to mixtures of kaolin and humic acid confirmed that the coagulation mechanism remains neutralization induced by adsorption. Coagulation micro-assays were used to determine the coagulation activity of M. oleifera proteins in United States Environmental Protection Agency model freshwaters (ionic strengths ranging from 1.15 to 9.17 mM) as a function of humic acid concentration. Model freshwaters contained 0.5 g/L suspended kaolin and 2.5 to 10 ppm humic acid. Increasing humic acid concentrations necessitated higher protein concentrations to achieve turbidity reduction. The range of effective coagulating protein doses broadened as humic acid concentration increased and as water hardness increased. Electrophoretic mobility and dynamic light scattering measurements showed evidence of humic acid adsorption to kaolin particles. These coagulation studies reveal the mechanistic roles of particulate and natural organic matter that will likely contribute to the development of methods to treat natural water using M. oleifera seed proteins.

Moringa oleifera Seed Protein Adsorption to Silica: Effects of Water Hardness, Fractionation, and Fatty Acid Extraction.

Motivated by the proposed use of cationic protein-modified sand for water filtration in developing nations, this study concerns the adsorption of Moringa oleifera seed proteins to silica surfaces. These proteins were prepared in model waters of varying hardness and underwent different levels of fractionation, including fatty acid extraction and cation exchange chromatography. Adsorption isotherms were measured by ellipsometry, and the zeta potentials of the resulting protein-decorated surfaces were measured by the rotating disk streaming potential method. The results indicate that the presence of fatty acids has little effect on the M. oleifera cationic protein adsorption isotherm. Adsorption from the unfractionated extract was indistinguishable from that of the cationic protein isolates at low concentrations but yielded significantly greater extents of adsorption at high concentrations. Adsorption isotherms for samples prepared in model hard and soft fresh waters were indistinguishable from each other over the measured bulk solution concentration range, but adsorption from hard or soft water was more extensive than adsorption from deionized water at moderate protein concentrations. Streaming potential measurements showed that adsorption reversed the net sign of the zeta potential of silica from negative to positive for all protein fractions and water hardness conditions at protein bulk concentrations as low as 0.03 μg/mL. This suggests that sands can be effectively modified with M. oleifera proteins using small amounts of seed extract under various local water hardness conditions. Finally, ellipsometry indicated that M. oleifera proteins adsorb irreversibly with respect to rinsing in these model fresh waters, suggesting that the modified sand would be stable on repeated use for water filtration. These studies may aid in the design of a simple, effective, and sustainable water purification device for developing nations.

Sticking particles to solid surfaces using Moringa oleifera proteins as a glue.

Experimental studies have been made to test the idea that seed proteins from Moringa oleifera which are novel, natural flocculating agents for many particles could be used to promote adhesion at planar interfaces and hence provide routes to useful nanostructures. The proteins bind irreversibly to silica interfaces. Surfaces that had been exposed to protein solutions and rinsed were then exposed to dispersions of sulfonated polystyrene latex. Atomic force microscopy was used to count particle density and identified that the sticking probability was close to 1. Measurements with a quartz crystal microbalance confirmed the adhesion and indicated that repeated exposures to solutions of Moringa seed protein and particles increased the coverage. Neutron reflectivity and scattering experiments indicate that particles bind as a monolayer. The various results show that the 2S albumin seed protein can be used to fix particles at interfaces and suggest routes for future developments in making active filters or improved interfaces for photonic devices.

A Chitin-binding Protein Purified from Moringa oleifera Seeds Presents Anticandidal Activity by Increasing Cell Membrane Permeability and Reactive Oxygen Species Production.

Candida species are opportunistic pathogens that infect immunocompromised and/or immunosuppressed patients, particularly in hospital facilities, that besides representing a significant threat to health increase the risk of mortality. Apart from echinocandins and triazoles, which are well tolerated, most of the antifungal drugs used for candidiasis treatment can cause side effects and lead to the development of resistant strains. A promising alternative to the conventional treatments is the use of plant proteins. M. oleifera Lam. is a plant with valuable medicinal properties, including antimicrobial activity. This work aimed to purify a chitin-binding protein from M. oleifera seeds and to evaluate its antifungal properties against Candida species. The purified protein, named Mo-CBP2, represented about 0.2% of the total seed protein and appeared as a single band on native PAGE. By mass spectrometry, Mo-CBP2 presented 13,309 Da. However, by SDS-PAGE, Mo-CBP2 migrated as a single band with an apparent molecular mass of 23,400 Da. Tricine-SDS-PAGE of Mo-CBP2 under reduced conditions revealed two protein bands with apparent molecular masses of 7,900 and 4,600 Da. Altogether, these results suggest that Mo-CBP2 exists in different oligomeric forms. Moreover, Mo-CBP2 is a basic glycoprotein (pI 10.9) with 4.1% (m/m) sugar and it did not display hemagglutinating and hemolytic activities upon rabbit and human erythrocytes. A comparative analysis of the sequence of triptic peptides from Mo-CBP2 in solution, after LC-ESI-MS/MS, revealed similarity with other M. oleifera proteins, as the 2S albumin Mo-CBP3 and flocculating proteins, and 2S albumins from different species. Mo-CBP2 possesses in vitro antifungal activity against Candida albicans, C. parapsilosis, C. krusei, and C. tropicalis, with MIC50 and MIC90 values ranging between 9.45–37.90 and 155.84–260.29 μM, respectively. In addition, Mo-CBP2 (18.90 μM) increased the cell membrane permeabilization and reactive oxygen species production in C. albicans and promoted degradation of circular plasmid DNA (pUC18) from Escherichia coli. The data presented in this study highlight the potential use of Mo-CBP2 as an anticandidal agent, based on its ability to inhibit Candida spp. growth with apparently low toxicity on mammalian cells.