Mutagens from roasted seeds of Moringa oleifera

A number of biosynthetically and chemically related compounds were isolated from the roasted seeds of Moringa oleifera. The micronucleus test, an in vivo method, using albino mice as the test system, was used for monitoring the mutagenicity of the isolated compounds. Structure-activity correlation studies showed that 4(alpha-L-rhamnosyloxy)phenylacetonitrile, 4-hydroxyphenylacetontrile, and 4-hydroxyphenyl-acetamide exhibited mutagenic activity.     

Flocculent activity of a recombinant protein from Moringa oleifera Lam. seeds

Seeds of the tropical tree Moringa oleifera contain small storage proteins able to flocculate particles in suspension in water. The cDNA encoding one of these flocculent proteins, MO(2.1), was cloned and the recombinant protein was expressed in Escherichia coli. The flocculent activity of the purified recombinant MO(2.1)was assayed on clays and bacteria using light and confocal microscopy and GFP-overexpressing bacteria. We show that MO(2.1)is able to aggregate montmorillonite clay particles as well as gram-positive and gram-negative bacteria. We discuss the use of recombinant proteins to study flocculating properties and improve water purification processes.     

Purification of a chitin-binding protein from Moringa oleifera seeds with potential to relieve pain and inflammation

Moringa oleifera Lam. is a perennial multipurpose tree that has been successfully used in folk medicine to cure several inflammatory processes. The aim of this study was to purify and characterize a chitin-binding protein from Moringa oleifera seeds, named Mo-CBP4, and evaluate its antinociceptive and anti-inflammatory effects in vivo. The protein was purified by affinity chromatography on chitin followed by ion exchange chromatography. Acetic acid-induced abdominal constrictions assay was used for the antinociceptive and anti-inflammatory activity assessments. Mo-CBP4 is a glycoprotein (2.9% neutral carbohydrate) composed of two protein subunits with apparent molecular masses of 28 and 18 kDa (9 kDa in the presence of reducing agent). The intraperitoneal injection of Mo-CBP4 (3.5 and 10 mg/kg) into mice 30 min before acetic acid administration potently and significantly reduced the occurrence of abdominal writhing in a dose dependent manner by 44.7% and 100%, respectively. In addition, the oral administration of the protein (10 mg/kg) resulted in 18% and 52.8% reductions in abdominal writhing when given 30 and 60 min prior to acetic acid administration, respectively. Mo-CBP4, when administered by intraperitoneal route, also caused a significant and dose-dependent inhibition of peritoneal capillary permeability induced by acid acetic and significantly inhibited leukocyte accumulation in the peritoneal cavity. In conclusion, this pioneering study describes that the chitin-binding protein Mo-CBP4, from M. oleifera seeds, exhibits anti-inflammatory and antinociceptive properties and scientifically supports the use of this multipurpose tree in folk medicine.     

Biosorption of trivalent and hexavalent chromium from aqueous systems using shelled Moringa oleifera seeds

Abstract

The present study explores the sorption properties of shelled Moringa oleifera seeds (SMOS) for removal of two environmentally important oxidation states of chromium (trivalent and hexavalent) from an aqueous system on the laboratory scale. Sorption studies reveal the optimum conditions for the removal of 81.02%; Cr (III) and 88.15% Cr (VI) as follows: biomass dosage (4.0 g), metal concentration [25mg/L for Cr (III); 50mg/L for Cr (VI)], contact time (40 minutes) at pH 6.5 and 2.5 respectively. The adsorption data were found to fit well both the Freundlich and Langmuir isotherms. Characterization of the seed powder by FTIR showed the clear presence of amino acid moieties having both positively charged amino and negatively charged carboxylic groups and confirmed that biosorption involves amino acid-chromium interactions. SEM studies of native and exhausted [Cr(III) and Cr(VI)] treated SMOS revealed large spherical clusters having a pore area of 8.66 µm² in the case of native SMOS while dense agglomerated etched dendrite type morphology have a pore area of 0.80 µm² in Cr (III) and 0.78 µm² in Cr (VI) treated SMOS The spent biosorbent was regenerated and found to be effectively reusable for four cycles.     

An antitumor promoter from Moringa oleifera Lam

In the course of studies on the isolation of bioactive compounds from Philippine plants, the seeds of Moringa oleifera Lam. were examined and from the ethanol extract were isolated the new O-ethyl-4-(alpha-L-rhamnosyloxy)benzyl carbamate (1) together with seven known compounds, 4(alpha-L-rhamnosyloxy)-benzyl isothiocyanate (2), niazimicin (3), niazirin (4), beta-sitosterol (5), glycerol-1-(9-octadecanoate) (6), 3-O-(6'-O-oleoyl-beta-D-glucopyranosyl)-beta-sitosterol (7), and beta-sitosterol-3-O-beta-D-glucopyranoside (8). Four of the isolates (2, 3, 7, and 8), which were obtained in relatively good yields, were tested for their potential antitumor promoting activity using an in vitro assay which tested their inhibitory effects on Epstein-Barr virus-early antigen (EBV-EA) activation in Raji cells induced by the tumor promoter, 12-O-tetradecanoyl-phorbol-13-acetate (TPA). All the tested compounds showed inhibitory activity against EBV-EA activation, with compounds 2, 3 and 8 having shown very significant activities. Based on the in vitro results, niazimicin (3) was further subjected to in vivo test and found to have potent antitumor promoting activity in the two-stage carcinogenesis in mouse skin using 7,12-dimethylbenz(a)anthracene (DMBA) as initiator and TPA as tumor promoter. From these results, niazimicin (3) is proposed to be a potent chemo-preventive agent in chemical carcinogenesis.     

Effects of storage conditions of Moringa oleifera seeds on its performance in coagulation

Moringa oleifera is a plant whose seeds have coagulation properties for treating water and wastewater. In this study the coagulation efficiency of Moringa oleifera kept in different storage conditions were studied. The Moringa oleifera seeds were stored at different conditions and durations; open container and closed container at room temperature (28 degrees C) and refrigerator (3 degrees C) for durations of 1, 3 and 5 months. Comparison between turbidity removal efficiency of Moringa oleifera kept in refrigerator and room temperature revealed that there was no significant difference between them. The Moringa oleifera kept in refrigerator and room temperature for one month showed higher turbidity removal efficiency, compared to those kept for 3 and 5 months, at both containers. The coagulation efficiency of Moringa oleifera was found to be dependent on initial turbidity of water samples. Highest turbidity removals were obtained for water with very high initial turbidity. In summary coagulation efficiency of Moringa oleifera was found independent of storage temperature and container, however coagulation efficiency of Moringa oleifera decreased as storage duration increased. In addition, Moringa oleifera can be used as a potential coagulant especially for very high turbidity water.     

Isolation of a seed coagulant Moringa oleifera lectin

In this work hemagglutinating activity (HA) was investigated in distinct Moringa oleifera tissue extracts. A new lectin from seeds (cMoL) was purified and characterized; hemagglutinating and coagulating activities were evaluated. HA was detected in 0.15 M NaCl extracts from flowers and rachis inflorescence (5%, w/v), seeds, leaves, fundamental tissue of stem and steam bark (10%, w/v). cMoL isolated after saline extraction and guar gel column chromatography was active at pH range 4.0–9.0 agglutinating erythrocytes from rabbit and human blood types. Extracts of tissues and cMoL activities were carbohydrate inhibited; azocasein and asialofetuin abolished cMoL HA. The lectin was thermostable at 100 °C during 7 h. Polyacrylamide gel electrophoresis under reduced conditions revealed a main polypeptide band of 26.5 kDa; native basic cMoL was detected as a unique band. Seed lectin preparations and cMoL showed coagulant activity, similar to aluminium sulphate, the coagulant most widely used in water treatment.     

Soluble Extract from Moringa oleifera Leaves with a New Anticancer Activity

Moringa oleifera has been regarded as a food substance since ancient times and has also been used as a treatment for many diseases. Recently, various therapeutic effects of M. oleifera such as antimicrobial, anticancer, anti-inflammatory, antidiabetic, and antioxidant effects have been investigated; however, most of these studies described only simple biological phenomena and their chemical compositions. Due to the increasing attention on natural products, such as those from plants, and the advantages of oral administration of anticancer drugs, soluble extracts from M. oleifera leaves (MOL) have been prepared and their potential as new anticancer drug candidates has been assessed in this study. Here, the soluble cold Distilled Water extract (4°C; concentration, 300 µg/mL) from MOL greatly induced apoptosis, inhibited tumor cell growth, and lowered the level of internal reactive oxygen species (ROS) in human lung cancer cells as well as other several types of cancer cells, suggesting that the treatment of cancer cells with MOL significantly reduced cancer cell proliferation and invasion. Moreover, over 90% of the genes tested were unexpectedly downregulated more than 2-fold, while just below 1% of the genes were upregulated more than 2-fold in MOL extract-treated cells, when compared with nontreated cells. Since severe dose-dependent rRNA degradation was observed, the abnormal downregulation of numerous genes was considered to be attributable to abnormal RNA formation caused by treatment with MOL extracts. Additionally, the MOL extract showed greater cytotoxicity for tumor cells than for normal cells, strongly suggesting that it could potentially be an ideal anticancer therapeutic candidate specific to cancer cells. These results suggest the potential therapeutic implications of the soluble extract from MOL in the treatment of various types of cancers.     

Enhancing the start-up of a UASB reactor treating domestic wastewater by adding a water extract of Moringa oleifera seeds

Water extract of Moringa oleifera seeds (WEMOS) was used to enhance the start-up of a self-inoculated upflow anaerobic sludge blanket (UASB) reactor treating raw domestic wastewater. Two reactors labelled control (RC) and WEMOS addition (RM) were started without special inoculum. Both reactors were fed continuously for 22 weeks with domestic wastewater containing an average total chemical oxygen demand (COD) of 320 mg O2/l and suspended solid (SS) of 165 mg/l. The reactors operated during the entire experimental period at 29 degrees C and at a hydraulic retention time (HRT) of 4 h. The RM reactor received 2 ml WEMOS per litre of influent. WEMOS solution was prepared on the basis of 2.5% (w/v) ground M. oleifera seeds in water. The results of 22 weeks' operation showed an improvement in the performance of the RM compared to that of the RC. The dosage of WEMOS in the feed (1) shortened the biological start-up period by 20%, (2) increased acidogenic and methanogenic activity by a factor of 2.4 and 2.2 respectively, (3) increased the specific biogas production by a factor of 1.6, (4) favoured fast growth of the sludge bed, and (5) allowed the aggregation of coccoid bacteria and growth of microbial nuclei, which are precursors of anaerobic granulation.     

Moringa oleifera Extract as a Natural Plant Biostimulant

Moringa, belonging to Moringaceae, is known as the “Miracle Tree” that has versatile uses in both animals and plants. The extract from Moringa oliefera serves as a cheap, eco-friendly, novel biostimulator, and bioenhancer that increases sustainable agriculture practices and crop production. Moringa contains several essential components like mineral nutrients, phytohormones (e.g., auxins, gibberellins, and cytokinins), vitamins, flavonols, phenols, sterols, and tannins, as well as several phytochemicals that make it highly beneficial for plants. It induces seed germination, plant growth, photosynthesis, and yields traits at a low cost. It also increases flowering, improves floral traits, fruiting, post-harvesting, and product quality of the fruit, and decreases senescence. Abiotic stresses have a detrimental effect on plant growth and development. The application of Moringa extracts on plants mitigates abiotic stress like salinity, drought, heavy metal, and heat by promoting the activity of antioxidant enzymes and increasing the content of phenols, flavonols, sugars, and osmolyte, which reduces the level of reactive oxygen species, lipid peroxidation, and electrolyte leakage. In particular, Moringa accelerates plant growth, relative water content, water use efficiency, mineral content, gas exchange traits, and yield attributes under stressful environmental conditions. Moringa serves as an essential biopesticide against plant pathogens, and is used in disease management and plant sustenance.

     

Preliminary Crystallographic Analysis of a Cruciferin Protein from Seeds of Moringa oleifera

A 55 kDa cruciferin protein has been purified and characterized from seeds of Moringa oleifera plant. Protein blast of N-terminal amino-acid sequence showed 60 % sequence similarity with cruciferin from Brassica napus. The M. oleifera protein has been crystallized applying the sitting drop method using 5 % polyethylene glycol 8,000, 38.5 % 3-methyl-1,5-pentanediol and 0.1 M sodium cacodylate pH 6.5. The crystals belonged to the P6322 hexagonal space group with cell dimensions, a = b = 98.4, c = 274.3 Å. Initial diffraction data have been collected to a resolution of 6 Å.     

Interactions of surfactants with a water treatment protein from Moringa oleifera seeds in solution studied by zeta-potential and light scattering measurements

Protein extracted from Moringa oleifera (MO) seeds has been advocated as a cheap and environmental friendly alternative to ionic flocculants for water purification. However, the nature and mechanism of its interaction with particles in water, as well as with dissolved surface-active molecules, are not well understood. In this article, we report studies of the protein and its interaction with four surfactants using dynamic light scattering (DLS), zeta-potential and turbidity measurements. Zeta-potential measurements identified points of charge reversal and the turbidity and DLS measurements were used to characterize the microstructure and size of protein-surfactant complexes. From the points of charge reversal, it was estimated that 7 anions are required to neutralize the positive charges of each protein molecule at pH 7. For protein mixtures with sodium dodecyl sulfate and dodecyl di-acid sodium salt, the peak in turbidity corresponds to concentrations with a large change in zeta-potential. No turbidity was observed for protein mixtures with either the nonionic surfactant Triton X-100 or the zwitterionic surfactant N-dodecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate. Changes of pH in the range 4-10 have little effect on the zeta-potential, turbidity, and the hydrodynamic radius reflecting the high isoelectric point of the protein. Addition of small amounts of salt has little effect on the size of protein in solution. These results are discussed in the context of the use of the MO protein in water treatment.     

响应面法优化水酶法提取辣木籽蛋白质的工艺及其功能性质研究

为了提取和利用辣木籽蛋白质,本文通过水酶法优化其提取条件,并探讨其功能性质。以辣木籽为原材料,以蛋白质提取率为指标,首先确定了最佳使用酶为Alcalase碱性蛋白酶,再通过单因素试验考察料液比、时间、温度、酶添加量和pH等因素对蛋白质提取率的影响,在此基础上,利用响应面试验设计优化水酶法提取辣木籽蛋白质的工艺。结果表明,最佳工艺条件为:使用Alcalase碱性蛋白酶,在料液比为1∶10,酶添加量为4.5%,pH为9.0,温度为60℃,时间为4.5 h,此时辣木籽蛋白质的提取率最高为68.23%。在pH为10、温度为55℃时辣木籽蛋白质的氮溶解指数最高;辣木籽蛋白质持水性随着pH的增加而增加,在温度为40℃时持水性最好;在温度为55℃时,辣木籽分离蛋白的吸油效果最明显。     

Moringa oleifera oil: a possible source of biodiesel

Biodiesel is an alternative to petroleum-based conventional diesel fuel and is defined as the mono-alkyl esters of vegetable oils and animal fats. Biodiesel has been prepared from numerous vegetable oils, such as canola (rapeseed), cottonseed, palm, peanut, soybean and sunflower oils as well as a variety of less common oils. In this work, Moringa oleifera oil is evaluated for the first time as potential feedstock for biodiesel. After acid pre-treatment to reduce the acid value of the M. oleifera oil, biodiesel was obtained by a standard transesterification procedure with methanol and an alkali catalyst at 60 degrees C and alcohol/oil ratio of 6:1. M. oleifera oil has a high content of oleic acid (>70%) with saturated fatty acids comprising most of the remaining fatty acid profile. As a result, the methyl esters (biodiesel) obtained from this oil exhibit a high cetane number of approximately 67, one of the highest found for a biodiesel fuel. Other fuel properties of biodiesel derived from M. oleifera such as cloud point, kinematic viscosity and oxidative stability were also determined and are discussed in light of biodiesel standards such as ASTM D6751 and EN 14214. The (1)H NMR spectrum of M. oleifera methyl esters is reported. Overall, M. oleifera oil appears to be an acceptable feedstock for biodiesel.     

Effect of a water extract of Moringa oleifera seeds on the hydrolytic microbial species diversity of a UASB reactor treating domestic wastewater

The effect of a continuous supply of a water extract of Moringa oleifera seeds (WEMOS) on the hydrolytic microbial population of biomass grown in mesophilic upflow anaerobic sludge blanket reactors treating domestic wastewater was investigated. The WEMOS-treated sludge had seemingly a wider diversity, with enterobacter and klebsiella as dominant hydrolytic bacteria, compared with the control sludge. Additional tests indicated that various hydrolytic bacteria could degrade WEMOS. It appeared that a continuous supply of WEMOS to an anaerobic digester, treating domestic wastewater, increased the diversity of hydrolytic bacteria and therefore enhanced the biological start-up of the reactor.     

Potential anti-inflammatory phenolic glycosides from the medicinal plant Moringa oleifera fruits

Bioassay-guided isolation and purification of the ethyl acetate extract of Moringa oleifera fruits yielded three new phenolic glycosides; 4-[(2′-O-acetyl-α-l-rhamnosyloxy) benzyl]isothiocyanate (1), 4-[(3′-O-acetyl-α-l-rhamnosyloxy)benzyl]isothiocyanate (2), and S-methyl-N-{4-[(α-l-rhamnosyloxy)benzyl]}thiocarbamate (3), together with five known phenolic glycosides (4–8). The structures of the new metabolites were determined on the basis of spectroscopic analyses including 1D- and 2D-NMR and mass spectrometry. The anti-inflammatory activity of isolated compounds was investigated with the lipopolysaccharide (LPS)-induced murine macrophage RAW 264.7 cell line. It was found that 4-[(2′-O-acetyl-α-l-rhamnosyloxy)benzyl]isothiocyanate (1) possessed potent NO–inhibitory activity with an IC₅₀ value of 1.67 μM, followed by 2 (IC₅₀ = 2.66 μM), 4 (IC₅₀ = 2.71 μM), and 5 (IC₅₀ = 14.4 μM), respectively. Western blots demonstrated these compounds reduced LPS-mediated iNOS expression. In the concentration range of the IC₅₀ values, no significant cytotoxicity was noted. Structure–activity relationships following NO-release indicated: (1) the isothiocyanate group was essential for activity, (2) acetylation of the isothiocyanate derivatives at C-2′ or at C-3′ of rhamnose led to higher activity, (3) un-acetylated isothiocyanate derivatives displayed eight times less activity than the acetylated derivatives, and (4) acetylation of the thiocarbamate derivatives enhanced activity. These data indicate compounds 1, 2, 4 and 5 are responsible for the reported NO-inhibitory effect of Moringa oleifera fruits, and further studies are warranted.     

Single-step ion exchange purification of the coagulant protein from Moringa oleifera seed

The coagulant protein from Moringa oleifera (MO) seed was purified using a single-step batch ion exchange (IEX) method. Adsorption and elution parameters were optimized. Impact of the purification on the reduction of organic and nutrient release to the water was studied. The matrix was equilibrated using ammonium acetate buffer, and the optimum ionic strength of NaCl for elution was 0.6 M. The time for adsorption equilibrium was between 90 and 120 min. Maximum adsorption capacity of the matrix, estimated with the Langmuir model, was 68 mg protein/g adsorbent. The purified protein does not release organic and nutrient loads to the water, which are the main concerns of the crude extract. This work suggests that a readily scalable single-step IEX purification method can be used to produce the coagulant protein and it can be carried out with locally available facilities. This will promote the use of MO in large water treatment plants and other industries.     

Dimerization of a flocculent protein from Moringa oleifera: experimental evidence and in silico interpretation

Many proteins exist in dimeric and other oligomeric forms to gain stability and functional advantages. In this study, the dimerization property of a coagulant protein (MO_2.1) from Moringa oleifera seeds was addressed through laboratory experiments, protein–protein docking studies and binding free energy calculations. The structure of MO_2.1 was predicted by homology modelling, while binding free energy and residues-distance profile analyses provided insight into the energetics and structural factors for dimer formation. Since the coagulation activities of the monomeric and dimeric forms of MO_2.1 were comparable, it was concluded that oligomerization does not affect the biological activity of the protein.     

Coagulant and antibacterial activities of the water-soluble seed lectin from Moringa oleifera

Aims: The aim of this work was to analyse the coagulant and antibacterial activities of lectin isolated from Moringa oleifera seeds that are used for water treatment. Methods and Results: The water‐soluble M. oleifera lectin (WSMoL) was separated from nonhemagglutinating components (NHC) by chitin chromatography. WSMoL fluorescence spectrum was not altered in the presence of ions that are often present in high concentrations in polluted waters. Seed extract, NHC and WSMoL showed coagulant activity on a turbid water model. Both NHC and WSMoL reduced the growth of Staphylococcus aureus, but only WSMoL caused a reduction in Escherichia coli. WSMoL was also more effective in reducing the growth of ambient lake water bacteria. Conclusions: Data obtained from this study indicate that WSMoL is a potential natural biocoagulant for water, reducing turbidity, suspended solids and bacteria. Significance and Impact of the Study: Moringa oleifera seeds are a material effective in the treatment of water.