Lens Culinaris Seeds Research Articles

Applying a non-GMO breeding approach with an identified natural variation to reduce food allergen Len c3 in Lens culinaris seeds.

Lentils (Lens culinaris) are produced in diverse agroecological regions and are consumed as one of the most important food legumes worldwide. Lentils possess a nutritional profile from a human health perspective that is not only nutrient dense but also offers a better balance between protein and carbohydrates. However, lentil causes food allergy, which has been a significant concern due to increased consumption in parts of the world. Len c3, a non-specific lipid transfer protein (LTP), was identified as one of the allergens in lentil seeds. In this study, we identified an LTP gene Lcu.2RBY.4g013600 that encodes the lentil allergen Len c3. We then focused on gene screening from a collection of natural accessions to search for natural mutations of the Len c3 allergen-encoding gene. A natural lentil line M11 was identified with mutations at LcLTP3b and low accumulation of vicilin through genomic-assisted approaches. Furthermore, we generated a pool of lentil germplasms with LcLTP3b mutation background through crossing the identified lentil plant M11 with two lentil cultivars, CDC Redmoon and CDC Gold. These generated lentil hybrids can be used as a breeding resource targeting at reducing allergen risk in lentil consumption.

Biological treatment of triclosan using a novel strain of Enterobacter cloacae and introducing naphthalene dioxygenase as an effective enzyme

In recent years, triclosan (TCS) has been widely used as an antibacterial agent in personal care products due to the spread of the Coronavirus. TSC is an emerging contaminant, and due to its stability and toxicity, it cannot be completely degraded through traditional wastewater treatment methods. In this study, a novel strain of Enterobacter cloacae was isolated and identified that can grow in high TCS concentrations. Also, we introduced naphthalene dioxygenase as an effective enzyme in TCS biodegradation, and its role during the removal process was investigated along with the laccase enzyme. The change of cell surface hydrophobicity during TCS removal revealed that a glycolipid biosurfactant called rhamnolipid was involved in TCS removal, leading to enhanced biodegradation of TCS. The independent variables, such as initial TCS concentration, pH, removal duration, and temperature, were optimized using the response surface method (RSM). As a result, the maximum TCS removal (97%) was detected at a pH value of 7 and a temperature of 32 °C after 9 days and 12 h of treatment. Gas chromatography-mass spectrometry (GC/MS) analysis showed five intermediate products and a newly proposed pathway for TCS degradation. Finally, the phytotoxicity experiment conducted on Cucumis sativus and Lens culinaris seeds demonstrated an increase in germination power and growth of stems and roots in comparison to untreated water. These results indicate that the final treated water was less toxic.

Thymoquinone Alleviates Cadmium Induced Stress in Germinated Lens culinaris Seeds by Reducing Oxidative Stress and Increasing Antioxidative Activities.

This study investigated the effect of thymoquinone on seeds germination and young seedlings of lentils under cadmium (Cd) stress (300 µM). Three different concentrations (10 µM, 1 µM, and 0.1 µM) of thymoquinone were applied. Our results indicated that thymoquinone has a positive effect on several physiological and biochemical parameters on seeds germination and young seedlings of lentils under Cd stress, which led to enhancing their growth. A significant increase in shoot and root length, fresh and dry weight, and chlorophyll content was observed in the treated plants compared to the control plants. However, the thymoquinone treatment significantly reduced malondialdehyde (MDA) and hydrogen peroxide (H2O2) contents compared to untreated roots and seedlings under Cd-stress. Nevertheless, our results show that the thymoquinone significantly improved the activities of enzymes involved in antioxidant response, including superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), thioredoxin reductase (TrxR), and ascorbate peroxidase (APX). We have also studied the activities of isocitrate dehydrogenase (ICDH) and malate dehydrogenase (MDH); ICDH was increased significantly in roots and seedlings in the presence of different doses of thymoquinone. However, the activity MDH was increased only in roots. Our results suggest that the application of thymoquinone could mitigate cadmium induced oxidative stress.

Antifungal Activity, Structural Stability, and Immunomodulatory Effects on Human Immune Cells of Defensin from the Lentil Lens culinaris.

An increase in the frequency of mycoses and spreading of multidrug-resistant fungal pathogens necessitates the search for new antifungal agents. Earlier, we isolated the novel defensin from lentil Lens culinaris seeds, designated as Lc-def, which inhibited the growth of phytopathogenic fungi. Here, we studied an antifungal activity of Lc-def against human pathogenic Candida species, structural stability of the defensin, and its immunomodulatory effects that may help to prevent fungal infection. We showed that Lc-def caused 50% growth inhibition of clinical isolates of Candida albicans, C. krusei, and C. glabrata at concentrations of 25–50 μM, but was not toxic to different human cells. The lentil defensin was resistant to proteolysis by C. albicans and was not cleaved during simulated gastroduodenal digestion. By using the multiplex xMAP assay, we showed for the first time for plant defensins that Lc-def increased the production of such essential for immunity to candidiasis pro-inflammatory cytokines as IL-12 and IL-17 at the concentration of 2 μM. Thus, we hypothesized that the lentil Lc-def and plant defensins in general may be effective in suppressing of mucocutaneous candidiasis due to their antifungal activity, high structural stability, and ability to activate a protective immune response.

Phytotoxicity of “Tide” Detergent Powder Using Lens culinaris Seeds as a Bioassay

Phytotoxicity of “Tide” Detergent Powder Using Lens culinaris Seeds as a Bioassay

Phytotests for assessing phytotoxicity of “Blue moon” liquid detergent: Lens culinaris seeds

Phytotests for assessing phytotoxicity of “Blue moon” liquid detergent: Lens culinaris seeds

Preliminary pharmacological investigations on Lens culinaris seeds for wound healing potential in rats

Preliminary pharmacological investigations on Lens culinaris seeds for wound healing potential in rats

Ligand Binding Properties of the Lentil Lipid Transfer Protein: Molecular Insight into the Possible Mechanism of Lipid Uptake.

The lentil lipid transfer protein, designated as Lc-LTP2, was isolated from Lens culinaris seeds. The protein belongs to the LTP1 subfamily and consists of 93 amino acid residues. Its spatial structure includes four α-helices (H1-H4) and a long C-terminal tail. Here, we report the ligand binding properties of Lc-LTP2. The fluorescent 2-p-toluidinonaphthalene-6-sulfonate binding assay revealed that the affinity of Lc-LTP2 for saturated and unsaturated fatty acids was enhanced with a decrease in acyl-chain length. Measurements of boundary potential in planar lipid bilayers and calcein dye leakage in vesicular systems revealed preferential interaction of Lc-LTP2 with the negatively charged membranes. Lc-LTP2 more efficiently transferred anionic dimyristoylphosphatidylglycerol (DMPG) than zwitterionic dimyristoylphosphatidylcholine. Nuclear magnetic resonance experiments confirmed the higher affinity of Lc-LTP2 for anionic lipids and those with smaller volumes of hydrophobic chains. The acyl chains of the bound lysopalmitoylphosphatidylglycerol (LPPG), DMPG, or dihexanoylphosphatidylcholine molecules occupied the internal hydrophobic cavity, while their headgroups protruded into the aqueous environment between helices H1 and H3. The spatial structure and backbone dynamics of the Lc-LTP2-LPPG complex were determined. The internal cavity was expanded from ∼600 to ∼1000 Å3 upon the ligand binding. Another entrance into the internal cavity, restricted by the H2-H3 interhelical loop and C-terminal tail, appeared to be responsible for the attachment of Lc-LTP2 to the membrane or micelle surface and probably played an important role in the lipid uptake determining the ligand specificity. Our results confirmed the previous assumption regarding the membrane-mediated antimicrobial action of Lc-LTP2 and afforded molecular insight into its biological role in the plant.

Antihyperlipidemic Activity of Lens culinaris Medikus Seeds in Triton WR- 1339 Induced Hyperlipidemic Rats

Background:Hyperlipidemia involves abnormally elevated lipid levels. Its management using statins, fibrates, bile acid sequestrants and niacin lead to various adverse effects. Lens culinaris (Fabaceae) is a dhal with high nutritional and therapeutic value.Objective: To evaluate antihyperlipidemic activity of Lens culinaris seeds in triton WR-1339 induced hyperlipidemic rats.Materials and methods: Successive extracts, i.e., acetone (LCAE), methanol (LCME) and aqueous (LCWE) were prepared from Lens culinaris Medikus (Fabaceae) seeds. The extracts were evaluated for acute toxicity studies using OECD- 423 guidelines. Their antihyperlipidemic activity was assessed using Triton WR- 1339 induced hyperlipidemia model on Sprague dawley female rats. Animals were divided into 12 groups, i.e., normal, hyperlipidemic (triton), triton+atorvastatin, triton+LCAE, triton+LCME and triton+LCWE groups at a dose of 100, 200, 400 mg/Kg body weight, respectively. Total Cholesterol (TC), Tri Glyceride (TG), High (HDL), Low (LDL) and Very Low Density Lipoprotein (VLDL) levels were assessed after 1, 4, 8 and 24 h of treatment. Total Phenol (TPC), Flavonoid (TFC) and Saponin content were also determined.Results: None of the extracts showed toxicity or mortality to animals. Intragastric administration of extracts at various dose levels to the rats caused a significant decrease in plasma lipid levels. However, LCME (400 mg/Kg) was found to possess more antihyperlipidemic activity as compared to other extracts. It exhibited a decrease (%) of 57.51, 66.93, 66.95, 111.78 in TC, TG, VLDL, LDL levels, and an increase of 59.46% in HDL levels respectively. TPC (608 mg gallic acid equivalent/g of sample) and TFC (128 mg quercetin equivalent/g of sample) were also found highest in LCAE.Conclusion: Results suggest that the extracts of Lens culinaris contain active phytoconstituents which might be responsible for antihyperlipidemic activity of the seeds..

Lens culinaris: A new biocatalyst for reducing carbonyl and nitro groups

A series of aliphatic and aromatic aldehydes and ketones, as well as some nitrocompounds were reduced using whole plant cells from Lens culinaris seeds. In addition, we also investigated the possibility of enzymatic ester hydrolysis to explore the potential of these seeds. The reduced ketones products were obtained in yields of 8 ∼ 82% and enantiomeric excess of 39 ∼ 75%. Aldehydes were more reactive than ketones with high chemical yield (95→99%), whereas the aromatic nitrocompounds showed low (2%) to high (> 99%) conversion depending upon the nature and position of the aromatic ring substituents. Ester hydrolysis by the Lens culinaris was quite effective with the ester p-nitrophenyl acetate (> 99% conversion).

Biochemical and Functional Characterization of an Albumin Protein Belonging to the Hemopexin Superfamily from Lens culinaris Seeds

The present paper reports the purification and biochemical characterization of an albumin identified in mature lentil seeds with high sequence similarity to pea PA2. These proteins are found in many edible seeds and are considered potentially detrimental for human health due to the potential allergenicity and lectin-like activity. Thus, the description of their possible presence in food and the assessment of the molecular properties are relevant. The M(r), pI, and N-terminal sequence of this protein have been determined. The work included the study of (i) the binding properties to hemine to assess the presence of hemopexin structural domains and (ii) the binding properties of the protein to thiamin. In addition, the structural changes induced by heating have been evaluated by means of spectroscopic techniques. Denaturation temperature has also been determined. The present work provides new insights about the structural molecular features and the ligand-binding properties and dynamics of this kind of seed albumin.

A novel defensin from the lentil Lens culinaris seeds

A novel 47-residue plant defensin was purified from germinated seeds of the lentil Lens culinaris by ammonium sulfate precipitation, gel filtration, chromatography, and RP-HPLC. The molecular mass (5440.41 Da) and complete amino acid sequence (KTCENLSDSFKGPCIPDGNCNKHCKEKEHLLSGRCRDDFRCWCTRNC) 1 The protein sequence data reported in this paper will appear in the UniProt Knowledgebase under the Accession No. P85530 and in the GenBank under the accession number EF194158. 1 of defensin, termed Lc-def, were determined. Lc-def has eight cysteines forming four disulfide bonds. The total RNA was isolated from lentil germinated seeds, RT-PCR and subsequent cloning were performed, and cDNA was sequenced. A 74-residue predefensin contains a putative signal peptide (27 amino acid) and a mature protein. Lc-def shows high sequence homology with legumes defensins, exhibits an activity against Aspergillus niger, but does not inhibit proteolytic enzymes.

Influence of germination on lectin in Lens culinaris seeds

The effect of germination conditions on the lectin of Lens culinaris var. Magda 20 seeds was studied. The seeds were germinated at 20 °C under different conditions of watering and light and for different periods of time. The seed lectin was assayed by haemagglutination and quantified by competitive ELISA. Changes in lectin content during germination were also monitored by SDS-PAGE and immunoblotting. Haemagglutinating activity and lectin content in the seeds were not changed during the first three days regardless of the conditions of the germination. However, lectin concentration was significantly higher after six days of germination; relative lectin levels being particularly high when germination was carried out in the light and with daily watering. The results of SDS-PAGE and immunoblotting have also shown that the lectin was not degraded during the first six days of germination however, other storage-proteins were broken down by proteolysis.

X-ray single-crystal structure of 2 (S ),4 (R)-4-hydroxyarginine from Lens culinaris seeds

A new method was developed to isolate, as the hydrochloride salt, 4-hydroxyarginine from Lens culinaris seeds. The structure and absolute configuration of the compound were determined by single-crystal X-ray crystallography and circular dichroism.

Imidazole, a new natural product from the leguminosae

Imidazole was isolated from Lens culinaris seeds as its dansyl derivative and shown to be identical to a synthetic standard by co-chromatography, 1H NMR and EIMS. Chromatographic evidence indicated that it was also present in the seeds of 11 other legumes, 30% of those tested.

α- and β-galactosidase activities in protein bodies and cell walls of lentil seed cotyledons

Cotyledons of mature Lens culinaris seeds contain two forms of both α- and β-galactosidase which can be separated by ion exchange chromatography. These forms are present in cotyledon cell walls and protein bodies except β-galactosidase II,which is undetectable in the cell walls of these organs. All the enzymatic forms were active in an acid pH range but each behaved differently with different substrates, both natural and synthetic, and in the action of different effectors on the activity. α-Galactosidase I and II were able to release free sugars from several putative substrate oligosaccharides and all the forms of α and β-galactosidase were seen to release galactose from lentil storage glycoproteins.

Ultrastructure of Starch Grain Breakdown in Cotyledon Cells of Germinating Lentil Seeds

AbstractFine structural changes associated with the breakdown of starch grains in the storage parenchyma cells of the cotyledons of Lens culinaris seeds during germination are described. In the initial germination phases each starch grain is found inside a double membrane. In more advanced states of germination (8–12 days) the starch grains show erosion from within, leading to the formation of a central cavity surrounded by irregular starch fragments. During this process the starch grain maintains its external shape. Following this it was possible to observe starch fragments dispersed throughout the cytoplasm.

Affinity titration curves Determination of dissociation constants of lectin-sugar complexes and of their pH-dependence by isoelectric focusing electrophoresis

By performing electrophoresis perpendicular to a stationary pH gradient (pH-mobility curves) in polyacrylamide gels containing a specific ligand either covalently fixed or entrapped in the gel matrix, it is possible to measure dissociation constants ( K d) and their pH-dependence in the pH range 3.5–10. The present technique, called ‘affinity titration curves’, is an extension of ‘affinity electrophoresis’. This system has been applied to the study of the interaction between lectins and sugars: lectin from Ricinus communis seeds and α- d-galactose, and lectin from Lens culinaris seeds and α- d-mannose. The pH-dependence of K d values indicated a more rapid decrement of affinity of both lectins for their ligands at acidic pH as compared to alkaline pH. For both lectins, maximum affinity was found in the pH range 7–8. Since the ionic strength of focused carrier ampholytes is 100–200-times lower than in conventional electrophoresis, the K d values found by the present method are generally lower than the same values obtained by affinity electrophoresis.

Some properties of purified phytohemagglutinin from Lens culinaris seeds

Lens culinaris phytohemagglutinin was purified by specific adsorption on Sephadex G-100 and subsequent displacement with d-glucose. The homogeneity of the purified hemagglutinin was ascertained by ultracentrifugal analysis and polyacrylamide gel electrophoresis. The purified hemagglutinin had a s 0 20, w value of 3.4 S. This hemagglutinin was disclosed to be a kind of glycoprotein containing 1.5% glucose and 0.5% glucosamine. The specificity of the purified hemagglutinin was tested by hemagglutination-inhibition assays and found to be essentially the same as that of concanavalin A. In each step of the purification, the mitogenic activity against human peripheral lymphocytes was found to be always confined to the fraction which had the hemagglutinating activity. Morphological studies on hemagglutinin-stimulated cultures indicated that at 72 h about 15% of the cell population had been transformed by the purified hemagglutinin.