Isolation of ß-glucan from Eleusine coracana and its antibiofilm, antidiabetic, antioxidant, and biocompatible activities.

Eleusine coracana (Finger millet) has high nutritional value with numerous health benefits and is of low cost. Isolation of beta-glucan (ßG) from E. coracana (Ec-ßG) has gained increasing research attention. UV-vis spectroscopy used to measure the surface plasmon resonance at 361 nm to confirm the presence of polysaccharides (glucan molecules) in Ec-ßG. X-ray diffraction analysis of Ec-ßG displayed a crystalline nature and confirmed the presence of the ßG molecule. Further, the bioactive compounds of Ec-ßG were screened using gas chromatography-mass spectrometry. The antibacterial activity of Ec-ßG against both Gram-positive (Lysinibacillus fusiformis, Enterococcus faecalis) and Gram-negative (Proteus vulgaris, Shigella sonnei) bacteria were assessed through minimum inhibitory concentrations <70 µg/ml of Ec-ßG. In addition, the antibiofilm activity and bacterial viability of Ec-ßG at 100 µg/ml was confirmed by light and confocal laser scanning microscopy. Furthermore, Ec-ßG inhibits α-amylase and α-glucosidase at an IC50 -value of 1.23 and 1.42 µg/ml, respectively. Superoxide anion scavenging activity at IC50-1.4 µg/ml and DPPH radical scavenging activity at IC50-1.2 µg/ml showed that Ec-ßG had potential antioxidant property. The in vitro hemolysis assay for biocompatibility of Ec-ßG at 200 µg/ml showed 0.06 ± 0.09%. Therefore, Ec-ßG has the potential to act as a suggestive agent for antibacterial, antidiabetic, and antioxidant activity.

Polyhydroxyalkanoate (PHA) biosynthesis from directly valorized ragi husk and sesame oil cake by Bacillus megaterium strain Ti3: Statistical optimization and characterization.

Polyhydroxyalkanoates (PHAs) signify the most promising biological substitute to petrochemical plastics. Renewable and inexpensive agro-industrial by-products can be used as potent fermentation feedstocks for sustainable PHA biosynthesis. This study aimed at using a wild type B. megaterium strain Ti3 innate hydrolytic enzyme/s for eco-friendly valorization of 16 lignocellulosic agrowastes to PHA without pretreatments. Initial hydrolytic screening PHA concentration of (0.04-0.17 g/L), highlighted the strain's metabolic versatility. Pareto ranking of Taguchi orthogonal array (TOA) established ragi husk (RH), sesame oil cake (SOC) and KH2PO4 as the most influential factors (p < 0.05). The optimized and validated Response surface methodology (RSM) model (R2, 0.979; desirability, 1) resulted in 3.8 and 3.6 fold increased PHA production, 4.3 and 3.25 fold increased PHA productivity. A positive correlation (r2, 0.5-0.97) was observed amid the producer innate hydrolytic enzymes (lipase, amylase and cellulase) and PHA production. The PHA was characterized by 1H and 13C NMR, GPC, TGA. The polymer was identified as a scl-mcl copolyester with 92% 3HB (3-hydroxybutyrate) and 8% 3HHp (3-hydroxyheptanoate) monomers by NMR. This the first report on B. megaterium self-enzyme reliant non-food agrowastes bioconversion to PHA with 3HHp (3-hydroxyheptanoate) monomers excluding precursor addition, commercial enzymes, pure carbon and nitrogen sources.

Comparative evaluation of effective microbe- and urea molasses-treated finger millet (Eleusine coracana) straw on nutritive values and growth performance of Washera sheep in northwestern Ethiopia.

An experiment was carried out to comparatively evaluate the effect of effective microbe (EM)- and urea molasses (UM)-treated finger millet straw (EMTFMS and UMTFMS, respectively) on nutrient utilization and growth performance of Washera lambs. Twenty yearling intact male Washera lambs with an initial body weight of 21.13 ± 1.77 kg (mean ± SE) were used for feeding and digestibility trial. These lambs were grouped into five blocks of four animals and randomly assigned to four dietary treatments. The four experimental feeds were untreated finger millet straw (control) + 150 g wheat bran (WB) (T1), untreated finger millet straw + 150 g WB + 150 g Noug seed cake (NSC) (T2), UMTFMS + 150 g WB + 150 g NSC (T3), and EMTFMS + 150 g WB + 150 g NSC (T4). The lambs were de-wormed and vaccinated against ecto- and endo-parasites. Data were analyzed using ANOVA procedure of SAS. Results showed that crude protein (CP) content of finger millet straw (FMS) was improved from 2.13 to 9.7% in UM treatment and 2.13 to 2.39% in EM treatments. Total DM, CP, and OM intake were higher in lambs assigned in UMTFMS (P < 0.01) and EMTFMS (P < 0.01) than the control groups and significantly highest (P < 0.01) in UMTFMS. A non-significant (P > 0.05) difference was observed between treatment technologies in all nutrient apparent digestibility and average daily weight gain. Therefore, it can be concluded that both treatments could serve as an alternative measure to improve nutritive value of finger millet straw, but due to environmental effect, EM treatment could be safe.

Malting Period Effect on the Phenolic Composition and Antioxidant Activity of Finger Millet (Eleusine coracana L. Gaertn) Flour.

The present study examined the influence of malting on the phenolic composition of two cultivars of finger millet using an ultra-performance liquid chromatography mass spectrometer. Total polyphenols and antioxidant activities of the grains were also evaluated using sorghum as an external reference. Catechin, epicatechin, quercetin, taxifolin, and hesperitin were isolated flavonoids, whereas protocatechuic acid was the phenolic acid detected in finger millet malt. Increases in the content of catechin, epicatechin, and protocatechuic acid were observed for 72 h and 96 h for brown finger millet and sorghum malt. Complete loss of taxifolin and hesperitin were observed with the malting period for finger millet cultivars. A similar loss was noted in the proanthocyanidin A1/A2 and catechin content of sorghum with malting time. The grain malt exhibited 2,2-diphenyl-1-picrylhydrazyl,2,2'-azinobis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS) radical scavenging and iron reducing activities. Increased ABTS and iron reducing activity with malting time were observed for the finger millet cultivars. The study demonstrates the presence of hesperitin in finger millet, and also shows that 72 h and 96 h of malting enhanced the catechin, epicatechin, and protocatechuic acid content, in addition to the antioxidant activity of the grain.

Uptake of phenolic compounds from plant foods in human intestinal Caco-2 cells.

In continuation of our studies on the bioaccessibility of phenolic compounds from food grains as influenced by domestic processing, we examined the uptake of phenolics from native/sprouted finger millet (Eleucine coracana) and green gram (Vigna radiata) and native/heat-processed onion (Allium cepa) in human Caco-2 cells. Absorption of pure phenolic compounds, as well as the uptake of phenolic compounds from finger millet, green gram, and onion, was investigated in Caco-2 monolayer model. Transport of individual phenolic compounds from apical compartment to the basolateral compartment across Caco-2 monolayer was also investigated. Sprouting enhanced the uptake of syringic acid from both these grains. Open-pan boiling reduced the uptake of quercetin from the onion. Among pure phenolic compounds, syringic acid was maximally absorbed, while the flavonoid isovitexin was least absorbed. Apparent permeability coefficient P(app) of phenolic compounds from their standard solutions was 2.02 x 10-6cm/s to 8.94 x 10-6cm/s. Sprouting of grains enhanced the uptake of syringic acid by the Caco-2 cells. Open-pan boiling drastically reduced the uptake of quercetin from the onion. The permeability of phenolic acids across Caco-2 monolayer was higher than those of flavonoids.

Evaluation of health potential of nutritionally enriched Kodo millet (Eleusine coracana) grown in Himachal Pradesh, India.

In this study, Kodo millet grains were phytochemically investigated for their nutritional and antioxidant potential for their use as functional foods. Methanolic extracts of grains showed higher phenolic content and antioxidant activity. TLC studies of the extracted polyphenols from kodo millet showed the predominant presence of ferulic acid and cinnamic acid in the millet. Further quantification of these polyphenols was done by using HPLC, analysing ferulic acid and cinnamic acid. Antagonistic spectrum of the polyphenols extracted showed inhibition against four bacterial test indicators viz. Staphylococcus aureus, Leuconostoc mesenteroides, Bacillus cereus and Enterococcus faecalis proving its antimicrobial action. The grains of kodo millet grains taken under study were found to posses' high protein, carbohydrates, minerals, crude fibers, polyphenols and antioxidants thus can be used as a good source of nutrition with additional health benefits.

Identification and molecular characterization of 48 kDa calcium binding protein as calreticulin from finger millet (Eleusine coracana) using peptide mass fingerprinting and transcript profiling.

BACKGROUND: Attempts were made to identify and characterize the calcium binding proteins (CaBPs) in grain filling stages of finger millet using proteomics, bioinformatics and molecular approaches. RESULTS: A distinctly observed blue color band of 48 kDa stained by Stains-all was eluted and analyzed as calreticulin (CRT) using nano liquid chromatography-tandem mass spectrometry (nano LC-MS). Based on the top hits of peptide mass fingerprinting results, conserved primers were designed for isolation of the CRT gene from finger millet using calreticulin sequences of different cereals. The deduced nucleotide sequence analysis of 600 bp amplicon showed up to 91% similarity with CRT gene(s) of rice and other plant species and designated as EcCRT1. Transcript profiling of EcCRT1 showed different levels of relative expression at different stages of developing spikes. The higher expression of EcCRT1 transcripts and protein were observed in later stages of developing spikes which might be due to greater translational synthesis of EcCRT1 protein during seed maturation in finger millet. CONCLUSIONS: Preferentially higher synthesis of this CaBP during later stages of grain filling may be responsible for the sequestration of calcium in endoplasmic reticulum of finger millet grains.

Antioxidant and anti-inflammatory activities of extracts from Cassia alata, Eleusine indica, Eremomastax speciosa, Carica papaya and Polyscias fulva medicinal plants collected in Cameroon.

BACKGROUND: The vast majority of the population around the world has always used medicinal plants as first source of health care to fight infectious and non infectious diseases. Most of these medicinal plants may have scientific evidence to be considered in general practice. OBJECTIVE: The aim of this work was to investigate the antioxidant capacities and anti-inflammatory activities of ethanol extracts of leaves of Cassia alata, Eleusine indica, Carica papaya, Eremomastax speciosa and the stem bark of Polyscias fulva, collected in Cameroon. METHODS: Chemiluminescence was used to analyze the antioxidant activities of plant extracts against hydrogen peroxide or superoxide anion. Comet assays were used to analyze the protection against antioxidant-induced DNA damage induced in white blood cells after treating with hydrogen peroxide. Flow cytometry was used to measure γδ T cells proliferation and anti-inflammatory activity of γδ T cells and of immature dendritic cells (imDC) in the presence of different concentrations of plant extracts. RESULTS: Ethanol extracts showed strong antioxidant properties against both hydrogen peroxide and superoxide anion. Cassia alata showed the highest antioxidant activity. The effect of plant extracts on γδ T cells and imDC was evidenced by the dose dependent reduction in TNF-α production in the presence of Cassia alata, Carica papaya, Eremomastax speciosa Eleusine indica, and Polyscias fulva. γδ T cells proliferation was affected to the greatest extent by Polyscias fulva. CONCLUSION: These results clearly show the antioxidant capacity and anti-inflammatory activities of plant extracts collected in Cameroon. These properties of leaves and stem bark extracts may contribute to the value for these plants in traditional medicine and in general medical practice.

Bioaccessible nutrients and bioactive components from fortified products prepared using finger millet (Eleusine coracana).

BACKGROUND: Finger millet (Eleusine coracana), a staple food in semi-arid parts of the world, is a rich source of nutrients and bioactive components comparable to rice and wheat but with higher fibre content. Unprocessed and processed finger millet (whole flour (WFM), sieved flour (SFM), wafers and vermicelli with altered matrices (added Fe or Zn or reduced fibre)) were analysed for chemical composition, bioaccessible Fe, Zn and Ca, in vitro digestible starch (IVSD) and protein (IVPD) and bioactive components (polyphenols and flavonoids). RESULTS: WFM and SFM flours differed significantly in their composition. Sieving decreased the content of both nutrients and antinutrients in WFM but increased their digestibility/bioaccessibility. WFM products with Zn and Fe showed highest IVPD, whereas SFM products with Fe showed highest IVSD. Products with externally added Fe and Zn showed maximum bioaccessibility of Fe and Zn respectively. WFM had the highest levels of total polyphenols and flavonoids, 4.18 and 15.85 g kg⁻¹ respectively; however, bioaccessibility was highest in SFM vermicelli. CONCLUSION: The availability of nutrients and bioactive components was influenced by both processing methods and compositional alterations of the food matrix in finger millet products, and bioaccessibility of all constituents was higher in vermicelli (wet matrix) than in wafers (dry matrix).

Purification and characterization of fructan and fructansucrase from Lactobacillus fermentum AKJ15 isolated from Kodo ko jaanr, a fermented beverage from north-eastern Himalayas.

Fructansucrase and fructan produced from Lactobacillus fermentum AKJ15 were isolated from seeds of Kodo ko jaanr, a fermented mild-alcoholic beverage prepared in North East India. The strain was identified by 16S rRNA gene sequence analysis and biochemical characterization. The strain displayed maximum fructansucrase activity of 4.3 U/ml (1.02 U/mg) at 28°C at 180 rpm. The enzyme purified by polyethylene glycol-400 gave specific activity of 5 U/mg and showed 90 kDa band on non-denaturing Sodium Dodecyl Sulphate-Poly Acrylamide Gel Electrophoresis (SDS-PAGE). The purified enzyme confirmed the presence of fructan by periodic acid Schiff's staining which showed magenta colour bands with both sucrose and raffinose. The strain produced 10.2 mg/ml fructan in broth under optimized culture conditions. The purified fructansucrase displayed V(max) of 5.42 U/mg and K(m) of 16.65 mM. The enzyme showed maximum activity at 30°C and at pH 5. The structure of fructan was analysed by (1)H and (13)C NMR spectra confirming ß-(2-1) and ß-(2-6) linkages.

Eleusine indica L. possesses antioxidant activity and precludes carbon tetrachloride (CCl4)-mediated oxidative hepatic damage in rats.

OBJECTIVES: The purpose of this study was to evaluate the ability of aqueous extract of Eleusine indica to protect against carbon tetrachloride (CCl4)-induced hepatic injury in rats. METHODS: The antioxidant activity of E. indica was evaluated using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay. The total phenolic content of E. indica was also determined. Biochemical parameters [e.g. alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA), glutathione (GSH), catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase and quinone reductase] were used to evaluate hepatic damage in animals pretreated with E. indica and intoxicated with CCl4. CCl4-mediated hepatic damage was also evaluated by histopathologically. RESULTS: E. indica extract was able to reduce the stable DPPH level in a dose-dependent manner. The half maximal inhibitory concentration (IC50) value was 2350 µg/ml. Total phenolic content was found to be 14.9 ± 0.002 mg/g total phenolic expressed as gallic acid equivalent per gram of extract. Groups pretreated with E. indica showed significantly increased activity of antioxidant enzymes compared to the CCl4-intoxicated group (p < 0.05). The increased levels of serum ALT and AST were significantly prevented by E. indica pretreatment (p < 0.05). The extent of MDA formation due to lipid peroxidation was significantly reduced (p < 0.05), and reduced GSH was significantly increased in a dose-dependently manner (p < 0.05) in the E. indica-pretreated groups as compared to the CCl4-intoxicated group. The protective effect of E. indica was further evident through decreased histopathological alterations in the liver. CONCLUSION: The results of our study indicate that the hepatoprotective effects of E. indica might be ascribable to its antioxidant and free radical scavenging property.

Finger millet (Eleucine coracana) flour as a vehicle for fortification with zinc.

Millets, being less expensive compared to cereals and the staple for the poorer sections of population, could be the choice for fortification with micronutrients such as zinc. In view of this, finger millet, widely grown and commonly consumed in southern India, was explored as a vehicle for fortification with zinc in this investigation. Finger millet flour fortified with either zinc oxide or zinc stearate so as to provide 50mg zinc per kg flour, was specifically examined for the bioaccessibility of the fortified mineral, as measured by in vitro simulated gastrointestinal digestion procedure and storage stability. Addition of the zinc salts increased the bioaccessible zinc content by 1.5-3 times that of the unfortified flour. Inclusion of EDTA along with the fortified salt significantly enhanced the bioaccessibility of zinc from the fortified flours, the increase being three-fold. Inclusion of citric acid along with the zinc salt and EDTA during fortification did not have any additional beneficial effect on zinc bioaccessiblity. Moisture and free fatty acid contents of the stored fortified flours indicated the keeping quality of the same, up to 60 days. Both zinc oxide and zinc stearate were equally effective as fortificants, when used in combination with EDTA as a co-fortificant. The preparation of either roti or dumpling from the fortified flours stored up to 60 days did not result in any significant compromise in the bioaccessible zinc content. Thus, the present study has revealed that finger millet flour can effectively be used as a vehicle for zinc fortification to derive additional amounts of bioaccessible zinc, with reasonably good storage stability, to combat zinc deficiency.

Storage behavior of immune-enhancing enteral formulation from natural sources.

Nutrition support has become an important therapeutic intervention for improving outcomes in hospitalized patients. The present study was carried out with the objective of studying appropriate packaging and storage studies of enteral formulation from natural sources comprising finger millet (Eleusine coracana), soyprotein isolate, tomato powder, carrot powder, anne greens (Celosia argentea), groundnut oil and fish oil. Two type of packages--namely, polypropylene pouches and metallized polyester/polyethylene pouches--were used. The storage conditions were 27°C and 42°C at 65% relative humidity. The chemical, microbiological and sensory qualities of the formulation were assessed at regular intervals. The moisture sorption studies revealed that the formulation had an initial moisture content of 3.59%, which equilibrated to 13%. The product was acceptable at 64% relative humidity, after which it tend to become soggy. Analysis of peroxide value and free fatty acid content of the equilibrated samples revealed that moisture content of 4-5% was ideal for the storage of the formulation. An increase in the peroxide value and free fatty acid content was observed during 90-day storage period but the formulation was organoleptically acceptable. The microbial analysis of the formulation revealed an initial total bacterial count of 1.5 × 10² colony-forming units and increased to 7.9 × 10² colony-forming units (polypropylene) and 5.0 × 10² colony-forming units (metallized polyester/polyethylene). No fungal and coliform was detected at 90 days of storage. Thus it was concluded that the formulation can be stored for a period of 3 months at 27°C and 65% relative humidity. Such formulations can be primarily a supportive strategy to an active therapeutic intervention.