Characterisation of lipoxygenase from pea seeds (Pisum sativum var. Telephone L.)

Lipoxygenase (LOX) from pea seeds (Pisum sativum var. Telephone L.) was extracted and studied of biochemical properties. The molecular mass of purified lipoxygenase was 93 kDa. The effects of substrate specificity, pH, and sensibility to various inhibitors: caffeic acid, ferulic acid, benzoic acid, catechin, quercetin and kaempferol of LOX were investigated. Lipoxygenase showed the highest activity toward linoleic acid and the lowest toward oleic acid as substrates. Kinetic studies indicated that V_max of the LOX activity was 151.5 U/min and corresponding K_m value of 0.44 × 10⁻³ M. Optimum pH of lipoxygenase was reported at 5.5. Caffeic acid was the most effective inhibitor and kaempferol was the least effective.     

Antifungal and antibacterial activities of lectin from the seeds of Archidendron jiringa Nielsen

A Archidendron jiringa Nielsen lectin was purified by aqueous extraction, 90% ammonium sulphate precipitation and concanavalinA-Sepharose 4B affinity chromatography. Its specific activity was of 88.3 × 10² hemagglutination unit/mg protein for a yield of 51.6% total protein. The molecular weight is of 35.7 kDa. It has hemagglutinating activity against human blood group, rabbit, mouse, rat, guinea pig, geese and sheep erythrocytes. The hemagglutination activity of lectin was relatively insensitive to acidic pH above 2, had an optimal activity at pH 8, and stable below 45 °C for 30 min. The activity was stimulated by Ca²⁺, Mg²⁺ and Mn²⁺. The internal sequence indicated similarity with legume lectin family. Moreover, even at low concentrations antifungal activity was observed against Exserohilum turcicum, Fusarium oxysporum and Colletotrichum cassiicola. The minimal inhibitory concentrations were 0.227, 0.0567 and 0.0567 mg/ml for Bacillus subtilis, Staphylococcus aureus, and Candida albicans, respectively.     

Chemical composition and antimicrobial activity of the essential oil of Coriandrum sativum

The essential oil from leaves of Coriandrum sativum L. (Apiaceae), obtained by hydro-distillation was analysed by gas chromatography–mass spectrometry (GC–MS) and also evaluated for in vitro antimicrobial activity. Out of 27 peaks, 24 components, which constitute 92.7%, were identified in the oil. The oil was dominated by aldehydes and alcohols which accounted for 56.1% and 46.3% of the oil, respectively. The major constituents were 2E-decenal (15.9%), decanal (14.3%), 2E-decen-1-ol (14.2%) and n-decanol (13.6%). Other constituents present in fairly good amounts are 2E-tridecen-1-al (6.75%), 2E-dodecenal (6.23%), dodecanal (4.36%), undecanol (3.37%), and undecanal (3.23%). The oil was screened for antimicrobial activity against both Gram positive (Staphylococcus aureus, Bacillus spp.) and Gram negative (Escherichia coli, Salmonella typhi, Klebsiella pneumonia, Proteus mirabilis, Pseudomonas aeruginosae) bacteria and a pathogenic fungus, Candida albicans. The oil showed pronounced antibacterial and antifungal activity against all of the microbes tested, except for P. aeruginosae, which showed resistance.     

Effect of Pisum sativum fractions on the mortality and progeny production of nine stored-grain beetles

Yellow field pea (Pisum sativum L.) fractions that were mainly protein (50%), fibre (90%) or starch (85%) were obtained from a commercial pea mill and mixed with wheat kernels or wheat flour. Based on the mortality and the number of offspring produced, protein-rich pea flour was more toxic than fibre, which was more toxic than starch. For the protein-rich pea flour mixed with wheat kernels, the most sensitive insects were Sitophilus oryzae (L.), Sitophilus zeamais Motschulsky and Sitophilus granarius (L.), followed by Cryptolestes ferrugineus (Stephens) which was more sensitive than Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.). For the protein-rich pea flour mixed with wheat flour, Cryptolestes pusillus (Schönherr) was most sensitive, followed by C. turcicus (Grouvelle) and T. confusum (Jacquelin du Val), with T. castaneum being the most resistant. Although protein-rich pea flour did not kill adults to a great extent when mixed with flour, it reduced offspring production significantly. Again C. pusillus was the most sensitive, followed by T. confusum, with T. castaneum offspring being the most resistant. The insecticidal activity of pea fractions decreased after treated wheat kernels were held at 30 °C, 70% r.h. for 8 months. The potential of using pea fractions to control stored-product insects is discussed.     

Chemical composition and antifungal activity of essential oils from three Himalayan Erigeron species

Three Himalayan Erigeron (Asteraceae) species viz Erigeron mucronatus, Erigeron annuus and Erigeron karwinskianus growing in sub-alpine region revealed occurrence of isomeric polyacetylenic constituents viz., matricaria and lachnophyllum esters which accounted for 83.3%, 69.3% and 30.1% of the essential oils from these species, respectively, in addition to mono- and sesquiterpenoids as minor constituents. The antifungal activity tested by poisoned food (PF) techniques against Fusarium oxysporum, Helminthosporium maydis, Rhizoctonia solani, Alternaria solani and Sclerotinia sclerotiorum demonstrated significant inhibition of the mycelial growth of all strains (p < 0.05). The oils (500 μg/mL) showed significant antifungal effect against tested fungi in the growth inhibition range of 37.6–85.5% with respective IC₅₀ values ranging from 88.8 to 660.0 μg/mL as compared to standard fungicides (100% inhibition) with IC₅₀ value in the range of 32.2–129.4 μg/mL. Significant inhibition of spore germination was noticed for F. oxysporum, Curvularia lunata and Albugo candida which were highly susceptible to E. annuus oil with their IC₅₀ values 120.7, 253.5 and 300.4 μg/mL, respectively. Thus, the results obtained in this study demonstrate the potential of essential oils from Himalayan Erigeron species as non-toxic, eco-friendly and biodegradable natural fungicides.     

Antimicrobial activity of essential oil extracts of various onions (Allium cepa) and garlic (Allium sativum)

Antimicrobial activity of different concentrations (50, 100, 200, 300 and 500 ml/l) of essential oil extracts of three type of onions (green, yellow and red) and garlic against two bacteria, Staphylococcus aureus, Salmomella Enteritidis, and three fungi, Aspergillus niger, Penicillium cyclopium and Fusarium oxysporum, was investigated. The essential oil (EO) extracts of these Allium plants (garlic and onions) exhibited marked antibacterial activity, with garlic showing the highest inhibition and green onion the lowest. Comparatively, 50 and 100 ml/l concentrations of onions extracts were less inhibitory than 200, 300 and 500 ml/l concentrations. However, with garlic extract, high inhibitory activity was observed for all tested concentrations. S. aureus showed less sensitivity towards EO extracts inhibition, however S. Enteritidis was strongly inhibited by red onion and garlic extracts. The fungus F. oxysporum showed the lowest sensitivity towards EO extracts, whereas A. niger and P. cyclopium were significantly inhibited particularly at low concentrations. Conclusively, where seasoning is desired, essential oil extracts of onions and garlic can be used as natural antimicrobial additives for incorporating in various food products.     

Antimicrobial potential of Coriandrum sativum L. against different Candida species in vitro

The aim of this study was to evaluate the chemical and antifungal activity of the essential oil from Coriandrum sativum L. (Apiaceae) against different Candida species. The essential oil (EO) was obtained by hydrodistillation and submitted to dry-column chromatography, resulting in six fractions, which were then submitted to TLC and GC–MS analysis. The main compounds identified were alcohols: 1-decanol (24.20%); 2E-decenol (18.00%); 2Z-dodecenol (17.60%); and aldehydes (89%). Antibacterial activity of the EO and its fractions was tested against five species of Candida albicans. The EO showed antimicrobial activity against all the species of Candida tested, except for Candida tropicalis CBS 94. Fractions 4 and 6 had a greater antibiotic spectrum, probably due to the presence of such alcohols as 3-hexenol, 1-decanol, 2E-decenol and 2Z-dodecenol. In conclusion, the EO and its fractions could be used as potential antimicrobial agents to treat or prevent Candida yeast infections.     

Purification and partial characterization of polyphenol oxidase from the flower buds of Lonicera japonica Thunb.

The purification and partial enzymology characteristics of polyphenol oxidase from Lonicera japonica (LjPPO) were studied in this paper. The crude enzyme solution was purified in turn by ammonium sulfate, dialysis, and DEAE-cellulose ion-exchange chromatography after preliminary treatments. Purification resulted in 31-fold enrichment and its molecular weight was estimated to be ∼49 kDa exhibited on sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). The pH for optimal conditions of LjPPO was 7.5, and the temperature was 25 °C, in addition, the inhibitive effects of inhibitors were enhanced positively with increasing of the concentration. Moreover, crude enzyme solution showed diphenolase activity toward catechol, l-dopa and chlorogenic acid rather than monophenolase and triphenolase activity, and the best substrate was catechol because of the highest V_max/K_m value. However, the oxidation of diphenol related to browning significantly, so the data obtained in this research provided theoretical basis for the prevention of enzymatic browning of L. japonica during processing.     

Isolation, purification and characterization of a new gum from Acanthophyllum bracteatum roots

A new gum was isolated from the roots of Acanthophyllum bracteatum (ABG) by warm-water extraction. Purification was carried out by barium complexing to give a yield of 12.4% of pure air-dried or 5.8% of freeze-dried gum. The ABG contained 13.2% moisture, 84.3% carbohydrate, 0.9% protein and 1.5% ash. Its mineral content was comparable to commercial hydrocolloids. Monosaccharide analysis by HPLC showed the presence of galactose, glucose, arabinose, rhamnose and uronic acids in the ratio 16.0:7.2:3.0:1.0:3.1 respectively. The viscosity and pH value of 1% ABG solution at 25 °C were 51.5 mPa s and 6.85 respectively. ABG solutions (5-30 wt%) showed shear-thinning flow behavior at shear rates < 10 s⁻¹. The viscosity decreased as temperature increased, and was highest at the neutral state. ABG had low surface and emulsification properties but moderate foaming capacity and relatively high foaming stability, which suggests that ABG could potentially be used in food systems to improve foaming properties.     

Preparation,preliminary characterization and immunostimulatory activity of polysaccharide fractions from the peduncles of Hovenia dulcis

The peduncles of Hovenia dulcis, containing abundant nutrients and having a taste like a combination of raisin, clove, cinnamon and sugar, have been consumed as fruits and used as traditional herbal medicine for a long time in China. Up to date, little information is available about the polysaccharides from peduncles of H. dulcis (HDPS) and their potential bioactivity. In this study, three purified fractions were prepared by sequential purification of crude HDPS through ion-exchange chromatography and gel-filtration chromatography. The three fractions of HDPS-1, HDPS-2 and HDPS-3 were found to be homogeneous heteropolysaccharides mainly composed of rhamnose, arabinose, galactose and galacturonic acid with an average molecular weight of 235, 70 and 53 kDa, respectively. HDPS-3 was quite different from HDPS-1 and HDPS-2, as it contained much higher content of galacturonic acid (40.5%). In vitro immunostimulatory activity evaluation revealed that all the three fractions could significantly stimulate the proliferation of splenocytes and enhance phagocytosis, nitric oxide production and acid phosphatase activity of peritoneal macrophages, which suggested that HDPS had a potent immunostimulatory activity and could be explored as a potential natural immunomodulatory agent.     

Phenolic profiles of in vivo and in vitro grown Coriandrum sativum L.

Coriandrum sativum L. is a source of a variety of polyphenols and other phytochemicals, related to its high antioxidant activity and to its use for indigestion, rheumatism, and prevention of lipid peroxidation damage. Plant cell cultures are a means to study or to produce some active metabolites, such as polyphenols. This technique was applied to the investigation of coriander, and a detailed analysis of individual polyphenols in vivo and in vitro grown samples was performed. The in vivo vegetative parts showed quercetin derivatives as the main flavonoids and quercetin-3-O-rutinoside (3296 mg/kg dw) was the main polyphenol found in this part of coriander. The fruits revealed only phenolic acids and derivatives, caffeoyl N-tryptophan hexoside (45.33 mg/kg dw) being the most abundant phenolic derivative. In vitro samples also gave a high diversity of polyphenols, being C-glycosylated apigenin (2983 mg/kg dw) the main compound. Anthocyanins were only found in clone A, which was certainly related to its purple pigmentation, and peonidin-3-O-feruloylglucoside-5-O-glucoside was the major anthocyanin found (1.70 μg/kg dw). In vitro culture can be used to explore new industrial, pharmaceutical, and medicinal potentialities, such as the production of secondary metabolites like flavonoids.     

Purification,preliminary structural characterization and in vitro antioxidant activity of polysaccharides from Acanthus ilicifolius

Crude Acanthus ilicifolius (A. ilicifolius) polysaccharides (CAIP) were obtained by hot water extraction and deproteinated. Two major polysaccharide fractions, the neutral A. ilicifolius polysaccharide (NAIP) and the acid A. ilicifolius polysaccharide B (AAIP-B), were isolated from CAIP by chromatography using DEAE-Sepharose Fast Flow and Sepharose CL-6B. The molecular weights (M_w) of NAIP and AAIP-B, determined by high performance gel-filtration chromatography (HPGFC), were 11,775 and 23,161 Da, respectively. AAIP-B contained 51.23% uronic acid, characteristic of a pectin-type hetero-polysaccharide. Analysis of the neutral monosaccharide composition indicated that NAIP contained high proportions of arabinose, galactose and glucose. However, AAIP-B was mainly composed of rhamnose, arabinose and galactose. Their structure properties were confirmed by FT-IR.     

Extraction optimization of a hydrocolloid extract from cress seed (Lepidium sativum) using response surface methodology

Extraction conditions for maximum values of yield, viscosity and minimum protein content of hydrocolloid extract from Lepidium sativum seed were investigated using response surface methodology. A Central Composite Face Design (CCFD) with four independent variables: temperature (25–85 °C); pH (3–10); extraction time (10–25 min) and water to seed ratio (10:1–80:1) was used to study the response variables (yield, viscosity and protein content). Data analysis showed that all the variables significantly (p < 0.05) affected the extraction yield and viscosity, whereas the effect of water to seed ratio on protein content was not significant (p > 0.05). Applying a desirability function method the optimum parameters were: extraction temperature 35 °C, pH 10, water to seed ratio of 30:1 and an extraction time of 15 min. At this optimum point, apparent viscosity, yield and protein content were 0.2 Pa s, 6.46% and 0.57%, respectively. The experimental values were very close to the predicted values and were not statistically different (p > 0.05).     

Isolation and characterization of galactomannan from Dimorphandra gardneriana Tul. seeds as a potential guar gum substitute

A galactomannan was obtained from mature seeds of Dimorphandra gardneriana Tul., the plant from which rutin is extracted. The galactomannan extraction was based on manual separation of the endosperm, water dissolution, centrifugation and precipitation with ethanol. The galactomannan yield obtained (31%) was similar to values reported for other Brazilian seeds and to that of guar gum. The polysaccharide from D. gardneriana seeds (GalDG) was characterized by gas–liquid chromatography (GLC), gel permeation chromatography (GPC), rheology and also by ¹³C and ¹H nuclear magnetic resonance (NMR). The monosaccharide composition in weight % was mannose 64.2, galactose 34.7 and glucose 1.1. Small amounts of protein and uronic acid were found, values being 1.75 and 2.8% (w/w), respectively. The mannose/galactose ratio of GalDG (1.84) is similar to values reported for galactomannans extracted from other Brazilian seeds, and is the M/G value closest to that of guar gum (1.6–1.8). The intrinsic viscosity of galactomannan from D. gardneriana (8.7 dL/g), in water at 25 °C, is lower than the [η] value of guar gum, but the absolute viscosity of the GalDG in aqueous solution at concentrations of 0.1 and 1% (w/v) is higher. The aqueous solution at 1% (w/v) behaves as a pseudoplastic fluid, but a Newtonian behavior was noted for the solution at 0.1%. The high average molar masses, M_w of 3.9 × 10⁷ g/mol and M_n of 1.9 × 10⁷ g/mol, determined by GPC are probably due to molecular aggregation. ¹³C and ¹H NMR spectra (DEPT 135 and HSQC) of GalDG solutions in D₂O were recorded. The patterns of mannose substitution in GalDG and guar gum are similar.     

Antioxidant Activity of Pea (Pisum sativum L.) Seed Coat Acetone Extract

The contents of phenolic compounds in seed coat of pea and their antioxidative properties were examined. The pea seed coat was extracted with acetone-water (7:3 v/v) mixture and the extract was separated into five (?V) fractions using a Sephadex LH-20 column chromatography. Antioxidative activity of extract and fractions was measured by the oxidation of phosphatidylcholine to hydroxyperoxidephosphatidylcholine in liposome model and by scavenging effects of superoxide radical anion in xanthine-xanthine oxidase system. Phenolic compounds of extract and fractions were determined by spectrophotometric methods and characterized by HPLC analysis. Strong antioxidative properties were noted for extract and its five fractions measured by liposome method. The extract and fractions I, IV and V also showed scavenging effects of superoxide radical anion. A statistically significant correlation (P≤0.05) was found between the inhibition of PC oxidation in the system tested and contents of either total phenols or tannins. However no statistically significant correlation was found between O^•−₂ scavenging effect and contents of either total phenols or tannins. The HPLC analysis of phenolic compounds of extract and active fractions showed the presence of some phenolic acids (benzoic and cinnamic acids, and cinnamic acid derivatives), flavone and flavonol glycoside.     

Spirostane,furostane and cholestane saponins from Persian leek with antifungal activity

A phytochemical investigation of the seeds of Persian leek afforded the isolation of two new spirostane glycosides, persicosides A (1) and B (2), four new furostane glycosides, isolated as a couple of inseparable mixture, persicosides C1/C2 (3a/3b) and D1/D2 (4a/4b), one cholestane glycoside, persicoside E (5), together with the furostane glycosides ceposides A1/A2 and C1/C2 (6a/6b and 7a/7b), tropeosides A1/A2 and B1/B2 (8a/8b and 9a/9b), and ascalonicoside A1/A2 (10a/10b), already described in white onion, red Tropea onion, and shallot, respectively. Structure elucidation of the compounds was carried out by comprehensive spectroscopic analyses, including 2D NMR spectroscopy and MS spectrometry, and by chemical evidences. The chemical structure of new compounds were identified as (25S)-spirostan-2α,3β,6β-triol 3-O-[β-d-glucopyranosyl-(1 → 3)] [β-d-xylopyranosyl-(1 → 2)]-β-d-glucopyranosyl-(1 → 4)-β-d-galactopyranoside (1), (25S)-spirostan-2α,3β,6β-triol 3-O-[β-d-xylopyranosyl-(1 → 3)] [α-l-rhamnopyranosyl-(1 → 2)]-β-d-glucopyranosyl-(1 → 4)-O-β-d-galactopyranoside (2), furosta-1β,3β,22ξ,26-tetraol 5-en 1-O-β-d-glucopyranosyl (1 → 3)-β-d-glucopyranosyl (1 → 2)-β-d-galactopyranosyl 26-O-α-l-rhamnopyranosyl (1 → 2)-β-d-galactopyranoside (3a,3b), furosta-2α,3β,22ξ,26-tetraol 3-O-β-d-glucopyranosyl (1 → 3)-β-d-glucopyranosyl (1 → 2)-β-d-galactopyranosyl 26-O-β-d-glucopyranoside (4a,4b), (22S)-cholesta-1β,3β,16β,22β-tetraol 5-en 1-O-α-l-rhamnopyranosyl 16-O-α-l-rhamnopyranosyl (1 → 2)-β-d-galactopyranoside (5).     

Salinity impact on fruit yield,essential oil composition and antioxidant activities of Coriandrum sativum fruit extracts

This study is designed to examine the fruit essential oil composition, the total phenolic amounts and the antioxidant activities in methanolic extracts of Coriandrum sativum under saline conditions. Increasing NaCl levels to 75 mM reduced significantly the fruit yield by 36%. The essential oil yield was 0.30%, based on the dry weight; it increased by 77% and 84% at 50 and 75 mM NaCl, respectively, in comparison to the control. The major constituents were linalool and camphor, whose amounts increased with increasing NaCl concentrations. Antioxidant activities of the methanol extracts were determined by three different test systems, namely DPPH, β-carotene/linoleic acid and reducing power assays. In these three test systems, the highest activity was exhibited in control plants and was reduced significantly with increasing NaCl levels. In control plants, the total phenolic amount was 1.04 mg GAE/g DW which decreased by 43% and 66% at 50 and 75 mM NaCl, respectively.     

In vitro and in vivo antifungal activities of the essential oils of various plants against tomato grey mould disease agent Botrytis cinerea

The aim of this study was to find an alternative to synthetic fungicides currently used in the control of devastating fungal pathogen Botrytis cinerea, the causal agent of grey mould disease of tomato. Antifungal activities of essential oils obtained from aerial parts of aromatic plants, which belong to the Lamiacea family such as origanum (Origanum syriacum L. var. bevanii), lavender (Lavandula stoechas L. var. stoechas) and rosemary (Rosmarinus officinalis L.), were investigated against B. cinerea. Contact and volatile phase effects of different concentrations of the essential oils were found to inhibit the growth of B. cinerea in a dose-dependent manner. Volatile phase effects of essential oils were consistently found to be more effective on fungal growth than contact phase effect. A volatile vapour of origanum oil at 0.2 μg/ml air was found to completely inhibit the growth of B. cinerea. Complete growth inhibition of pathogen by essential oil of lavender and rosemary was, however, observed at 1.6 μg/ml air concentrations. For the determination of the contact phase effects of the tested essential oils, origanum oil at 12.8 μg/ml was found to inhibit the growth of B. cinerea completely. Essential oils of rosemary and lavender were inhibitory at relatively higher concentrations (25.6 μg/ml). Spore germination and germ tube elongation were also inhibited by the essential oils tested. Light and scanning electron microscopic (SEM) observations revealed that the essential oils cause considerable morphological degenerations of the fungal hyphae such as cytoplasmic coagulation, vacuolations, hyphal shrivelling and protoplast leakage and loss of conidiation. In vivo assays with the origanum essential oil, being the most efficient essential oil, under greenhouse conditions using susceptible tomato plants resulted in good protection against grey mould severity especially as a curative treatment. This study has demonstrated that the essential oils are potential and promising antifungal agents which could be used as biofungicide in the protection of tomato against B. cinerea.