Structural characterisation of polysaccharides from Tricholoma matsutake and their antioxidant and antitumour activities

In this study, polysaccharides from Tricholoma matsutake (TM-P) were purified using a DEAE-Sepharose fast flow column and three polysaccharide fractions (TM-P1, TM-P2 and TM-P3) were obtained. The chemical composition and structural characteristics of TM-P2 were quite different from those of TM-P1 and TM-P3. TM-P2 consisted of glucose, galactose and mannose with a molar ratio of 5.9:1.1:1.0. The glycosidic linkages were mainly composed of 1,6- and 1-linked glucose. Furthermore, TM-P2 showed the strongest in vitro antioxidant and antitumour activities. The oxygen radical absorbance capacity (ORAC) of TM-P2 was 2100.44 μmol Trolox/g. The antiproliferative activities of TM-P2 (4.0 mg/ml) on the growth of HepG2 and A549 cells were 67.98% and 59.04%, respectively.     

Comparison of umami-taste active components in the pileus and stipe of pine-mushrooms (Tricholoma matsutake Sing.) of different grades

Umami-taste active components in pine-mushrooms (Tricholoma matsutake Sing.) were compared according to different grades (first, second, third, and fourth) and parts (pileus and stipe). The contents of umami-taste active free amino acids, such as aspartic acid and glutamic acid, decreased in the order of second grade > third grade > fourth grade > first grade. Also, the contents of umami-taste active 5′-nucleotides were highest and lowest in pine-mushrooms of second and first grades, respectively. All the contents of umami-taste active amino acids and 5′-nucleotides were higher in the pileus than in the stipe, irrespective of pine-mushroom grades. Equivalent umami concentration values of pine-mushrooms ranged from 13.26 (in the stipe of first grade) to 204.26 g (in the pileus of second grade) per 100 g. All the quantitative data obtained in this study are highly consistent with our previously data on umami sensory intensities of pine-mushrooms.     

Process development for mycelial growth and polysaccharide production in Tricholoma matsutake liquid culture

In this study, the effects of agitation and aeration on mycelial growth and exo-polysaccharide production were examined in batch cultures of Tricholoma matsutake. Agitation was varied from 100 to 300 rpm and aeration was varied from 0.5 to 1.5 vvm. Mycelial growth was 21.87 g/l at 150 rpm, and exo-polysaccharide production was 8.79 g/l at 1.5 vvm. When we analyzed the polysaccharide extractions from the cultured mycelium and the culture broth of T. matsutake, 1.4 g of crude polysaccharide was found per 100 g of dried weight in the cultured mycelium, and 1.47 g/l of polysaccharides was found in the culture broth. In addition, the amounts of β-Glucan in the soluble polysaccharide fractions of the cultured mycelium and culture broth were 75.4% and 83.6%, respectively. The cultured mycelium and the culture broth contained a higher amount of β-Glucan than that of the fruiting body.     

Phytochemicals and antioxidant activity of different parts of bambangan (Mangifera pajang) and tarap (Artocarpus odoratissimus)

Mangifera pajang (family: Anacardiaceae; local name: bambangan) and Artocarpus odoratissimus (familiy: Moraceae; local name: tarap) are popular edible fruits in Sabah, Malaysia. The flesh, kernel and peel from M. pajang; seed and flesh from A. odoratissimus were analysed for total antioxidant activity, total polyphenol, total flavonoid and total anthocyanins contents. M. pajang kernel extract displayed the highest free radical scavenging and ferric reducing activities. Total phenolic content of the samples were in the range of 5.96–103.3 mg gallic acid equivalent/g. M. pajang kernel and M. pajang flesh contained the highest and lowest total flavonoid content with the values of 10.98 and 0.07 mg rutin equivalent/g, respectively. The antioxidant activities of extracts were significantly correlated with the total phenolic and flavonoid content (but not the anthocyanins content). The phytochemicals and antioxidant properties of M. pajang and A. odoratissimus, especially their by-products (kernel/seed), indicate that they may impart health benefits when consumed and should be regarded as a valuable source of antioxidant-rich nutraceuticals.     

Antioxidant and radical-scavenging carbazole alkaloids from the oleoresin of curry leaf (Murraya koenigii Spreng.)

Oleoresin of curry leaves (Murraya koenigii Spreng.), obtained using acetone, was evaluated for its antioxidant activity using a β-carotene–linoleic acid model system along with the other extracts obtained using methanol, water and volatile oil. The oleoresin showed maximum activity of 83.2% at 100 ppm among all other extractives in comparison to a synthetic antioxidant, namely, butylated hydroxy anisole which exhibited 90.2% activity at the same concentration. The methanol and water extracts showed activities of 16.7% and 11.3%, respectively, at the same concentration. The volatile oil showed negligible (<10%) activity at 100 ppm concentration. Therefore, the oleoresin was fractionated on a silica gel column to obtain five compounds. Among the five, two compounds with maximum antioxidant activity were identified by ¹H and ¹³C NMR spectra as mahanimbine and koenigine. Koenigine also showed a high degree of radical-scavenging activity.     

Free radical-scavenging and H,K-ATPase inhibition activities of Pithecellobium dulce

The free radical-scavenging property and the inhibitory action on H⁺, K⁺-ATPase activity of aqueous (AEPD) and hydroalcoholic (HAEPD) fruit extracts of Pithecellobium dulce (PD) were screened in various in vitro models. HAEPD showed the greatest free radical-scavenging activity in all the experimental models. Further, HAEPD has shown a potent H⁺, K⁺-ATPase inhibitory activity and comparable to that of Omeprazole, the antiulcer drug. HPLC chromatogram of HAEPD was found to contain rich quantity of phenolic compounds and revealed the presence of flavonoids – quercitrin, rutin, kaempferol, naringin, daidzein. Thus HAEPD was found to possess a good antioxidant capacity when compared to AEPD. The study indicates that PD can be screened for antiulcer activity, since overexpression of H⁺, K⁺-ATPase results in gastric ulceration. So the free radical-scavenging and antiacid secretory activities of PD may be claimed for its traditional consumption for gastric complications.     

Effect of steam treatment on soluble phenolic content and antioxidant activity of the Chaga mushroom (Inonotus obliquus)

The effect of steam treatment on free phenolic acids in Chaga mushrooms (Inonotus obliquus) was investigated. Untreated and steam-treated (120 °C, 3 h) samples of I. obliquus were extracted with organic solvents and free phenolic acid-containing fractions were isolated. Free phenolic acids were determined by LC/PDA (liquid chromatography/photodiode array), ESI LC/MS (electrospray ionisation liquid chromatography/mass spectrometry), and GC/MS (gas chromatography/mass spectrometry). After the steam treatment, the soluble phenolic content determined by modified Folin–Ciocalteu method was increased and antioxidant activity was enhanced, as confirmed by a DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity assay. The amounts of vanillic acid, protocatechuic acid, syringic acid, and 2,5-dihydroxyterephthalic acid were increased significantly as the result of the steam treatment, suggesting that the liberation of low molecular weight free phenolics was enhanced by the steaming process. Consequently, the radical scavenging activity was also significantly enhanced by free phenolics produced using this method.     

Free radical scavenging activity and phenolic content in achenes and thalamus from Fragaria chiloensis ssp. chiloensis, F. vesca and F. x ananassa cv. Chandler

The total phenolic, flavonoid and anthocyanin content of achenes (true fruit) and thalamus (receptacle) from the native South American Fragaria chiloensis ssp. chiloensis (f. patagonica and f. chiloensis), Fragaria vesca and Fragaria x ananassa cv. Chandler was determined by spectrophotometric means. Highest phenolic content was found in F. vesca while lowest content was measured for white strawberry (F. chiloensis ssp. chiloensis, f. chiloensis). The total anthocyanin and total flavonoid contents in the samples investigated was lower for the white strawberry and higher in F. x ananassa cv. Chandler. Total flavonoid content showed a better correlation than total anthocyanins with the free radical scavenging effect of the extracts measured by means of the DPPH discoloration assay. In the superoxide anion assay all the acetone extracts of strawberries showed similar activity. The data presented in this study demonstrate that the amount of phenolic compounds differ significantly between species and subspecies and determine the free radical scavenging activity of fruits. On a w/w basis, higher total phenolics including flavonoids was found in achenes. The highest total anthocyanin content was found in the achenes of F. chiloensis and F. vesca, while F. ananassa presented higher antocyanin content in thalamus. The main anthocyanin in thalamus of F. ananassa (95%) were pelargonidin derivatives which were also present in F. chiloensis ssp. chiloensis f. patagonica (62.6%) but were not detected in F. vesca and F. chiloensis ssp. chiloensis f. chiloensis. In F. vesca and F. chiloensis ssp. chiloensis f. chiloensis only cyanidin derivatives were found both in thalamus and achenes.     

Antioxidant activity of microwave-assisted extract of Buddleia officinalis and its major active component

Buddleia officinalis Maxim, commonly used as rice dye for festivals, was extracted with ethanol using microwave-assisted extraction and Soxhlet extraction. The antioxidant activities of microwave-assisted extract of B. officialis (MEB) and Soxhlet extract of B. officianils (SEB) at the optimum extraction conditions were evaluated and compared with synthetic antioxidant butylated hydroxytoluene (BHT) employing DPPH free radical assay, ABTS assay, total antioxidant activity and reducing power. MEB and SEB had stronger antioxidant activities than BHT in all assays except reducing power, and the effects decreased as follows: MEB > SEB > BHT. The total phenolic contents of MEB and SEB reached 113.56 mg/g and 100.94 mg/g dry weight of extract, respectively, expressed as pyrocatechol equivalents, while the total flavonoids contents were 75.33 mg/g and 62.56 mg/g dry weight of extract, respectively, expressed as catechin equivalents (P < 0.05). Higher phenolic and flavonoids compounds may be major contributors to their higher antioxidant activities. Following activity-oriented separation, luteolin was isolated as an active principle, which exhibited excellent free radical scavenging activities with DPPH IC₅₀ 3.09 μg/ml and ABTS IC₅₀ 2.20 μg/ml.     

Isolation, characterization and immunological activity of a polysaccharide from the fruit bodies of an edible mushroom, Sarcodon aspratus (Berk.) S. Ito.

A water-soluble polysaccharide (HCP) with a molecular mass of 6.7 × 10⁵ Da determined by high performance size-exclusion chromatography (HPSEC), was isolated from the fruit bodies of Sarcodon aspratus (Berk.) S. Ito., an edible mushroom. HCP was elucidated as a liner glucan with a backbone structure of (1 → 6)-linked-??-d-glucopyranosyl residues by interpretation of the composition analysis, methylation analysis, periodate oxidation experiment, FT-IR, and NMR spectroscopy. Immunological activity evaluation using H³-thymidine incorporation method revealed that HCP could significantly stimulate the proliferation of the cultured mice spleen lymphocyte in a dose-dependent manner. HCP might be a potential immunomodulator which can be used against pathogens and tumors in health-care food or medicine. This is the first report on the detailed structure elucidation of the polysaccharide from S. aspratus.     

Antioxidant potentials of buntan pumelo (Citrus grandis Osbeck) and its ethanolic and acetified fermentation products

The antioxidant potentials of buntan pumelo (Citrus grandis Osbeck) and its fermented products were determined. The essential oil from peel had higher total phenolic (214 mM) and flavonoid (134 mg QE/g of dried material) contents than those of different solvent extracts from fruit pulp. In addition, DPPH free radical-scavenging activity and ferric-reducing antioxidant power values determined for the essential oil were 26.1 ± 1.2% and 2.3 ± 0.3 mM, respectively, which were significantly higher than those of various fruit pulp extracts. The ethanol-fermented products of pumelo juice had higher total phenolic and flavonoid contents than those of fresh juice. For maintenance of the substantial antioxidant properties of pumelo products, ethanol-fermented juice rather than fresh or acetate-fermented juice is recommended. Through correlation analysis, the phenolic compounds in the fermented pumelo products were found to be the major contributors to the free radical-scavenging activity and ferric-reducing power.     

Estimation of neuroprotective effects of Laurocerasus officinalis Roem. (cherry laurel) by in vitro methods

Dichloromethane, ethyl acetate, acetone, methanol, and water extracts prepared from the fruits and leaves of Laurocerasus officinalis Roem. (LO) (Rosaceae) were screened for their cholinesterase inhibitory activity against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), the key enzymes in pathogenesis of Alzheimer's disease (AD), using ELISA microplate reader at 50, 100, and 200 ??g mL⁻¹. As AD is associated with oxidative stress, the antioxidant activity of the extracts was also tested by radical-forming methods against 2,2-diphenyl-1-picrylhydrazyl (DPPH), N,N-dimethyl-p-phenylendiamine (DMPD), and superoxide radicals as well as iron-related methods; iron-chelating capacity and ferric-reducing antioxidant power (FRAP) assays. Total phenol and flavonoid quantification was achieved using Folin-Ciocalteau and AlCl₃ reagents, respectively. The highest AChE (44.01 ± 1.75%) and BChE (19.91 ± 0.37%) inhibition was caused by the LO-leaf-methanol extract 200 ??g mL⁻¹, while it showed the best radical-scavenging activity against DPPH at 2000 ??g mL⁻¹. Only, the dichloromethane and water extracts of the fruits and the leaf water extract had an iron-chelating capacity, while the leaf methanol extract displayed the highest FRAP. The leaf methanol extract (113.45 ± 0.71 mg g⁻¹ extract) was found to be the richest in total phenols, while the leaf acetone extract (139.90 ± 4.64 mg g⁻¹ extract) had the most abundant amount of total flavonoids.     

Kinetic modeling of spice extraction from S. aromaticum and C. cassia

A mathematical model was developed for the extracts obtained from Syzygium aromaticum and Cinnamomum cassia with different particle size, solvent–solid ratios on extraction yield. Different particle sizes in the range of 2.8 mm to ?0.5 mm were employed and maximum extraction efficiency was achieved with particles of size ?0.5 mm. Among the solvent–solid ratios (20:1, 30:1, 40:1 and 50:1) ratio of 50:1 showed higher extraction yield. In the extraction kinetics, higher effective diffusivity value of 36.01 × 10⁻¹⁰ m²/s for S. aromaticum and 26.78 × 10⁻¹⁰ m²/s for C. cassia were achieved. Antioxidant values were determined and extracts prepared from ethanol showed higher scavenging activities for S. aromaticum and C. cassia as 78% and 85% respectively. Maximum phenolic content of 1.6 and 12.4 mg GAE/g of sample were achieved for S. aromaticum and C. cassia by hexane and water respectively.     

Free radical scavenging activities and bioactive substances of Jerusalem artichoke (Helianthus tuberosus L.) leaves

The total phenolic content and radical scavenging activities of Jerusalem artichoke (Helianthus tuberosus L.) leaves were investigated. Results indicated that the ethyl acetate fraction contained the highest total phenolic content (266.69 ± 2.51 mg GAE/g dry extract) accompanied with strongest free radical scavenging abilities. Following an in vitro radical scavenging activity-guide fractionation procedure, six phenolic compounds which strongly quenched free radicals were separated from ethyl acetate fraction. Among them, 3-O-caffeoylquinic acid and 1,5-dicaffeoylquinic acid played a dominant role due to their strong free radical scavenging abilities and their high contents. The content of 3-O-caffeoylquinic acid in n-butanol fraction was 74.58 ± 1.05 mg/g, while 1,5-dicaffeoylquinic acid in ethyl acetate fraction was 104.51 ± 2.86 mg/g. The results imply that the leaves of Jerusalem artichoke might be a potential source of natural antioxidants.     

Immunomodulatory and anticancer activities of phenolics from emblica fruit (Phyllanthus emblica L.)

There is a paucity of studies on the immunomodulatory properties of fruit extracts of emblica with the emphasis on lymphocytes. Therefore, the aim of the study was to evaluate the immunomodulatory properties and anticancer potential of six phenolic compounds from emblica fruit by in vitro proliferation assay. Effects of these compounds on splenocyte proliferation and the cytotoxicity to both human breast cancer cell (MCF-7) and human embryonic lung fibroblast cell (HELF) were determined by the MTT method. Significantly stimulatory effects (P < 0.05) were found for geraniin and isocorilagin. The concentration of geraniin, quercetin 3-β-d-glucopyranoside, kaempferol 3-β-d-glucopyranoside, isocorilagin, quercetin, kaempferol and rutin to obtain 50% of stimulatory effect was 56, 123, 242, 42, 73, 93 and 92 μg/ml, respectively. The assay of anticancer activities suggested that geraniin and isocorilagin exhibited higher cytotoxicities than other compounds against MCF-7 with IC₅₀ of 13.2 and 80.9 μg/ml, respectively. Isocorilagin exhibited a strong cytotoxicity to HELF cell with IC₅₀ of 51.4 μg/ml. Geraniin, quercetin, kaempferol and their glycosides had weak cytotoxicity against HELF cells. Paclitaxel showed a strong cytotoxicity to MCF-7 and HELF with IC₅₀ of 6.8 and 14.5 μg/ml, respectively. These findings are in line with the reported potent antioxidant activity. These results suggested that the antitumour activity of these compounds might be achieved by immunomodulatory properties which could partially be attributed to their antioxidant activity.     

Antioxidant activity of cherry laurel fruit (Laurocerasus officinalis Roem.) and its concentrated juice

Cherry laurel fruit and its concentrated juice (pekmez) were examined for their antioxidant activities using different free-radical scavenging activity tests [hydrogen peroxide, superoxide radical, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical], together with reducing power and inhibition of oxidation of human low-density lipoprotein cholesterol. On a fresh weight basis, pekmez exhibited a significantly (P < 0.01) higher antioxidant activity than that of cherry laurel fruit in most cases. However, on a dry weight basis hydrogen peroxide and DPPH radical scavenging activities, and reducing power were significantly higher (P < 0.01) in cherry laurel fruit than in its pekmez, with some exceptions, thus indicating possible destruction of antioxidative compounds during pekmez production. This was also partly due to the moisture content of these two samples. On the basis of the results presented, it is suggested that the intake of cherry laurel fruit and pekmez rich in phenolics would have beneficial effects in improving amelioration of degenerative diseases caused by oxidative stress. Therefore, both cherry laurel fruit and pekmez might be considered as functional food ingredients and nutraceuticals.     

Chemical and antioxidative assessment of dietary turnip (Brassica rapa var. rapa L.)

The phenolic compounds and organic acids of turnip (Brassica rapa var. rapa L.) edible parts (leaves and stems, flower buds and roots) were determined by HPLC–DAD and HPLC–UV, respectively. The results revealed a profile composed of 14 phenolics (3-p-coumaroylquinic, caffeic, ferulic and sinapic acids, kaempferol 3-O-sophoroside-7-O-glucoside, kaempferol 3-O-sophoroside-7-O-sophoroside, kaempferol 3-O-(feruloyl/caffeoyl)-sophoroside-7-O-glucoside, kaempferol 3,7-O-diglucoside, isorhamnetin 3,7-O-diglucoside, kaempferol 3-O-sophoroside, 1,2-disinapoylgentiobiose, 1,2′-disinapoyl-2-feruloylgentiobiose, kaempferol 3-O-glucoside and isorhamnetin 3-O-glucoside) and six organic acids (aconitic, citric, ketoglutaric, malic, shikimic and fumaric acids). The quantification of the identified compounds showed kaempferol 3-O-sophoroside-7-O-glucoside, kaempferol 3-O-(feruloyl/caffeoyl)-sophoroside-7-O-glucoside, isorhamnetin 3,7-O-diglucoside and isorhamnetin 3-O-glucoside as the main phenolics, and malic acid as the organic acid present in highest amounts. A screening of the antioxidative potential was also performed by means of the DPPH radical scavenging assay. Turnip flower buds exhibited the strongest antioxidant capacity.