Physico-chemical characterization, antioxidant and anticancer activities in vitro of a novel polysaccharide from Melia toosendan Sieb. Et Zucc fruit

A novel water-soluble polysaccharide pMTPS-3, obtained from Melia toosendan Sieb. Et Zucc fruit by hot-water extraction and ethanol precipitation, was fractionated by DEAE-52 cellulose anion-exchange and Sephadex G-100 gel filtration chromatography. Its primary structural features and molecular weight were characterized by Fourier infrared spectrometry (FTIR), gel permeation chromatography (GPC) and gas chromatography (GC). And the antioxidant activities of pMTPS-3 in vitro were evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, superoxide radical scavenging assay and hydroxyl radical scavenging assay. The results suggested that pMTPS-3 was a heteropolysaccharide, composed of arabinose, glucose, mannose, and galactose in the molar ratio of 17.3:28.3:41.6:12.6 with molecular weight 26 100 Da. The purified pMTPS-3 was revealed to have notable scavenging activity against DPPH radical in a concentration-dependent manner and present a moderate inhibition of superoxide radicals with an IC₅₀ (5.6 mg/ml), and potent inhibiting power for hydroxyl radical compared with crude polysaccharide. Further, it exhibited strong inhibition effect in vitro on the growth of human gastric cancer BGC-823 cells. It is strongly evidenced that pMTPS-3 purified from the crude polysaccharides of Melia toosendan Sieb. Et Zucc could be explored as a potential antioxidant and therapeutics.     

Structure and antioxidant activity of an extracellular polysaccharide from coral-associated fungus, Aspergillus versicolor LCJ-5-4

An extracellular polysaccharide AVP was isolated from the fermented broth of coral-associated fungus Aspergillus versicolor LCJ-5-4. AVP was a mannoglucan with molecular weight of about 7 kDa, and the molar ratio of glucose and mannose was 1.7:1.0. On the basis of detailed one- and two-dimensional nuclear magnetic resonance (1D and 2D NMR) spectroscopic analyses, the backbone of AVP was characterized to be composed of (1 → 6)-linked α-d-glucopyranose and (1 → 2)-linked α-d-mannopyranose units. The mannopyranose residues in the backbone were substituted mainly at C-6 by the side chain of (1 → 2)-linked α-d-mannopyranose trisaccharides units. The antioxidant activity of AVP was evaluated with the scavenging abilities on 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide and hydroxyl radicals in vitro, and the results indicated that AVP had good antioxidant activity, especially scavenging ability on superoxide radicals. AVP was a novel extracellular polysaccharide with different structural characteristics from other extracellular polysaccharides and could be a potential source of antioxidant.     

Chemical composition and bioactivities of a water-soluble polysaccharide from the endodermis of shaddock

The chemical composition of shaddock (Citrus paradisi) mainly consisted of polyphenols, proteins and polysaccharides. However, polysaccharides from shaddock materials have received much less consideration than polyphenols. Herein, a water-soluble neutral polysaccharide from the endodermis of shaddock was isolated and showed good bioactivities. Crude polysaccharides from the endodermis of shaddock (EPS) was extracted with hot water and separated on a DEAE Sepharose FF gel filtration column to obtain NEPS. The IR and UV spectra of NEPS showed that NEPS was mainly composed of polysaccharide and there are no proteins existing in NEPS. The DPPH radical scavenging and reducing power of NEPS are much lower than those of crude EPS; however, Citrus flavonoids significantly improved the DPPH radical scavenging potential and reducing power of NEPS. The crude EPS (5mg/mL) showed a similar inhibitory effect (77.92±5.03%) with NEPS (5mg/mL) (74.63±4.71%) on α-amylase.     

Antioxidant activities potential of tea polysaccharide fractions obtained by ultra filtration

Three polysaccharide fractions (TPS1, TPS2 and TPS3) with different molecular weights were obtained using ultra filtration membranes from crude tea polysaccharide (CTPS) extracted from abandoned lower grade tea leaves. Each fraction contained different contents of neutral sugar, uronic acid, protein, and total polyphenols. These differences provided basis for the antioxidant and free radical scavenging activity of these polysaccharide fractions. The molecular weights of TPS1, TPS2, and TPS3 were around 2.40×10(5) Da, 2.14×10(4) Da, and 2.46×10(3) Da, respectively. In general, TPS1 and CTPS had stronger antioxidant activity, TPS2 and TPS3 had lower antioxidant activity. TPS1 had higher activity for DPPH and lipid per oxidation inhibition. But it had lower capacity for reducing power and metal chelating. This might be due to its higher content of hexuronic acid and larger molecular weight. The order of inhibition activity of lipid per oxidation of various polysaccharide fractions was the same as DPPH radical scavenging activity, as well as the order of metal chelating activity of various polysaccharide fractions similar to hydroxyl radical scavenging activity, which demonstrated that hydroxyl radical scavenging activity of polysaccharide relied heavily on the Fe(2+) metal chelating to decrease the generation of hydroxyl radical.     

藜叶中黄酮类化合物体外抗氧化活性研究

为了探讨藜叶中黄酮类化合物的体外抗氧化活性,采用有机溶剂提取法和色谱柱法对藜叶中化学成分进行提取与分离;以Vc作对照,对分离纯化的芦丁、乙酸乙酯浸膏和正丁醇浸膏进行DPPH·、O2^-·和·OH的清除效果试验。结果表明：三者对DPPH·、O2^-·、·OH均具有清除作用,且与浓度呈量效关系,芦丁对·OH清除效果优于Vc,芦丁具有较强的清除DPPH·能力,其IC50为0．05μg／mL。     

Extraction of polysaccharides and the antioxidant activity from the seeds of Plantago asiatica L

The extraction conditions of polysaccharides from Plantago asiatica L. seeds were investigated. Four parameters affecting the polysaccharides extraction, extraction times, water to sample, extraction temperature and single extraction time, were determined by orthogonal experiments. Under the optimized conditions, the polysaccharides yield of P. asiatica L. seeds was 2.467%. The antioxidant activities of the polysaccharides were investigated. The reducing power of the polysaccharides was dose dependent, and the reducing capacity of the polysaccharides was inferior to butylated hydroxytoluene, which is known to be a strong reducing agent. The scavenging rates of the polysaccharides on superoxide and 1,1-diphenyl-2-picrylhydrazyl radicals were79.7% and 81.4%, at polysaccharides concentration of 0.75 mg/mL, respectively, a scavenging rates approximately similar to that of 0.75 mg/mL ascorbic acid (83.5% and 85.1%, respectively). Furthermore, it exhibited a moderate concentration-dependent ABTS radical scavenging activity, ferrous ion chelating potency and H₂O₂ scavenging activity. The data obtained in the in vitro models clearly establish the antioxidant potency of the polysaccharides extracted from Semen Plantaginis.     

荸荠皮多糖体外清除自由基活性的研究

采用DPPH自由基法、邻苯三酚自氧化法和水杨酸法分别表征了荸荠皮多糖清除DPPH自由基(DPPH.)、超氧阴离子自由基(O2-.)和羟基自由基(.OH)的能力。结果显示,脱蛋白和未脱蛋白荸荠皮多糖具有清除自由基的活性,但该活性明显低于茶多酚。脱蛋白多糖清除DPPH.的能力高于未脱蛋白多糖,其清除50%DPPH.的作用质量浓度(EC50)分别为50.26 mg.L-1和76.22 mg.L-1。两种多糖清除.OH和O2-.的能力相当,其EC50(g.L-1)分别为0.118和0.124以及10.87和9.53。     

Characterization and application of a detergent-stable alkaline α-amylase from Bacillus subtilis strain AS-S01a

A strain AS-S01a, capable of producing high-titer alkaline α-amylase, was isolated from a soil sample of Assam, India and was taxonomically identified as Bacillus subtilis strain AS-S01a. Optimized α-amylase yield by response surface method (RSM) was obtained as 799.0 U with a specific activity of 201.0 U/mg in a process control bioreactor. A 21.0 kDa alkaline α-amylase purified from this strain showed optimum activity at 55 °C and pH 9.0, and it produced high molecular weight oligosaccharides including small amount of glucose from starch as the end product. The K_m and V_max values for this enzyme towards starch were determined as 1.9 mg/ml and 198.21 μmol/min/mg, respectively. The purified α-amylase retained its activity in presence of oxidant, surfactants, EDTA and various commercial laundry detergents, thus advocating its suitability for various industrial applications.     

Antioxidant activity of a water-soluble polysaccharide purified from Pteridium aquilinum

A water-soluble crude polysaccharide, obtained from fern Pteridium aquilinum, was fractionated by DEAE-Sepharose Fast-Flow column chromatography, and purified by Sephacryl S-400 HR column chromatography. The average molecular weight (M_w) of the purified polysaccharide (PLP) is 458,000 Da. The monosaccharide components of PLP were characterized by gas chromatography (GC), and the majority of the monosaccharide components was glucose (relative mass 58.1%) with low levels of galactose, mannose, rhamnose, and arabinose (relative mass 18.7%, 6.8%, 10.2%, and 6.1%, respectively). The Fourier-transform infrared spectra (FTIR) of PLP revealed typical characteristics of polysaccharides. On the basis of the ferric-reducing antioxidant power assay (FRAP), DPPH radical-scavenging, the superoxide radical assay, and self-oxidation of 1,2,3-phentriol assay, the antioxidant activities of PLP were investigated. The purified polysaccharide was demonstrated to have strong reductive power (FRAP value: 827.6 μmol/L), moderate scavenging activities against DPPH radicals (83.1%) and superoxide radicals (60.5%), and moderate inhibiting power for self-oxidation of 1,2,3-phentriol (52.4%).     

不同提取方法对豫产白花蛇舌草多糖及抗氧化活性的影响

为评价不同提取方法对河南地区白花蛇舌草粗多糖的得率、总碳水化合物含量、杂质含量和抗氧化活性的影响,本实验分别以传统热水提取法、超声波辅助提取法、纤维素酶提取法从该药材中提取多糖。同时测定了粗多糖中总碳水化合物含量、杂质含量(总蛋白和总酚含量)以及羟基自由基清除能力、DPPH自由基清能力、还原力等抗氧化能力。实验结果显示三种提取法所得粗多糖相对于原药材的得率分别为6.77±0.09%、6.37±0.01%和4.81±0.90%;粗多糖中的总碳水化合物含量分别为289.14±1.33、246.83±2.33和278.92±2.92mg/g。粗多糖中总蛋白含量分别为28.59±2.21、23.26±2.43和4.96±0.18mg/g,总酚类成分含量分别为5.9±0.08、5.31±0.40和2.82±0.07mg/g。抗氧化活性实验结果表明,三种粗多糖的羟基自由基清除能力呈现出浓度依赖性,且均在3.0mg/mL时达到最大,对应最大清除率分别为89.13%、85.87%和74.71%。另外,三种粗多糖的DPPH自由基清除率分别在0.75~3.0mg/mL的浓度区间内达到最大并保持稳定,对应清除率的稳定区间分别为77.02%~77.90%、77.06%~77.96%和83.16%~85.73%。综上所述,以上三种不同提取方法对白花蛇舌草多糖的品质影响较大,可以为豫产白花蛇舌草的品质鉴别、多糖质量控制和开发利用提供依据。     

滑子菇多糖的体外生物活性初步研究

为了研究滑子菇水提粗多糖(PNP)的体外生物活性,对滑子菇多糖的总还原力、清除1,1-二苯基-2-苦苯肼自由基(DPPH·)和由Fe2+诱发的脂质过氧化反应的抑制作用进行研究,采用MTT比色法和胎盘蓝细胞计数检测对滑子菇水提粗多糖的体外抑制K562细胞生长作用进行了研究,采用流失细胞术对滑子菇多糖作用人白血病K562细胞后的细胞周期进行了研究。结果表明:滑子菇水提粗多糖PNP具有一定的还原能力;在高质量浓度(800μg/mL)时具有接近于Vc清除DPPH·的能力,达41.28%;PNP对Fe2+诱发的脂质过氧化反应具有一定的抑制作用,并且随着浓度的增加抑制作用逐渐增强,但总的增长趋势不大;MTT实验表明PNP对K562细胞的体外增长有抑制作用,在质量浓度为800μg/mL和作用时间为48 h时,可达到最高的抑制率35.03%。流式细胞术对细胞周期的检测表明滑子菇多糖能够阻滞人白血病K562细胞于G1期。     

不同炮制方法对玉竹提取物得率及体外抗氧化作用的影响

以多糖、总黄酮、醇溶物和水溶物的得率及体外抗氧化活性为考察指标,研究了酒蒸和蜜蒸两种炮制方法对玉竹品质的影响。结果表明：酒蒸炮制玉竹的多糖、醇溶物得率最高,蜜蒸炮制玉竹总黄酮、水溶物的得率最高,比未炮制的玉竹（生品玉竹）中相应成分的得率分别提高了43.86%、29.53%、49.46%和34.66%。将多糖、水溶性浸出物、醇溶性浸出物及总黄酮四者得率相加的和进行比较,蜜蒸最好,蜜蒸为111.069%,酒蒸为107.309%,生品玉竹为80.926%。酒蒸炮制玉竹的多糖、水溶物对DPPH自由基的清除率均高于蜜蒸玉竹和生品玉竹,其DPPH_IC50分别为0.345±0.019和0.441±0.022 mg·mL^-1;蜜蒸炮制玉竹的总黄酮、醇溶物对DPPH自由基的清除率均高于酒蒸玉竹和生品玉竹,其DPPH_IC50分别为0.047±0.011和0.199±0.036 mg·mL^-1;在浓度为1 mg·mL^-1时,蜜蒸玉竹总黄酮对DPPH自由基的清除率最大,为90.29%,超过了浓度为0.05 mg·mL^-1的芦丁和槲皮素标品对DPPH自由基的清除率。两种炮制方法均提高了多糖、水溶物、总黄酮3种提取物的还原能力,但是降低了醇溶物的还原能力。     

Purification and biochemical characterization of antioxidant peptide from horse mackerel (Magalaspis cordyla) viscera protein

In the present study, a peptide having high antioxidant properties was isolated from horse mackerel viscera protein, Magalaspis cordyla. In vitro gastrointestinal digestion was employed to obtain potential protein hydrolysate and was subjected to consecutive chromatographic methods using fast protein liquid chromatography (FPLC) connected to diethyl amino ethyl (DEAE) anion exchange column and Sephadex G-25 gel filtration column. The activity of the fractions was tested against DPPH and hydroxyl radicals and the isolated peptide showed 89.2 and 59.1 percentage of scavenging. The amino acid sequence of purified peptide was determined using ESI-MS/MS as Ala-Cys-Phe-Leu (518.5 Da), it exhibited high activity against polyunsaturated fatty acid (PUFA) peroxidation than that of natural antioxidant, α-tocopherol.     

Chemical modification,antioxidant and α-amylase inhibitory activities of corn silk polysaccharides

Water-soluble corn silk polysaccharides (CSPS) were chemically modified to obtain their sulfated, acetylated and carboxymethylated derivatives. Chemical characterization and bioactivities of CSPS and its derivatives were comparatively investigated by chemical methods, gas chromatography, gel filtration chromatography, scanning electron microscope, infrared spectroscopy and circular dichroism spectroscopy, scavenging DPPH free radical assay, scavenging hydroxyl radical assay, ferric reducing power assay, lipid peroxidation inhibition assay and α-amylase activity inhibitory assay, respectively. Among the three derivatives, carboxylmethylated polysaccharide (C-CSPS) demonstrated higher solubility, narrower molecular weight distribution, lower intrinsic viscosity, a hyperbranched conformation, significantly higher antioxidant and α-amylase inhibitory abilities compared with the native polysaccharide and other derivatives. C-CSPS might be used as a novel nutraceutical agent for human consumption.     

Antioxidant, α-glucosidase inhibitory and anti-inflammatory effects of aerial parts extract from Korean crowberry (Empetrum nigrum var. japonicum)

Crowberry (Empetrum nigrum L.) is a wild berry commonly found in the northern hemisphere. Crowberry fruits have been suggested as good resources for functional applications in the cosmetic and pharmaceutical industries, but the high polyphenolic content in crowberry leaves also indicates crowberry aerial parts as potential dietary health supplements. In this study, therefore, the biological activities of the aerial parts of Korean crowberry (E. nigrum var. japonicum) were investigated. Antioxidant activity was measured by three different assays on DPPH free radical scavenging, reducing power, and total antioxidant capacities. Dose-dependent antioxidant activities were exhibited by crude methanol extract and its fractions, suggesting that the crude methanol extract and EtOAc fraction possessed strong antioxidant activities and capacities. In addition, the crude methanol extract and EtOAc strongly inhibited α-glucosidase activity and suppressed the secretion of pro-inflammatory mediator and nitrite oxide from LPS-stimulated RAW 264.7 cells. These findings provide valuable evidence for the potential of such parts as good dietary sources of natural antioxidant, α-glucosidase inhibitory, and anti-inflammatory components, suggesting that using the non-edible parts (e.g., leaves and stems) of crowberry can be a potential natural avenue for improving human health.     

桦褐孔菌活性物质的提取工艺及体外抗糖尿病活性

对桦褐孔菌活性物质的提取工艺及其体外抗糖尿病活性进行研究。桦褐孔菌子实体用乙醇浸提后,乙醇粗提物用不同有机溶剂萃取,醇提残渣再以热水浸提,用标准曲线法测定活性组分中活性成分的含量,并检测活性物质对羟基自由基(·OH)、超氧阴离子自由基(O2-·)的清除效果以及对关键糖代谢酶α-淀粉酶、α-葡萄糖苷酶的抑制作用。结果表明:4种活性组分(乙酸乙酯相、正丁醇相、水相和粗多糖)对羟基自由基(·OH)和超氧阴离子自由基(O2-·)都有较强的清除作用,其中乙酸乙酯组分的活性最高,乙醇粗提物萃取组分对α-淀粉酶有抑制活性,而粗多糖对α-葡萄糖苷酶有抑制作用;桦褐孔菌具有抗氧化和抗糖尿病活性,其活性与活性物质种类及其含量具有相关性。     

Contribution of phenolic compounds to the antioxidant potential and type II diabetes related enzyme inhibition properties of Pongamia pinnata L. Pierre seeds

The methanolic extract of Pongamia pinnata L. Pierre (locally called as karanja) seed materials, an underutilized food legume collected from India was analyzed for antioxidant and type II diabetes related enzyme inhibition properties. The methanolic extract of raw seeds contained total free phenolic content of 14.85 ± 0.32 g catechin equivalent/100 g extract DM. Encouraging levels of ferric reducing/antioxidant power (FRAP, 1179 mmol Fe[II]/mg extract), inhibition of β-carotene degradation (41.13%) and radical scavenging activity against DPPH (54.64%) and superoxide (54.53%) were exhibited by the raw sample. Further, it also recorded 77.92% of α-amylase and 86.50% of α-glucosidase enzyme inhibition characteristics under in vitro starch digestion bioassay. Sprouting + oil-frying caused a apparent increase on the total free phenolic content and also significant improvement on the antioxidant and free radical scavenging capacity of P. pinnata seeds, while soaking + cooking as well as open-pan roasting treatments showed diminishing effects. Moreover, inhibition of α-amylase and α-glucosidase enzyme activities was declined to 24.24 and 45.14%, respectively during sprouting + oil-frying treatment, which are more desirable for the dietary management of type II diabetic patients.     

Bio-guided isolation of potential antimicrobial and antioxidant agents from the stem bark of Trilepisium madagascariense

Aim

This study describes the activity-guided isolation of antimicrobial and antioxidant agents from Trilepisium madagascariense stem bark.

Methods

The methanol crude extract of T. madagascariense was partitioned sequentially into n-hexane, ethyl acetate, n-butanol and the residual aqueous fractions. The ethyl acetate fraction was subjected to column chromatography and the structures of isolated compounds were elucidated using GC-MS and/or NMR data by comparing with those reported in the literature. Antimicrobial activity was assayed by agar well diffusion and broth microdilution techniques on 8 bacteria and 10 yeasts. The antioxidant activity was determined by DPPH radical scavenging method.

Results

The bioassay-guided fractionation of the crude methanol extract of T. madagascariense afforded two known compounds [vanillic acid () and isoliquiritigenin ()] and two mixtures of fatty acids (n-hexane fraction and first column fraction of ethyl acetate fraction, F1). The fractionation of the crude methanol extract enhanced the antimicrobial activity. Compound 2 was generally more active than compound 1. For all the tested samples, the most sensitive microbes were Enterococcus faecalis ATCC 10541 (MIC range of 60-780 μg/ml) for bacteria and Candida guillermondi (MIC range of 0.01-190 μg/ml) for yeasts. The DPPH radical scavenging activity (RSa) of compound 2 (RSa₅₀ = 28.73 μg/ml) was comparable to that of the crude methanol extract (RSa₅₀ = 29.92 μg/ml).

Conclusion

The antimicrobial activities and the antioxidant properties of the methanol crude extract, fractions and compounds 1 and 2 from the stem bark of T. madagascariense are being reported for the first time. These results may justify the traditional use of this plant for the treatment of gastrointestinal disorders.     

Carboxymethylation of polysaccharides from Auricularia auricula and their antioxidant activities in vitro

The chemical carboxymethylated polysaccharide (CMAAP), which is derived from the poorly water-soluble, crude polysaccharide from Auricularia auricula (AAP), was prepared. The degree of substitution of CMAAP was 0.857, and its solubility in water reached 0.6 mg/mL, much higher than that of AAP (0.1 mg/mL). One fraction, CMAAP22, was purified. Chemical analysis indicated that CMAAP22 was composed of mannose and glucose in a ratio of 1.06:1 with a molecular weight of 3.4 × 10⁶ Da. IR spectra and ¹³C NMR spectra indicated that the C2, C4 and C6 positions were partially substituted by -CH₂COOH. AFM revealed that CMAAP22 was spherical particle structure with a diameter of 80-300 nm. Moreover, the antioxidant activity of CMAAP22 was improved, nearly twice as much as that of the AAP, especially in the scavenging of DPPH and ABTS+.