Constituents of the seed of Malva verticillata. VIII. Smith degradation of MVS-VI, the major acidic polysaccharide, and anti-complementary activity of products.

The controlled Smith degradation of MVS-VI, the major acidic polysaccharide having remarkable anti-complementary activity isolated from the seeds of Malva verticillata L., was performed. Methylation analysis of both the primary and the secondary Smith degradation products indicated that the core structural features of MVS-VI include a backbone chain composed of beta-1,3-linked D-galactose residues. The majority of galactose units in the backbone carry side chains composed of beta-1,3- and beta-1,6-linked D-galactosyl residues at position 6. The controlled Smith degradation products showed considerable anti-complementary activity.     

Mode of complement activation by acidic heteroglycans from the leaves of Artemisia princeps PAMP.

The mode of action of the anti-complementary acidic heteroglycans, AAF-IIb-2 and IIb-3 which consisted of rhamnogalacturonan core and arabinogalactan moieties, purified from the leaves of Artemisia princeps PAMP (Japanese name = Gaiyo) were investigated. The anti-complementary activities of AAF-IIb-2 and IIb-3 were reduced partially in the absence of Ca2+ ions. A marked consumption of C4 was observed to have occurred when serum was incubated with both polysaccharides in the presence of Ca2+ ions. AAF-IIb-2 showed more potent C4 consumption than IIb-3. After the incubation of the serum with AAF-IIb-2 in the absence of Ca2+ ions, a cleavage of C3 in the serum was detected by immunoelectrophoresis. AAF-IIb-2 showed more significant consumption of the complement than IIb-3 when rabbit erythrocytes were used in the assay system in the absence of Ca2+ ions. These results indicate that AAF-IIb-2 activates the complement via both the alternative and classical pathways, whereas IIb-3 mainly activates the complement via the classical pathway. The absorption of serum with Protein A-Sepharose results in a decrease of the activity of AAF-IIb-2 and IIb-3. However, the decrease of the activity was restored by the replacement of the immunoglobulin G (IgG) fraction after its recovery from the Protein A-Sepharose. These results suggest that IgG dependent mechanisms are both involved in the anti-complementary activity of AAF-IIb-2 and IIb-3.     

Constituents of the seed of Malva verticillata. VI. Characterization and immunological activities of a novel acidic polysaccharide

A novel acidic polysaccharide, designated as MVS-VI, was isolated from the seeds of Malva verticillata L. It was homogeneous on electrophoresis and gel chromatography, and its molecular mass was estimated to be 26000. It is composed of L-arabinose: D-xylose: D-galactose: D-glucose: L-rhamnose: D-galacturonic acid in the molar ratio of 30: 15: 20: 3: 2: 10, in addition to small amounts of peptide moiety. Methylation analysis, carbon-13 nuclear magnetic resonance and periodate oxidation studies indicated its structural features to have mainly acidic alpha-arabino-3,6-beta-galactan type structural units. MVS-VI showed significant reticuloendothelial system-potentiating activity in a carbon clearance test, and it possesses remarkable anti-complementary activity.     

Plant mucilages. XLII. An anti-complementary mucilage from the leaves of Malva sylvestris var. mauritiana

A mucilage, designated as MSL-M, was isolated from the leaves of Malva sylvestris L. var. mauritiana Mill. It was homogeneous on electrophoresis and gel chromatography. Its intrinsic viscosity value in aqueous solution was 12.0, and its molecular weight was estimated to be about 6.0 x 10(6). The major constituent is an acidic polysaccharide composed of L-rhamnose: D-galactose: D-galacturonic acid: D-glucuronic acid in the molar ratio of 6:3:2:2. Methylation analysis of both the mucilage and the carboxyl-reduced derivative, carbon-13 nuclear magnetic resonance and partial hydrolysis studies indicated its main structural features. It has considerable anti-complementary activity.     

The core structure and immunological activities of glycyrrhizan UA, the main polysaccharide from the root of Glycyrrhiza uralensis.

The controlled Smith degradation and limited hydrolysis of glycyrrhizan UA, the main phagocytosis-activating polysaccharide isolated from the root of Glycyrrhiza uralensis FISCHER, was performed. The reticuloendothelial system-potentiating, anti-complementary and alkaline phosphatase-inducing activities of glycyrrhizan UA and its degradation products were investigated. Methylation analyses of primary, secondary and tertiary Smith degradation products and of the limited hydrolysis product indicated that the core structural features of glycyrrhizan UA include a backbone chain composed of beta-1,3-linked D-galactose. All of the galactose units in the backbone carry side chains composed of mainly alpha-1,5-linked L-arabino-beta-1,6- or 1,3-linked D-galactose residues at position 6. Removal of the arabinosyl side chains caused a pronounced decrease in immunological activity.     

关白附中1,6-葡聚糖及其硫酸酯的抗补体活性

为研究关白附多糖及其硫酸酯的经典途径抗补体活性, 以关白附[Aconitum coreanum(Lévl.) Raipaics]为原料, 经水提醇沉、 DE-32、 Superdex-200和Superdex-75凝胶柱分离纯化, 得到1个均一的中性多糖KMPS-2A. 采用高效凝胶渗透色谱法、 甲基化、 核磁共振和红外光谱等手段对KMPS-2A的结构进行了鉴定; 采用氯磺酸吡啶法制备了多糖硫酸酯, 并测定了多糖及硫酸酯的抗补体活性. 结果表明, KMPS-2A的平均相对分子量为6.76×10⁵, 结构为α-1,6-D-Glc链接的线性多糖; 在氯磺酸与吡啶的体积比为1.75∶1.0时制备的多糖硫酸酯1.75B的取代度最高为1.79. 碳核磁谱分析结果表明, 硫酸基团先后取代C2, C3及C4位. 该多糖硫酸酯的抗补体活性与其硫酸基团取代度呈现一定的相关性, 多糖硫酸酯1.75B的经典途径抗补体活性优于阳性对照药肝素, 表明其具有开发成为补体抑制剂的潜力.     

Identification of potential antioxidant compounds in the essential oil of thyme by gas chromatography with mass spectrometry and multivariate calibration techniques

Thyme species are used in traditional medicine throughout the world and are known for their antiseptic, antispasmodic, and antitussive properties. Also, antioxidant activity is one of the interesting properties of thyme essential oil. In this research, we aim to identify peaks potentially responsible for the antioxidant activity of thyme oil from chromatographic fingerprints. Therefore, the chemical compositions of hydrodistilled essential oil of thyme species from different regions were analyzed by gas chromatography with mass spectrometry and antioxidant activities of essential oils were measured by a 1,1‐diphenyl‐2‐picrylhydrazyl radical scavenging test. Several linear multivariate calibration techniques with different preprocessing methods were applied to the chromatograms of thyme essential oils to indicate the peaks responsible for the antioxidant activity. These techniques were applied on data both before and after alignment of chromatograms with correlation optimized warping. In this study, orthogonal projection to latent structures model was found to be a good technique to indicate the potential antioxidant active compounds in the thyme oil due to its simplicity and repeatability.     

Isolation,Structural Elucidation,Antioxidant and Hypoglycemic Activity of Polysaccharides of Brassica rapa L.

The aim of this study was to investigate the effects of microwave ultrasonic-assisted extraction (MUAE) on the content, structure, and biological functions of Brassica rapa L. polysaccharide (BRP). Response surface methodology (RSM) was used to optimize the parameters of MUAE, and it obtained a polysaccharide with yield of 21.802%. Then, a neutral polysaccharide named BRP-1-1 with a molecular weight of 31.378 kDa was isolated and purified from BRP using DEAE-650 M and Sephadex G-100. The structures of the BRP-1-1 were elucidated through a combination of FT-IR, GC-MS, NMR, and methylation analysis. The results showed that BRP-1 consisted of mannose (Man) and glucose (Glu) in a molar ratio of 7.62:1. The backbone of BRP-1-1 mainly consisted of →6)-α-D-Glup-(1→4-β-D-Glup-(1→2)-α-D-Manp-(1→2)-α-D-Glup-(1→, the branch was [T-α-D-Manp-(1]_n→. BRP-1-1 intervention significantly inhibited α-glucosidase activity; an inhibition rate of 44.623% was achieved at a concentration of 0.5 mg/mL. The results of the in vitro biological activity showed that BRP-1-1 has good antioxidant and hypoglycemic activity, suggesting that BRP-1-1 could be developed as a functional medicine.     

The core structure of ukonan A, a phagocytosis-activating polysaccharide from the rhizome of Curcuma longa, and immunological activities of degradation products.

The controlled Smith degradation of ukonan A, a phagocytosis-activating polysaccharide isolated from the rhizome of Curcuma longa L., was performed. The reticuloendothelial system-potentiating, anti-complementary and alkaline phosphatase-inducing activities of ukonan A and its degradation products were investigated. Methylation analyses of both the primary and the secondary Smith degradation products indicated that the core structural features of ukonan A include a backbone chain mainly composed of beta-1,3-linked D-galactose, beta-1,4-linked D-xylose and alpha-1,2-linked L-rhamnose residues. All of the galactose units in the backbone carry side chains composed of alpha-L-arabino-beta-D-galactosyl or beta-D-galactosyl residues at position 6. Ukonan A has a remarkable effect on each of the three kinds of immunological activities. Periodate oxidation caused pronounced decrease or disappearance of the activities, but the controlled Smith degradation product having the core structure of polysaccharide showed considerable restoration of these activities.     

Purification, characterization and antioxidant activities in vitro and in vivo of the polysaccharides from Boletus edulis bull

A water-soluble polysaccharide (BEBP) was extracted from Boletus edulis Bull using hot water extraction followed by ethanol precipitation. The polysaccharide BEBP was further purified by chromatography on a DEAE-cellulose column, giving three major polysaccharide fractions termed BEBP-1, BEBP-2 and BEBP-3. In the next experiment, the average molecular weight (Mw), IR and monosaccharide compositional analysis of the three polysaccharide fractions were determined. The evaluation of antioxidant activities both in vitro and in vivo suggested that BEBP-3 had good potential antioxidant activity, and should be explored as a novel potential antioxidant.     

An acidic polysaccharide having immunological activities from the rhizome of Cnidium officinale.

An acidic polysaccharide, designated as cnidirhan AG, was isolated from the rhizomes of Cnidium officinale Makino. It was homogeneous on electrophoresis and gel chromatography, and its molecular mass was estimated to be 5.1 x 10(4). It showed pronounced reticuloendothelial system-potentiating activity in a carbon clearance test, and had a remarkable effect on both anti-complementary and alkaline phosphatase-inducing activities. It is composed of L-arabinose: D-galactose: D-glucuronic acid in the molar ratio of 2:6:1, in addition to small amounts of O-acetyl groups. Methylation analysis, carbon-13 nuclear magnetic resonance, controlled Smith degradation and limited acid hydrolysis indicated that the core structural features of cnidirhan AG include a backbone chain composed of beta-1,3-linked D-galactose residues. Some of the galactose units in the backbone carry beta-D-galactosyl side chains at position 6. Both alpha-L-arabinosyl arabinose side chains and terminal beta-D-glucuronic acid residues are linked to the core galactan units.     

Structural Characterization and Antioxidative Activity of Polysaccharide from Adenophora Potaninii

The bioactivity of polysaccharides extracted from Adenophora Potaninii have been investigated^[1,2], but its chemical structure has not been reported. We now describe the structure and the antioxidation activity of a water soluble polysaccharide, AP_l, which was extracted from the roots of Adenophora Potaninii by boiling water. The crude polysaccharide was isolated and purified by DEAE-cellulose and Sephadex G-200 column, thus, AP₁ white powders, was obtained.     

Characterization of a kerationlytic metalloprotease from Bacillus sp. SCB-3

A keratinolytic protease-producing microorganism was isolated from soybean paste waste and was identified as a strain of Bacillus sp. The keratinase was purified by polyethylene glycol precipitation and two successive column chromatographies with DEAE-Toyopearl 650C and Sephacryl S-200 HR. The purified enzyme had overall 11 purification folds with an 18% yield. The results of sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel filtration on Sephacryl G-200 indicated that the purified enzyme was monomeric and had a molecular weight of 134 kDa. The optimum temperature and pH were 40°C and 7.0, respectively. This enzyme was completely inhibited by EDTA and EGTA, and it was restored by the addition of Ca⁺² and Mg⁺². These results suggested that it is a metalloprotease. The stimulated enzyme activity by reducing agents indicated that the reducing condition was important in the expression of the activity.     

普鲁兰多糖的分离纯化及结构鉴定

采用Ｓａｖａｇｅ法，ＤＥＡＥ－纤维素离子交换层析法及Ｓｅｐｈａｄｅｘ Ｇ－１００凝胶过滤法提纯普鲁兰多糖。纯化的普鲁兰多糖的结构经过薄层层析，红外光谱及核磁共振光谱加以鉴定。     

Purification and characterization of an extracellular lipolytic enzyme from the fermented fish-originated halotolerant bacterium, <Emphasis Type="Italic">Virgibacillus alimentarius</Emphasis> LBU20907

A halotolerant Virgibacillus alimentarius LBU20907 isolated from fermented fish (Budu) was found to be an efficient producer of extracellular halophilic lipase enzyme. The enzyme was purified 5.99-fold with a 0.15% final yield to homogeneity by ammonium sulfate precipitation, followed by dialysis, Toyopearl DEAE-650 M ion exchange chromatography, Toyopearl butyl-650 M hydrophobic interaction chromatography, and Toyopearl-HW 55 F gel filtration chromatography. SDS-PAGE of purified lipase exhibited a homogenous single band with a very high molecular weight of 100 kDa. The properties of purified lipase revealed maximum activity at pH 7.0 and 40 °C. It was also highly stable in a pH range of 6.0–7.0, retaining more than 90% activity for 24 h. It was stable at the temperature of 30–50 °C and maintained more than 80% activity for 16 h. The purified lipase performing the maximal activity in the presence of 20.0% NaCl indicated halophilic enzyme properties. Its lipolytic activity was highest against p-nitrophenyl palmitate. The lipase activity was found to be enhanced in hexane. The enzyme activity was stimulated in the presence of Zn²⁺, Ca²⁺, Mg²⁺, and Sr²⁺; while, it was completely inhibited by Ba²⁺ and Co²⁺. The enzyme had a K _m and V _max of 108.0 mg and 79.1 U mL^?1, respectively.     

Purified native haptens of Brucella abortus B19 and B. melitensis 16M reveal the lipopolysaccharide origin of the antigens

Purification of the Brucella polysaccharide referred to as native hapten (NH) and extracted from cells by the autoclaving procedure, was accomplished by ultrafiltration, followed by repetitive gel filtration using high-performance liquid chromatography on a ⪡TSK-G2000-SW⪢ column. The purified NH was analysed by SDS-PAGE, gas-liquid chromatography mass spectroscopy, and ¹³C and ¹H NMR spectroscopy. NH from B. abortus B19 (NH-A) was shown to have a structure identical to that of A polysaccharide from B. abortus 1119-3, a linear homopolymer of α-1,2-linked-4,6-dideoxy-4-formamido-D-mannopyrannosyl residues.The structure of the NH from B. melitensis 16M (NH-M) was identified as a linear homopolysaccharide of the same sugar but composed of a pentasaccharide repeating unit in which four α1,2-linked-4,6-dideoxy-4-formamido-D-mannopyrannosyl residues are linked α-1,3 to the last monosaccharide of the sequence. This structure is similar to that determined for the Brucella M polysaccharide from B. melitensis 16M. The discovery in highly purified NH preparations of covalently bound monosaccharides characteristics of lipopolysaccharide inner core regions e.g., quinovosamine, mannose and 3-deoxy-D-manno-octulosonate (KDO), indicates that this polysaccharide is derived from lipopolysaccharides (LPS) by hydrolytic conditions fortuitously generated during the extraction protocol.The antigenically important polysaccharides of Brucelle are now established to be either A or M antigens. Polysaccharide B is a cyclic glucan with no structural or serological relationship were to A for M polysaccharides, its apparent activity in diagnostic tests of infected cattle resuls from O polysaccharide B.     

Influence of Agronomic Practice on Total Phenols,Carotenoids, Chlorophylls Content,and Biological Activities in Dry Herbs Water Macerates

Oregano (Origanum vulgare L.) and thyme (Thymus vulgaris L.) have long been known for their organoleptic properties. Both plants are widely used in cuisine worldwide in fresh and dried form and as a pharmaceutical raw material. The study aimed to assess if the type of cultivation influenced chosen chemical parameters (total polyphenols by Folin-Ciocalteu method; carotenoids and chlorophyll content by Lichtenthaler method), antimicrobial activity (with chosen reference microbial strains) and shaped cytotoxicity (with L929 mouse fibroblasts cell line) in water macerates of dry oregano and thyme. Polyphenols content and antimicrobial activity were higher in water macerates obtained from conventional cultivation (independently from herb species), unlike the pigments in a higher amount in macerates from organic herbs cultivation. Among all tested macerates stronger antimicrobial properties (effective in inhibiting the growth of Pseudomonas aeruginosa, Bacillus cereus and Salmonella enteritidis) and higher cytotoxicity (abilities to diminish the growth of L929 fibroblasts cytotoxicity) characterized the conventionally cultivated thyme macerate.     

枸杞功能性成分的提取及其组成的液相色谱分析

对枸杞子中的水溶性多糖成分进行了分离提取及组成分析。采用水提、醇沉等方法提取枸杞子中的多糖成分,用Sevage法除去蛋白质后,用凝胶色谱法进行多糖的纯化,制备3-甲基-1-苯基-2-吡唑啉酮（PMP）衍生物进行液相色谱分析。结果表明,枸杞多糖由鼠李糖、阿拉伯糖、木糖、葡萄糖、半乳糖5种单糖组成,其含量分别为5．28％,17．87％,38．29％,17．82％,20．74％。     

灵芝多糖的结构特征分析

采用沸水回流法从灵芝子实体中提取多糖,经Sevage法除蛋白,乙醇沉淀,离心、透析、膜分离,浓缩、冻干后得灵芝多糖。经HIO4氧化、Smith降解及甲基化反应,并利用多糖及刚果红混合液在碱性溶液中的波长的红移变化,通过UV-VIS,IR,GC,GC-MS,NMR对灵芝水提多糖的结构特征及三螺旋体结构进行分析研究。结果表明:灵芝多糖含有三螺旋体构型,GC-MS分析灵芝多糖的主要单糖组分为葡萄糖,还有少量的半乳糖、甘露糖、木糖和艾杜糖,IR及1HNMR分析多糖为β-构型,HIO4氧化、Smith降解和甲基化分析表明:多糖主要为(1→3)糖苷键连接构型,并伴有少量的1→6位支链键连接的结构,灵芝多糖是由D-葡萄糖单元通过β-(1→3)糖苷键连接葡聚多糖,其主要构型特征为(1→3)β-D-线性连接的骨架结构。     

Effect of Thyme Addition on some Chemical and Biological Properties of Sunflower Oil

Antioxidants are used to prevent oxidative changes and flavor development in oils and fats. The aim of this study is to evaluate the antioxidant effect of adding thyme powder added on sunflower oil during frying at different temperature intervals (250 ± 1 °C). Thyme powders were added to sunflower oil at ratio of 0.5%, 1% and 1.5%, and the frying period were estimated for 2 h at 250 ± 1 °C. The oil samples collected intervals were at 0.5, 1, 1.5, and 2 h and the potatoes were fried in each time. The antioxidant activity of thyme powders was 93.05 %, estimated using DPPH root scanning methods. The values of acid, peroxide and, the saponification, ​​and the fatty acid content were considered criteria for evaluating the effectiveness of thyme powder in improving the quality of sunflower oil during frying. Our results confirmed that the adding thyme powder to sunflower oil improved their chemical properties, leading to decrease the acid, peroxide, and saponification values, and unsaturated fatty acids increased. Examination of serum function of rats fed with fried potatoes in sunflower oil-added thyme powder decreased total cholesterol, low-density lipoprotein, and triglycerides, while high-density lipoprotein increased. Moreover the results confirmed that thyme powder reduces liver and kidney functions compared to the control sample. Therefore, adding thyme to sunflower oil retards oxidative decomposition and improves its quality as a natural antioxidant to prolong oil stability.