An antitumor pectic polysaccharide from Feronia limonia

An acidic heteropolysaccharide has been isolated from the tropical angiosperm Feronia limonia syn. F. elephantum (family: Rutaceae). A partially carboxymethylated alpha-(1-4) polygalacturonan backbone structure with 2- and 2,4-O-alpha-L-rhamnopyranosyl, 2- and 2, 3-O-alpha-L-arabinofuranosyl and 3-, 2,4-and terminal alpha-D-galactopyranosyl bearing side chains has been tentatively assigned. The preliminary study in the murine model showed some significant in vivo Ehrlich ascites carcinoma cell growth inhibition.     

Comparison of antitumor activities of different polysaccharide fractions from the stems of Dendrobium nobile Lindl

The antitumor activities of extracted polysaccharide fractions from the stems of Dendrobium nobile Lindl were investigated. Polysaccharides were sequentially extracted from the stems of D. nobile to obtain three fractions, i.e. water extract fraction (DNP-W), 5% NaOH extract fraction (DNP-OH) and 5% HCl extract fraction (DNP-H). Further the DNP-W was isolated to give six sub-fractions (DNP-W1, DNP-W2, DNP-W3, DNP-W4, DNP-W5 and DNP-W6) by anion-exchange chromatography. The monosaccharide profile, protein content, uronic acid content, total carbohydrate content, viscosity and molecular weight of nine polysaccharide fractions were analyzed. Both the in vivo and in vitro antitumor activities of nine polysaccharide fractions were evaluated and compared. Results indicated that DNP-W1 and DNP-W3 exhibited high antitumor activities against Sarcoma 180 in vivo and HL-60 in vitro. The results suggested that DNP-W1 and DNP-W3 could be considered as an effective natural antitumor source.     

Structural analysis of a pectic polysaccharide from the leaves of Diospyros kaki

A pectic polysaccharide DL-2A with a molar mass of 8.5 x 10(5), was obtained from the boiling water extract of Diospyros kaki leaves. It had [alpha]20D -21.8 degrees (c 0.22, H2O) and consisted of rhamnose, arabinose, galactose, xylose and galacturonic acid units in the molar ratio of 0.4:3.4:2.4:1.0:0.8, along with traces of glucuronic acid. About 16.7% of galacturonic acid existed as the methyl ester. A combination of linkage analyses, periodate oxidation, partial acid hydrolysis, selective lithium-degraded reaction, ESIMS, 1H- and 13C- NMR spectral analyses revealed its structural features. It was found that DL-2A possessed an alpha-(1-->4)-galacturonan backbone with some insertions of alpha-1,2-Rhap residues. The side-chains of arabino-3,6-galactan were attached to the backbone via O-4 of Rhap residues and O-3 of GalAp residues, while 4-linked xylose residues (forming short linear chains) were directly linked to O-4 of rhamnose residues, not as part of the xylogalacturonan. These novel structural features enlarge the knowledge on the fine structure of pectic substances in the plant kingdom.     

A polysaccharide from Lichina pygmaea and L. confinis supports the recognition of Lichinomycetes

The lichen-forming order Lichinales, generally characterized by prototunicate asci and the development of thalli with cyanobacteria, has recently been recognized as a separate class of ascomycetes, Lichinomycetes, as a result of molecular phylogenetic studies. As alkali and water-soluble (F1SS) polysaccharides reflect phylogeny in other ascomycetes, a polysaccharide from Lichina pygmaea and L. confinis was purified and characterized to investigate whether these F1SS compounds in the Lichinomycetes were distinctive. Nuclear magnetic resonance (NMR) spectroscopy and chemical analyses revealed this as a galactomannan comprising a repeating unit consisting of an α-(1→6)-mannan backbone, mainly substituted by single α-galactofuranose residues at the O-2- or the O-2,4- positions linked to a small mannan core. With the exception of the trisubstituted mannopyranose residues previously described in polysaccharides from other lichens belonging to orders now placed in Lecanoromycetes, the structure of this galactomannan most closely resembles those found in several members of the Onygenales in Eurotiomycetes. Our polysaccharide data support molecular studies showing that Lichina species are remote from Lecanoromycetes as the galactofuranose residues are in the α-configuration. That the Lichinomycetes were part of an ancestral lichenized group can not be established from the present data because the extracted polysaccharide does not have the galactofuranose residue in the β configuration; however, the data does suggest that an ancestor of the Lichinomycetes contained a mannan and was part of an early radiation in the ascomycetes.     

l-Galactose replaces l-fucose in the pectic polysaccharide rhamnogalacturonan II synthesized by the l-fucose-deficient mur1 Arabidopsis mutant

Arabidopsis thaliana mur1 is a dwarf mutant with altered cell-wall properties, in which l-fucose is partially replaced by l-galactose in the xyloglucan and glycoproteins. We found that the mur1 mutation also affects the primary structure of the pectic polysaccharide rhamnogalacturonan II (RG-II). In mur1 RG-II a non-reducing terminal 2-O-methyl l-galactosyl residue and a 3,4-linked l-galactosyl residue replace the non-reducing terminal 2-O-methyl l-fucosyl residue and the 3,4-linked l-fucosyl residue, respectively, that are present in wild-type RG-II. Furthermore, we found that a terminal non-reducing l-galactosyl residue, rather than the previously reported d-galactosyl residue, is present on the 2-O-methyl xylose-containing side chain of RG-II in both wild type and mur1 plants. Approximately 95% of the RG-II from wild type and mur1 plants is solubilized as a high-molecular-weight (>100 kDa) complex, by treating walls with aqueous potassium phosphate. The molecular mass of RG-II in this complex was reduced to 5–10 kDa by treatment with endopolygalacturonase, providing additional evidence that RG-II is covalently linked to homogalacturonan. The results of this study provide additional information on the structure of RG-II and the role of this pectic polysaccharide in the plant cell wall.Abbreviations AIR Alcohol-insoluble residue - d-Gal d-Galactosyl - EPG Endopolygalacturonase - ESI–MS Electrospray ionization mass spectrometry - GC–MS Gas chromatography–mass spectrometry - ¹H-NMR Proton nuclear magnetic resonance spectroscopy - l-Fuc l-Fucosyl - l-Gal l-Galactosyl - 2-O-MeFuc 2-O-Methyl l-fucosyl - 2-O-MeGal 2-O-Methyl l-galactosyl - 2-O-MeXyl 2-O-Methyl d-xylosyl - MWCO Molecular weight cut-off - RG-II Rhamnogalacturonan II - ppm Parts per million - RI Refractive index - SEC Size-exclusion chromatography - TFA Trifluoroacetic acid - WT Wild type     

Antifungal activity of tea tree oil from Melaleuca alternifolia against Trichophyton equinum: An in vivo assay

Dermatophytes are a group of keratinophilic and keratinolytic molds, some of which are responsible for ringworm. Among them Trichophyton equinum, which mostly infects equids, can cause extensive outbreaks in stud farms. The conventional treatment of equine trichophytosis is topic, based upon medicated shampoos to reduce the spread of infection among the animals. Nevertheless the popularity of phytotherapy is at an all-time peak, and the interest for natural alternatives or complements to conventional drug therapy is challenging both in human and veterinary field. Among herbal remedia Tea Tree Oil (TTO) shows a wide range of antimicrobial activities. A randomized open clinical trial was carried out on 60 thoroughbred breeding horses affected by equine ringworm. The animals were randomly divided into 2 groups of 30 subjects. Diagnostic criteria were the presence of clinical signs and positive T. equinum culture. Specificity control using TTO mixture in 5 not dermatophyte affected animals was achieved also. The antimycotic activity against T. equinum of a mixture containing 25% TTO in sweet almond oil, was evaluated in vivo treating 30 subjects, the others were administered enilconazole 2% solution. The animals of both groups were topically treated twice a day for 15 days with a 25% mixture of TTO diluted in sweet almond oil and every 3 days, four times with enilconazole rinses, respectively. The clinical and mycological outcome were evaluated at day 30 from the start of the treatments. Data analysis was performed by chi square test. All the treated animals showed complete clinical and aetiological healing. Part of control subjects also, showed an improvement and none of them exacerbate the lesions. This therapeutic protocol appears to be effective and versatile, being applicable immediately after physical examination, prior to have the laboratory response. It could be an alternative for practitioners interested in herbal medicines, contributing to fulfill the gap existing between in vitro and clinical studies.     

三孢布拉霉crgA启动子的克隆和活性分析

【背景】CrgA是三孢布拉霉(Blakesleatrispora,Bt)中调控类胡萝卜素合成的关键负调控因子,其表达水平会影响类胡萝卜素的合成。【目的】克隆三孢布拉霉crgA启动子并分析其活性,为进一步解析CrgA表达调控机制奠定基础。【方法】通过综合微生物基因组(integrated microbial genomes, IMG)数据库提供的基因组序列,克隆crgA翻译起始位点上游2 000 bp序列,分析其顺式调控元件和转录起始区域预测,通过RT-qPCR分析不同光照时间对三孢布拉霉crgA相对转录水平的影响;构建4个不同长度的crgA启动子截短序列驱动的GUS-mGFP5重组表达载体p1303-procrgAF、F1、F2和F3,利用农杆菌侵染整合到三孢布拉霉基因组中,在黑暗和光照条件下测定β-D-葡萄糖苷酸酶(β-D-glucuronidase,GUS)酶活性并观察荧光信号。【结果】crgA启动子不仅包含基础的TATA-box和CAAT-box元件,还包括多个与光响应相关的元件。观察荧光结果显示CaMV35S和构建的4个突变启动子均能在三孢布拉霉体内驱动下游基因表达,检测GUS...     

Water-soluble polysaccharide from Bupleurum chinense DC: Isolation, structural features and antioxidant activity

The water-soluble polysaccharide (BCPS-1) was isolated from Bupleurum chinense DC. BCPS-1 (M_w = 29 kDa) was composed of Ara; Gal; Glc with a molar ratio of 2.1:2.5:1. According to FT-IR, partial acid hydrolysis, periodate oxidation and Smith degradation, methylation and GC-MS analysis, the results indicate BCPS-1 had a backbone of (1→5)-linked Ara, (1→4)-linked Gal and (1→3)-linked Gal residues with occasionally branches at O-6. The branches were composed of (1→4)-linked Glc, and terminated with Gal residues. The in vitro antioxidant activity evaluated by DPPH radical scavenging method showed that BCPS-1 had a significant antioxidant effect in a concentration-dependent manner.     

An unusual Xanthophyllomyces strain from leaves of Eucalyptus globulus in Chile

Xanthophyllomyces sp. was isolated as an epiphytic red yeast from leaves of Eucalyptus glo-bulus in Concepción, Chile. Sexual reproduction was by basidiospores produced from one or rarely two metabasidia arising from a yeast cell without preceding paedogamy. The main carotenoid pigment was astaxanthin. This isolate did not cluster with the X. dendrorhous complex (including Phaffia rhodozyma) in ITS and 26S rDNA-based phylogenetic analyses. The phylloplane may be a further habitat for Xanthophyllomyces, in addition to the well-known spring sap-flows of deciduous trees and the recently-characterised ascostromata of Cyttaria hariotii.     

鹰爪花中生物碱类成分研究

为了解鹰爪花(Artabotrys hexapetalus)的化学成分,采用各种柱层析和色谱方法从鹰爪花茎中分离鉴定了12种生物碱类化合物,分别为:光千金藤定碱(1)、四氢非洲防己碱(2)、四氢药根碱(3)、前莲碱(4)、奥可梯木种碱(5)、华防己碱(6)、深山黄堇碱(7)、光千金藤碱(8)、鹰爪花碱(9)、lanuginolide(10)、isoscoulerine(11)和N-甲基阿西米洛宾(12)。其中化合物1~7均为首次从鹰爪花属植物中分离得到。化合物10对白色葡萄球菌(Staphylococcus albus)和蜡状芽孢杆菌(Bacillus cereus)生长都具有较好的抑制作用,MIC值分别为20和10μg m L–1。     

一株产虫草素的冬虫夏草内生真菌 Bionectria ochroleuca OS17

虫草素是一种核苷类似物,具有多种药理作用。利用虫草素产生菌发酵的方式获得虫草素,可能是产生虫草素的一种新方法。为筛选能产虫草素的内生真菌,从中国传统的中药冬虫夏草子实体中共分离得到42株真菌,经PDB液体培养后,采用高效液相色谱（HPLC）鉴定其代谢产物。结果表明：菌株OS17的代谢产物中含有虫草素,且虫草素产量为40.16 mg/L。基于形态学特征和ITS序列分析,菌株OS17被鉴定为淡色生赤壳菌(Bionectria ochroleuca)。这是首次从冬虫夏草中分离得到能产虫草素的生赤壳属,表明了内生真菌具有产生多种生物活性物质的潜力。     

铁皮石斛内生真菌次生代谢产物研究

为了解铁皮石斛(Dendrobium officinale)内生真菌Phyllosticta aristolochiicola的次生代谢产物,从该真菌中分离得到15个化合物,经波谱分析分别鉴定为N-methyl-2-pyrolidinone (1)、环-(甘氨酸-L-脯氨酸)(2)、环-(D-丙氨酸-L-脯氨酸)(3)、环-(L-缬氨酸-L-脯氨酸)(4)、环-(L-亮氨酸-L-脯氨酸)(5)、cyclo-(L-Leu-D-4-hydroxyprolinyl)(6)、环-(L-苯丙氨酸-L-脯氨酸)(7)、环-(L-苯丙氨酸-L-4-羟基脯氨酸)(8)、环-(L-酪氨酸-L-脯氨酸)(9)、环-(L-苯丙氨酸-L-亮氨酸)(10)、啤酒甾醇(11)、对羟基苯乙醇(12)、对羟基苯乙酸(13)、(2S,3R)-1-(4-羟基苯基)丁烷-2,3-二醇(14)和(2R,3S)-1-苯基丁烷-2,3-二醇(15)。采用MTS法检测抗肿瘤活性表明,化合物2、10和14对HL-60、A-549、SMMC-7721、MCF-7和SW-480细胞株具有一定的抑制活性。     

毛竹PeSCL3基因表达特征及其启动子活性研究

为探讨毛竹(Phyllostachys edulis)SCL3基因的表达特征及其启动子活性,采用同源克隆的方法从毛竹中分离到SCL3同源基因Pe SCL3的编码区(ORF)和上游启动子序列(Pe SCL3p)。序列分析表明,Pe SCL3的ORF为1335 bp,推测编码含444氨基酸的蛋白,该蛋白与水稻(Oryza sativa)的SCL3同源性高达93.9%。Pe SCL3p长度为1358 bp,含有脱落酸(ABA)应答元件ABRE、赤霉素(GA3)应答元件GARE-motif和P-box、干旱诱导MYB结合位点等多种作用元件。实时定量PCR分析结果表明,Pe SCL3在毛竹叶中的表达丰度最高,其次是茎和根,而鞘中的最低;Pe SCL3的表达受GA3的抑制,受ABA、Na Cl和干旱的诱导。转Pe SCL3p∷GUS拟南芥(Arabidposis thaliana)的GUS染色结果表明,根尖、顶端生长点和子叶叶柄均被染成蓝色,尤其根尖的染色最深。这表明Pe SCL3对毛竹的生长发育,尤其是根系,可能起着重要的调控作用。     

Isariotins G–J from cultures of the Lepidoptera pathogenic fungus Isaria tenuipes

Four new alkaloids isariotins G–J (1–4), together with the known isariotins, were isolated from cultures of the Lepidoptera pathogenic fungus Isaria tenuipes. The structures were elucidated on the basis of NMR spectroscopic and mass spectrometry data. Compounds 1–4 exhibited antimalarial and cytotoxic activities.     

Structural studies of the O-antigen polysaccharide from the enteroinvasive Escherichia coli O173

The structure of the O-antigen polysaccharide (PS) from Escherichia coli O173 has been investigated. Sugar and methylation analyses, electrospray ionisation mass spectrometry together with ¹H, ³¹P and ¹³C NMR spectroscopy were the main methods used. The structure of the pentasaccharide repeating unit of the PS was found to be:

By treatment with 48% HF the phosphoric diester linkage was cleaved together with the glycosidic linkage of the fucosyl group, rendering a tetrasaccharide with the structure:

     

New species of Phaeoramularia, Pseudocercospora and Stenella from Western Ghats of India

Four new species of the hyphomycete genera Phaeoramularia viz. Ph. caesalpinae, Pseudocercospora viz., Ps. tiliacearum, Stenella, viz. S. argyreiae and S. grewiae occurring on Caesalpinia bonducella Fleming (Caesalpiniaceae), Grewia sp. (Tiliaceae), Argyrea sp. Lour (Convolvulaceae) and Grewia sp. L. (Tiliaceae), respectively are described and illustrated here. All these fungi were collected from Western Ghats of India.     

宁前胡内生真菌Fusarium tricinctum固体发酵代谢产物研究

为了解宁前胡内生真菌Fusarium tricinctum固体发酵代谢产物的化学成分,采用硅胶色谱、中压以及制备液相等方法对其进行分离鉴定和抗肿瘤活性研究。结果表明,从其中分离得到14个化合物,结构鉴定分别为fungerin (1)、fusagerins F (2)、6-[(3R)-3-hydroxybutan-2-yl]-5-(hydroxymethyl)-4-methoxy-2H-pyran-2-one (3)、6-[(3S)-3-hydroxybutan-2-yl]-5-(hydroxymethyl)-4-methoxy-2H-pyran-2-one(4)、6α-chlamydosporol(5)、6β-chlamydosporol(6)、(3R,4R)-(–)-4-hydroxymellein(7)、(3R)-(–)-8-hydroxymellein (8)、enniatin B (9)、enniatin B1 (10)、enniatin A1 (11)、enniatin A (12)、enniatin B3 (13)和enniatin B2 (14)。所有化合物均为首次从宁前胡内生真菌中分离得到。体外抗肿瘤活性试验表明化合物均无显著的细胞毒活性。     

Sulfated polysaccharides from brown seaweeds Saccharina japonica and Undaria pinnatifida: isolation, structural characteristics, and antitumor activity

During the last decade brown seaweeds attracted much attention as a source of polysaccharides, namely laminarans, alginic acids, and sulfated polysaccharides—fucoidans, with various structures and biological activities.In this study, sulfated polysaccharides were isolated from brown seaweeds Saccharina japonica (formerly named Laminaria) and Undaria pinnatifida and their antitumor activity was tested against human breast cancer T-47D and melanoma SK-MEL-28 cell lines.The sulfated polysaccharide form S. japonica was highly branched partially acetylated sulfated galactofucan, built up of (1→3)-α-l-fucose residues. The sulfated polysaccharide from U. pinnatifida was partially acetylated highly sulfated galactofucan consisting of (1→3)- or (1→3);(1→4)-α-l-fucose residues.Fucoidans from S. japonica and U. pinnatifida distinctly inhibited proliferation and colony formation in both breast cancer and melanoma cell lines in a dose-dependent manner. These results indicated that the use of sulfated polysaccharides from brown seaweeds S. japonica and U. pinnatifida might be a potential approach for cancer treatment.