Antitumor activity of sulfated extracellular polysaccharides of Ganoderma lucidum from the submerged fermentation broth

Water-soluble extracellular polysaccharides are known to possess weak or no in vitro antitumor activity. In this experiment, a mixture of extracellular Ganoderma lucidum polysaccharides (GLP) from the submerged fermentation broth was sulfated and studied on their antitumor activity. The sulfated GLP performed significant inhibition on the proliferation of assayed carcinoma cells in a dose-dependent manner, and present a degree of substitution-dependent suppressing to HepG2 cells. Meanwhile, the sulfated GLP presented remarkable but not dose-dependent inhibition on Heps hepatona in mice. With same degree of substitution, the sulfation protocol with aminosulfonic acid-pyridine yielded GLP sulfates with higher activity on HepG2 cells. In comparison, the native GLP showed no or little antitumor activity on the assayed cell lines but remarkable inhibition on suppressing the proliferation of rat Heps. The highest in vivo inhibition rate of 55.5% provided by sulfated GLP was observed on suppressing the proliferation of rat Heps.     

灵芝蛋白类活性成分的研究进展

灵芝蛋白质是除了灵芝多糖和三萜外,另一类重要的活性物质。它不仅是灵芝生命活动的重要物质而且具有独特免疫原性使其在免疫调节和抗肿瘤病等方面具显著效果。近年来,国内外在灵芝蛋白质的结构特征和生物活性等研究方面已取得显著的成效。本文综述了灵芝蛋白类活性物质的种类和特征,叙述了相关的药理功能,探讨了灵芝蛋白质研究的前景和意义。     

冬虫夏草与韩芝液体发酵混合培养特性研究

平板培养基上比较了冬虫夏草、韩芝及树舌的菌丝生长速度和对淀粉的利用情况,结果显示三者的生长速度分别达到1.28、0.64和0.55 cm/d,且韩芝与树舌对淀粉的利用均优于冬虫夏草;混合培养结果得出菌种的最佳菌龄比为虫草:韩芝=3 d:5 d,最佳的接种比例为2:3;正交试验混合发酵最佳的培养基配方(%):葡萄糖1.0、淀粉2.0、黄豆粉2.0、酵母粉0.5,在此条件下,菌丝生物量达到最大值1.97 g/100 mL。     

紫芝超细粉挥发油成分GC-MS分析

将紫芝Ganoderma sinense(“闽紫96”,HMAS 77207,阔叶树枝桠材或边材栽培)超微粉碎加工成500目的子实体超细粉。激光粒度分析表明:紫芝超细粉的表面积到834．18m~2/kg,中位径(D50)为12．68μm。气质联用分析结果表明:从紫芝超细粉挥发油中分离到35种成分,出峰时间为7．872～30．239min,确定了其中32个成分,占相对含量的88．642％;绝大多数成分为脂肪族化合物(67．216％),主要成分(20．67％)为2,6-二叔丁基一苯酚为芳香族化合物,还检出2种杂环族化合物。     

灵芝连作土壤真菌群落分析

本研究通过Illumina MiSeq平台对邻近野生土壤（GL0）、种植1年（GL1）、2年（GL2）和4年（GL4）的灵芝覆土进行ITS1扩增子测序,分析灵芝连作覆土中真菌群落的变化,为揭示灵芝连作障碍的机理提供理论依据。研究结果表明,12个灵芝土壤样本一共获得349 642条有效序列,经聚类得到2 426个OTU,分别隶属于8门、28纲、64目、127科、233属和449种。在门水平上,随着连作年限的增加,灵芝覆土中真菌的多样性水平逐渐减低,在GL4组中只含有担子菌门（Basidiomycota,占85.03%）、子囊菌门（Ascomycota,占14.77%）和少量被孢霉门（Mortierellomycota,占0.20%）。其中,担子菌门的相对丰度随着连作年限的增加显著增加,子囊菌门的相对丰度显著减少,而被孢霉门的相对丰度无显著性差异。在属水平上,担子菌门灵芝属Ganoderma的相对丰度随着连作年限的增加而极显著增加,子囊菌门仅青霉属Penicillium菌群的相对丰度呈现先减少后增加的趋势,在GL4组中相对于GL0组其相对丰度增加了57.92%,表明青霉属可能是引发该地区灵芝连作障碍的重要菌群,该研究为探索灵芝连作障碍的分子机制和生态防控提供了理论依据。     

海南习用药材无柄灵芝质量标准研究

为建立海南习用药材无柄灵芝的质量标准,依据《中国药典》2015年版中药质量标准研制的要求,对无柄灵芝药材的性状、鉴别、检查、浸出物、灰分、含量测定等项目进行研究。本研究明确了无柄灵芝药材的性状和显微结构特征,建立了专属的薄层鉴别方法;规定无柄灵芝多糖含量不得低于0.40%、三萜与甾醇含量不得低于0.40%、水分不得超过18.0%、总灰分不得超过4.0%、浸出物不得低于2.0%,为无柄灵芝药材质量控制及资源开发利用提供科学依据。     

Structure and chain conformation of five water-soluble derivatives of a β-d-glucan isolated from Ganoderma lucidum

A linear water-insoluble (1→3)-β-d-glucan, coded as GL-IV-I, was isolated from the fruit body of Ganoderma lucidum by extracting with NaOH solution. Its derivatives were prepared by using sulfation, carboxymethylation, hydroxyethylation, hydroxypropylation, and methylation, respectively, and these were labeled as S-GL, CM-GL, HE-GL, HP-GL and M-GL. Five derivatives exhibited good water solubility. Their structures and chain conformations were investigated with infrared spectroscopy, elemental analysis (EA), one- and two-dimensional NMR spectroscopy, laser light scattering (LLS), and size-exclusion chromatography combined with LLS (SEC-LLS). The reactivity of the hydroxyl group of GL-IV-I was ordered as C-6 > C-4 > C-2 for the five derivatives. The degree of substitution (DS) of the derivatives was calculated from EA and NMR spectroscopy to be from 0.32 to 1.18. The weight-average molecular mass (M_w) of GL-IV-I, S-GL, CM-GL, HE-GL, HP-GL, and M-GL was 13.3 × 10⁴, 10.1 × 10⁴, 6.3 × 10⁴, 7.2 × 10⁴, 5.1 × 10⁴, and 14.1 × 10⁴, respectively. The conformation analysis studies revealed that GL-IV-I exists as a compact coil in dimethyl sulfoxide, whereas the five derivatives are slightly expanded flexible chains in 0.9% aqueous NaCl solution.     

Cytotoxic activity induced by crude extracts of Ganoderma lucidum (W. Curt.: Fr.) P. Karst. on mouse myeloma cancer cell-line

Ganoderma lucidum powder using hot water and methanol extraction methods indicated a twofold more active cytotoxic activity with IC₅₀ of 44 ± 3.8 μg/ml in the latter method. The representative dose-response curves of the G. lucidum crude extracts on J558 cell-lines revealed that there were great similarities between the curves which reflected rapid killing activities. The percentage viability of the J558 cell exposed to these crude extracts was dose dependent only up to 150 μg/ml. After which, there was no significant reduction when the dose was increased to 200 or 400 μg/ml. The morphological alterations induced by the crude extract were examined under the phase contrast, fluorescent and electron microscopy. When J558 cells were treated with doses higher than 50 μg/ml of the crude extract, obvious morphological changes and apoptosis occurred after 72 h. At 400 μg/ml, most of the cells showed necrosis characterized as small fragments with uniformly stained red nuclei. The apoptotic and necrotic cells increased by 16.5 and 29.1%, respectively whereas the viable cells decreased by as much as 45.6. The mode of cell death via apoptosis was 3.6% higher than necrosis. However, these morphological changes were not observed in the case of 3T3 cells. Results obtained from scanning electron microscopy and transmission electron microscopy further confirmed the occurrence of various apoptotic and necrotic features.     

Growth Characteristics of Polyporales Mushrooms for the Mycelial Mat Formation

Mushroom strains of Polyporales from the genera Coriolus, Trametes, Pycnoporus, Ganoderma, and Formitella were explored in terms of mycelial growth characteristics for the application of mushroom mycelia as alternative sources of materials replacing fossil fuel-based materials. Among the 64 strains of Polyporales, G. lucidum LBS5496GL was selected as the best candidate because it showed fast mycelial growth with high mycelial strength in both the sawdust-based solid medium and the potato dextrose liquid plate medium. Some of the Polyporales in this study have shown good mycelial growth, however, they mostly formed mycelial mat of weak physical strength. The higher physical strength of mycelial mat by G. lucidum LBS5496GL was attributed to its thick hyphae with the diameter of 13 µm as revealed by scanning electron microscopic analysis whereas the hyphae of others exhibited less than 2 µm. Glycerol and skim milk supported the best mycelial growth of LBS5496GL as a carbon and a nitrogen source, respectively.     

Anti-hepatitis B activities of ganoderic acid from Ganoderma lucidum

Ganoderic acid, from Ganoderma lucidum, at 8 μg/ml inhibited replication of hepatitis B virus (HBV) in HepG2215 cells over 8 days. Production of HBV surface antigen and HBV e antigen were 20 and 44% of controls without ganoderic acid. Male KM mice were significantly protected from liver injury, induced with carbon tetrachloride, by treatment with ganoderic acid at 10 mg and 30 mg/kg·d (by intravenous injection) 7 days. Ganoderic acid at the same dosage also significantly protected the mice from liver injury induced by M. bovis BCG plus lipopolysaccharide (from Escherichia coli 0127:B8).