较高温度对灵芝培养特性及灵芝酸产量的影响

灵芝具有重要的药用价值,培养条件的优化可提高灵芝中各种活性成分的含量。探讨了较高温度对灵芝发酵过程中的生理指标及灵芝酸产量的影响。结果表明,较高的发酵温度虽然降低了灵芝生物量,但提高了淀粉酶﹑漆酶﹑纤维素酶的酶活。发酵第4天,36℃培养条件下淀粉酶酶活达到最大值,比30℃发酵条件下的酶活提高了22.61%。发酵第10天,34℃培养条件下的漆酶酶活﹑纤维素酶酶活﹑可溶性糖含量和灵芝酸产量都达到最大值,相对于30℃的培养条件,分别提高了51.12%、30.41%、13.21%和45.23%。结果表明较高温度发酵有利于酶活和灵芝酸含量的提高,该研究为灵芝的高温发酵提供了新的思路和理论指导。     

灵芝胞外多糖分批发酵动力学模型

利用分批发酵研究了灵芝（Ganoderma lucidum）胞外多糖的合成特性,结果表明Ganoderma lucidum多糖合成和菌体生长呈部分生长关联型。菌体干重、胞外多糖分别达到15.56g·L^-1<、3.02g·L^-1<,胞外多糖对细胞干重得率系数(Y_p/x）为0.19。根据分批发酵试验结果采用Logistic方程、Luedeking-Piret方程和类似Luedeking-Piret方程,得到了描述灵芝生长、胞外多糖以及葡萄糖底物消耗分批发酵动力学模型。同时在初始葡萄糖变化较大范围内,试验数据与模型预测值进行了比较拟合,平均相对误差小于5％,表现出很好的适用性。表明该动力学模型对指导灵芝胞外多糖的发酵生产具有实际意义。     

过表达腺苷生物合成正相关基因GlPNP提高灵芝腺苷的含量

【背景】灵芝被纳入我国“药食同源”试点名单,腺苷作为其主要活性物质之一,在免疫调节、抗炎、抗癌等方面发挥着重要作用。【目的】调控腺苷生物合成关键酶基因的表达来提高灵芝腺苷产量。【方法】将不同培养时间阶段腺苷合成酶基因(包括5-氨基咪唑-4-甲酰胺核糖核苷酸甲酰转移酶GlATIC、嘌呤核苷磷酸化酶GlPNP、腺苷激酶GlADK)的表达量与腺苷含量相关联,筛选出与灵芝腺苷含量呈正相关的关键酶基因。克隆关键酶基因并在灵芝中过表达,探究关键酶基因过表达对灵芝腺苷积累的影响。【结果】GlPNP的表达与灵芝腺苷含量呈正相关。GlPNP的cDNA全长为969 bp,预测GlPNP蛋白的相对分子量为34.6 kDa,呈三聚体的四元结构。研究结果表明,过表达菌株中GlPNP的表达量在第4天比野生型菌株(WT)上调了2.9-3.9倍,与含空载体的菌株(CK)相比,腺苷含量分别提高了78%和63%。【结论】过表达嘌呤核苷磷酸化酶是提高灵芝腺苷产量的一种有效手段。     

影响灵芝三萜类物质液态发酵合成的关键因素研究进展

灵芝是我国名贵的食药两用型菌类,具有广泛的药用价值,其三萜类物质为灵芝中最主要的药理活性物质之一。灵芝液态发酵因具有生长周期短、环境条件可控、目标产物质量稳定及易实现规模化制备等特点而成为获得灵芝三萜类物质最有前景的方式。灵芝三萜代谢途径、发酵工艺及参数、溶解氧控制等是影响灵芝三萜类物质液态发酵合成的关键因素。本文总结了灵芝三萜生物合成的代谢途径和相关的酶(基因)、液态发酵方式和发酵参数调节的溶解氧控制这3个层面对灵芝三萜类物质生物合成的影响,并对今后的研究方向进行了展望,为液态培养灵芝三萜类物质调控及高产提供参考,也为下一步研究提供借鉴。     

A Study on the Synthetic Characteristics of the Extracellular Polysaccharide (EPS) of Ganoderma lucidum Cultured in Batch Fermentation Using a Kinetic Model

The synthetic characteristics of the extracellular polysaccharide (EPS) of Ganoderma lucidum in batch fermentation were studied. The result showed that the production of EPS was partially growth-associated. The cell dry weight (CDW) and EPS reached 15.56 g·L⁻¹ and 3.02 g·L⁻¹, respectively. The yield of EPS to cell dry weight (Y_p/x) was 0.19. On the basis of the test results of batch fermentation, a kinetic model was proposed by using the Logistic equation for cell growth, the Luedeking–Piret equation for EPS production, and the Luedeking-piret-like equation for the consumption of glucose as substrate. The calculated results using these models were satisfactorily compared with the experimental data under various concentrations of glucose, and the average of relative errors was found to be not more than 5%. The kinetic model had practical guidance interesting in producing PES by Ganoderma lucidum.     

海南灵芝化学成分研究

为探讨海南灵芝(Ganoderma hainanense Zhao,Xu et Zhang)的化学成分,采用柱层析技术从海南灵芝乙醇提取物中分离得到14个化合物。经波谱分析鉴定其结构分别为:巴西红厚壳素(1)、6-脱氧巴西红厚壳素(2)、7,8-二甲基咯嗪(3)、5,8-过氧麦角固醇(4)、3β,5α,9α-三羟基麦角甾-7,22-二烯-6-酮(5)、麦角甾-7,22-二烯-3β-醇(6)、麦角甾-7,22-二烯-3-酮(7)、7α-甲氧基-5α,6α-环氧麦角甾-8(14),22-二烯-3β-醇(8)、麦角甾-7,22-二烯-3β,5α-6β-三醇(9)、3-吲哚甲酸(10)、3,4-二羟基苯甲酸(11)、对羟基苯甲酸(12)、3,4-二羟基苯甲酸甲酯(13)和正二十六烷酸(14)。所有化合物均为首次从海南灵芝中分离得到。     

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).     

Significance of inoculation density control in production of polysaccharide and ganoderic acid by submerged culture of Ganoderma lucidum

Control of inoculation density was significant for cell growth, morphology, and production of polysaccharide and ganoderic acid in submerged culture of the higher fungus Ganoderma lucidum. A maximal cell concentration of 15.7 g dry cell weight (DW)/l was obtained at an inoculation density of 330 mg DW/l. For inoculation density within the range of 70–670 mg DW/l, a large inoculation density led to a small pellet size and high production of extracellular and intracellular polysaccharides, while a relatively big pellet size and high accumulation of ganoderic acid were observed at a low inoculation density. It was also shown that small pellet size resulted in high polysaccharide production, while large pellet size led to high production of ganoderic acid.     

Cytotoxic triterpenoids from Ganoderma lucidum

A systematic study of the metabolites in Ganoderma lucidum led to isolation of 43 triterpenoids, six of them (1-6) are hitherto unknown. The structures of the latter were elucidated on the basis of spectroscopic studies and comparison with the known related compounds. All of the compounds were assayed for their inhibitory activities against human HeLa cervical cancer cell lines. Some compounds exhibit significant cytotoxicity, and their structure-activity relationships are discussed.     

A new ecdysteroid and other constituents from two Dioscorea species

Phytochemical investigation of the rhizome of Dioscorea dumetorum has led to the isolation by several chromatographic steps on normal and reversed phase silica gel of a new ecdysteroid, (20R)-5β,11α,20-trihydroxyecdysone (1), and two known ecdysteroids, ajugasterone C (2) and herkesterone (3). Their structures were determined by spectroscopic methods including 1D- and 2D-NMR (COSY, TOCSY, HSQC and HMBC). This is the first report on the occurrence of phytoecdysteroids in the Dioscoreaceae family. These compounds were devoid of antifungal activity against three Candida species (Candida albicans, Candida glabrata and Candida tropicalis, MIC > 200 μg/ml).     

灵芝UDP-葡萄糖4-差向异构酶酶学性质研究及其应用

【背景】灵芝多糖是灵芝的重要活性物质之一。UDP-葡萄糖4-差向异构酶（UDP-glucose 4-epimerase,UGE,EC 5.1.3.2）是灵芝多糖合成途径中糖供体生成的重要酶,其参与了UDP-葡萄糖与UDP-半乳糖的相互转化,与多糖中半乳糖残基含量密切相关。【目的】通过对来源于灵芝的UGE基因进行异源表达,丰富灵芝多糖糖供体合成途径重要酶的酶学特性信息,深入了解灵芝多糖代谢合成途径。【方法】以灵芝菌株（Ganoderma lingzhi） CGMCC 5.26的cDNA为模板,克隆得到UGE基因GL30389,并在Escherichia coli BL21（DE3）中诱导表达,产物纯化后进行酶学性质、酶动力学、底物专一性及转化率的研究。【结果】灵芝UGE的分子量为45 kDa。最适反应pH值为6.0,在pH 7.0—9.0范围内有较好的稳定性;最适反应温度为30℃,温度在40℃时稳定性最好。Fe²⁺和Mg²⁺对UGE有激活作用。以UDP-葡萄糖为底物时,K_m为0.824 mmol/L,V_max为769.230 μmol/（L·min）,k_cat为1.333 s^—1,k_cat/K_m为1.618 L/（mmol·s）。灵芝UGE对D-葡萄糖、半乳糖醛酸及N-乙酰葡萄糖胺有催化活性。通过优化pH、温度、底物与酶的配比、添加金属离子将转化率从16.0%提升至39.4%。【结论】灵芝UGE与植物来源的UGE酶学性质较为相似,其催化效率优于大部分细菌来源的UGE。本研究丰富了灵芝多糖糖供体合成途径重要酶的酶学特性信息,有利于深入了解灵芝多糖代谢合成途径。     

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.     

荔枝草中一个新的倍半萜

为了研究荔枝草全草中的倍半萜类化学成分,该文运用大孔吸附树脂D101、MCI、硅胶和反相HPLC等色谱技术对其进行分离纯化,根据HR-ESI-MS、NMR波谱数据鉴定了化合物的结构。结果表明:从荔枝草全草95%乙醇提取物的乙酸乙酯部位中分离得到3个桉叶烷型倍半萜类化合物,分别鉴定为8-ethoxy-eudebeiolide B(1)、salplebeone D(2)、salplebeone A(3)。其中,化合物1为新化合物。     

A new triterpenic acid from the wood rotting fungi

A new triterpenic acid, assigned the trivial name polyporenic acid D, has been isolated from the wood rotting fungus Polyporus officinalis. It has been shown to have the structure 3α-hydroxy-4,4,14α-trimethyl-5α-ergosta-8, 24(28)-dien-26-oic acid.     

灵芝液态发酵胞内外多糖的研究进展

灵芝具有悠久的药用历史,其活性物质灵芝多糖具有广泛的药理活性,利用液态发酵技术获取灵芝多糖展现出独特优势。近年来,关于灵芝多糖液态发酵的研究报道越来越多,引起了研究者的广泛关注。本文从灵芝多糖的生物合成代谢及调控、液态发酵的培养基及工艺优化等方面对近期的研究结果进行总结,以期为灵芝液态发酵制备胞内外多糖及其产业化应用提供参考和依据。     

托光柄菇属一中国新记录种

[背景] 托光柄菇属（Volvopluteus）隶属于光柄菇科（Pluteaceae）,目前世界上仅有4个种。[目的] 调查我国东北地区大型真菌资源。[方法] 采集大型真菌标本,对其形态进行详细的观察和描述,提取DNA,测定rDNA ITS序列,基于最大似然法和贝叶斯法构建系统发育树。[结果] 2019–2020年在吉林省延边朝鲜族自治州敦化市采集的标本中,有6份标本为密执安托光柄菇V.michiganensis,该种真菌此前未在我国发现。在系统发育分析中,采自我国的密执安托光柄菇V.michiganensis与该种的模式标本聚为一个分支。[结论] 密执安托光柄菇V.michiganensis为中国新记录种。     

Domoic acid production by Pseudo-nitzschia australis and Pseudo-nitzschia calliantha isolated from North Chile

The morphology and toxicity of the four ubiquitous species belonging to the genus Pseudo-nitzschia found in mixed blooms of phytoplankton from northern Chilean waters were studied. The phytoplankton samples and cultures obtained were identified by scanning electron microscopy, revealing the presence of Pseudo-nitzschia australis, P. calliantha, P. pseudodelicatissima and P. subfraudulenta. This is the first report of P. calliantha in northern Chile. Toxin analyses using the LC–MS method confirmed the presence of domoic acid in P. australis and P. calliantha. Domoic acid was not detected in cultures of P. subfraudulenta. This study therefore confirms P. australis and P. calliantha as an unequivocal source of domoic acid in Chilean waters. P. australis is probably the most important producer of amnesic shellfish toxin in view of its domoic acid content. However, more research is needed to evaluate the potential for toxin production in P. pseudodelicatissima.     

Fatty acid composition of lipids from mushrooms belonging to the family Boletaceae

The non-polar lipid content and fatty acid (FA) composition of 11 mushroom species of the family Boletaceae were determined. The non-polar lipid content ranged from 2.0 (Leccinum aurantiacum and Boletus erythropus) to 5.4 % (w/w) d.w. (Suillus grevillei) with an average value of 2.9 %. More than 25 different FAs were found in the mushroom lipids. Unsaturated FAs, mainly linoleic and oleic acids, accounted for about 83 % of the total FAs, while palmitic acid was the main saturated FA. Some FAs are identified for the first time in Boletaceae and in higher Basidiomycetes (cis-11,12-methyleneoctadecanoic acid, 7-cis,10-cis hexadecadienoic) or in fungi (cis-11,12-methyleneoctadecanoic acid). There were significant differences (P < 0.05) in the contents of specific FAs between mushroom species.