Two perenniporiol derivatives,lanostane-type triterpenoids,from the cultured mycelia of Perenniporia ochroleuca

Two new perenniporiol derivatives, lanostane-type triterpenoids, were isolated from the benzene extract of cultured mycelia of Perenniporia ochroleuca. The structures of the two new compounds were elucidated by spectral data to be 3-O-acetyl-7,11-dihydroperenniporiol and 12β-acetoxyperenniporiol.     

Bioconversion of a ganoderic acid 3-hydroxy-lanosta-8,24-dien-26-oic acid by a crude enzyme from Ganoderma lucidum

Ganoderic acids (GAs) are oxygenated lanostane-type triterpenoids from the traditional medicinal mushroom Ganoderma lucidum and of significant biological activities. Although a ganoderic acid 3-hydroxy-lanosta-8,24-dien-26-oic acid (HLDOA) was found to be biosynthesized from lanosterol, further post-modification of HLDOA is yet unclear. In this work, by using HLDOA as the substrate and a crude enzyme from G. lucidum as the biocatalyst, we observed a new peak in liquid chromatography from the reaction system. The product was purified and identified to be 3-oxo-lanosta-8,24-dien-26-oic acid (OLDOA), which may be converted from HLDOA by a putative dehydrogenase of G. lucidum. The work is useful to future manufacture of GAs as well as their biosynthetic pathway elucidation.     

蜣螂对灵芝发酵菌丝体生长和三萜产物形成的影响

研究了几种药用昆虫对灵芝深层培养过程中细胞生长和三萜产物形成的影响。结果表明,添加不同药用昆虫对灵芝细胞生长和细胞内三萜的形成无显著促进作用(P>0.05),但蜣螂虫粉的添加,能显著促进灵芝胞外三萜的形成(P<0.01)。在添加浓度为5g/L时,胞外三萜的产量从对照的220.2±9.7mg/L提高到304.3±11.8mg/L。结果提示药用昆虫蜣螂虫粉中含有促进灵芝胞外三萜形成的功能因子。     

Improvement of ganoderic acid and Ganoderma polysaccharide biosynthesis by Ganoderma lucidum fermentation under the inducement of Cu2+

The cell growth and total accumulation of bioactive metabolites were significantly improved by Cu²⁺ addition during the submerged fermentation of medicinal mushroom Ganoderma lucidum. A mathematical model, constructed by response surface methodology combination with full factorial design, was applied to study the synergic effect of Cu²⁺ addition concentration and addition time. The optimal Cu²⁺ inducement strategy for the cell growth were different from those for the biosynthesis of ganoderic acid (GA) and Ganoderma polysaccharide. A multiple additions strategy of Cu²⁺ by adding each 1 mM Cu²⁺ on day 2, 6, 8 and 2 mM Cu²⁺ on day 4 was developed to enhance total accumulation of GA and extracellular polysaccharides. The highest GA content reached 3.0 ± 0.1 mg per 100 mg DW, which was increased by 76.5% and 33.9% compared with the control without Cu²⁺ addition and the peak value predicted by the constructed mathematical model, respectively. While, relatively higher addition concentration of Cu²⁺ (i.e., 5 mM) on the culture of day 4 led to higher content and total production of intracellular polysaccharides. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

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).     

Effects of different graphene-based nanomaterials as elicitors on growth and ganoderic acid production by Ganoderma lucidum

Graphene-based nanomaterials (GBNs) have attracted considerable interest nowadays due to their wide range of applications. However, very little attention has been paid to the application of nanomaterials as potential elicitors for production of valuable metabolites. Herein, aiming to earn insight into effects of nanomaterials on secondary metabolite biosynthesis by medicinal fungi, we evaluated the influence of GBNs on growth and production of ganoderic acid (GA) by Ganoderma lucidum in submerged culture. Graphene oxide (GO), reduced graphene oxide (rGO), and rGO/Fe₃O₄ nanocomposite were synthesized successfully and characterized by X-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy analysis. The prepared nanomaterials were added to the culture of G. lucidum at final concentrations of 50, 100, and 150 mg/L on Day 5. The results showed that the elicitation of G. lucidum with GO and rGO decreased the cell dry weight and GA production slightly, especially in higher concentrations. However, rGO/Fe₃O₄ nanocomposite not negatively affected cell growth and improved GA production. G. lucidum growth rate responded to elicitation experiments differently and depended on the type of nanomaterials and their concentrations, but almost all GBNs caused an increase in GA content (mg/100 mg dry weight). Also, field emission scanning electron microscopy morphological study showed that under elicitation, mycelia were more condensed and tightly stacked together. The findings from this study may suggest that GBNs in low concentrations could be applied as elicitors to secondary metabolites production from higher fungus, but further environmental, physiological, and biological studies required.     

l-3-(3-carboxyfuran-4-yl)alanine,a new amino acid from the mushroom Phyllotopsis nidulans

A new amino acid has been discovered in uncombined form in extracts of the fruiting bodies of the mushroom, Phyllotopsis nidulans. Chemical and spectroscopic data support formulation of the structure as l-3-(3-carboxyfuran-4-yl)alanine.     

Karachic acid: A new triterpenoid from Betula utilis

Karachic acid, from the bark of Betula utilis, has been shown to be 3β-hydroxy 6α-acetoxy oleanoic acid, a new triterpenoid of the oleanoic acid type.     

A new epoxidic ganoderic acid and other phytoconstituents from Ganoderma lucidum

A new epoxidic ganoderic acid, 8α,9α-epoxy-3,7,11,15,23-pentaoxo-5α-lanosta-26-oic acid (1), together with the known compounds 3β-hydroxy-7,11,15,23-tetraoxo-5α-lanosta-8-en-26-oic acid (2), ergosta-7,22-diene-3β-yl pentadecanoate (3), ergosta-7,22-diene-3β-ol (4), β-sitosterol (5), fatty acids (6–10), fatty acid ester (11) and octadecane (12) were isolated from the fruiting bodies of Ganoderma lucidum from south India. Their structures were determined by ¹H, ¹³C, ¹³C DEPT, ¹H–¹H COSY, HMBC, HSQC, NOESY NMR, FT-IR, UV–vis and FABMS spectral analysis. Compounds (1–3) exhibited good antifungal activity against Candida albicans in disc diffusion assay (100 μg/disc). Steroid ester (3) showed moderate anti-inflammatory activity (59.7% inhibition, 100 mg/kg body weight) in carrageenan-induced paw edema.     

Secondary metabolites from Ganoderma lucidum and Spongiporus leucomallellus

The hydrodistillates and solvent extracts of the fruit bodies of Ganoderma lucidum (Fr.) P. Karst. and Spongiporus leucomallellus (Murril) A. David were investigated. The constituents in both oils comprised hydrocarbons, monoterpenes, sesquiterpenes, and fatty acids. Major volatiles of G. lucidum were trans-anethol, R-(-)-linalool, S-(+)-carvone and alpha-bisabolol, while the essential oil of S. leucomallellus contained relatively large amounts of R-(-)-1-octene-3-ol, R-(-)-linalool, 1-hepten-3-one and (Z)-nerolidol. From the n-hexane extract of G. lucidum, the steroid ester ergosta-7,22-diene-3beta-yl pentadecanoate could be identified. From S. leucomallellus two constituents showing structures of 3,4-seco-lanostane type triterpene acids were identified as (+)-23-oxo-3,4-seco-lanosta-4(28),7(8),9(11),24(31)-tetraene-3,26-dicarboxylic acid and (+)-20-hydroxy-23-oxo-3,4-seco-lanosta-4(28),7(8),9(11),24(31)-tetraene3,26-dicarboxylic acid, respectively. Cytotoxicity and antimicrobial activity of selected compounds were investigated using standard tests.     

Efficient biosynthesis of antitumor ganoderic acid HLDOA using a dual tunable system for optimizing the expression of CYP5150L8 and a Ganoderma P450 reductase

Ganoderic acid 3-hydroxy-lanosta-8,24-dien-26-oic acid (GA-HLDOA), an antitumor triterpenoid from the traditional Chinese medicinal higher fungus Ganoderma lucidum, is considered as a key precursor for biosynthesizing other ganoderic acids (GAs) with superior antitumor activities. Our previous study identified CYP5150L8 from G. lucidum as a lanosterol oxidase, and achieved heterologous biosynthesis of GA-HLDOA in Saccharomyces cerevisiae. However, low production of GA-HLDOA in either G. lucidum or heterologous host hindered its further investigation and application. In this study, we constructed a dual tunable system for balancing the expression of CYP5150L8 and a Ganoderma P450 reductase iGLCPR, and performed a comprehensive optimization of CYP5150L8 expression, iGLCPR expression, and glycerol usage. Then, we investigated the fermentation behavior of the best strain in optimized condition in flask and achieved 154.45 mg/L GA-HLDOA production, which was 10.7-fold higher compared with previous report. This study may facilitate the wide-spread application of GA-HLDOA and the discovery of unknown cytochrome P450s in downstream GAs biosynthesis.     

Fungal degradation of lantadene A, the pentacyclic triterpenoid hepatotoxin of lantana plant

Seven white-rot fungi, known to degrade complex biomolecules and xenobiotics were used for investigation on biodegradation of lantadene A (LA), the bioactive pentacyclic triterpenoid of Lantana camara. Pleurotus sajor-caju and Merulius tremellosus PRL-2845 did not degrade LA. Trametes versicolor MTCC-138, Heterobasidion annosum MTCC-146, Phellinus pini RAB-83-19, Pleurotus ostreatus MTCC-142 and Phlebia radiata 2 utilized LA to the extent of 11–15.7%. This is the first report on the fungal degradation of a pentacyclic triterpenoid.     

灵芝酸A对前列腺癌LNCaP细胞的生长抑制作用及机制

本研究利用alamarBlue~?测定细胞活力法、流式细胞术(annxin V-FITC)/PI双染色测定细胞凋亡法和高效液相色谱(high performance liquid chromatography,HPLC)测定5α-还原酶活性法评价灵芝酸A对前列腺癌的体外抗肿瘤活性,并进一步利用流式细胞术2’,7’-二氯荧光素二乙酸酯(2’,7’-dichlorofluorescindiacetate,H2DCFDA)染色测定ROS释放量法、实时荧光定量PCR(quantitative real-time PCR,qPCR)和蛋白质免疫印迹(western blot,WB)检测雄激素受体(androgen receptor,AR)基因和凋亡相关基因表达的方法,探讨其作用机理。研究结果表明灵芝酸A可通过抑制5α-还原酶活性,抑制睾酮诱导的前列腺癌LNCaP细胞增殖,诱发细胞早期凋亡来发挥抗肿瘤活性;进一步研究表明,灵芝酸A降低前列腺癌细胞中AR的表达,并通过引发细胞线粒体功能障碍释放过量活性氧(reactive oxygen species,ROS)诱发细胞凋亡,而qPCR和WB的数据进一步表明细胞线粒体功能障碍与抑癌基因caspase-3、bad和aifm1的高表达密切相关。     

Schizonellin A and B,new glycolipids from Schizonella melanogramma

Two new glycolipids, schizonellin A and B, were isolated from cultures of the smut fungus Schizonella melanogramma. Their structures were elucidated by chemical degradation and spectroscopic methods. The schizonellins are active against Gram-positive bacteria, as well as against some Gram-negative bacteria and some fungi. DNA-, RNA- and protein-syntheses in Ehrlich carcinoma ascitic cells are inhibited simultaneously after addition of the schizonellins. Strong haemolytic action on bovine erythrocytes was observed.     

Aristoloside,an aristolochic acid derivative from stems of Aristolochia manshuriensis

Aristoloside, a new companion aristolochic acid derivative isolated from stems of Aristolochia manshuriensis has been shown to be 6-O-β-d-glucopyranoside of aristolochic acid-D on chemical and physicochemical evidence. Three known acids, aristolochic acids I, IV (both as their corresponding methyl esters), and -D have also been characterized from stems of the plant.