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Khz (Fusion of Ganoderma lucidum and Polyporus umbellatus Mycelia) Induces Apoptosis by Increasing Intracellular Calcium Levels and Activating JNK and NADPH Oxidase-Dependent Generation of Reactive Oxygen Species  [PDF]
Tae Hwan Kim, Ju sung Kim, Zoo haye Kim, Ren Bin Huang, Ren Sheng Wang
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0046208
Abstract: Khz is a compound derived from the fusion of Ganoderma lucidum and Polyporus umbellatus mycelia that inhibits the growth of cancer cells. The results of the present study show that Khz induced apoptosis preferentially in transformed cells and had only minimal effects on non-transformed cells. Furthermore, Khz induced apoptosis by increasing the intracellular Ca2+ concentration ([Ca2+]i) and activating JNK to generate reactive oxygen species (ROS) via NADPH oxidase and the mitochondria. Khz-induced apoptosis was caspase-dependent and occurred via a mitochondrial pathway. ROS generation by NADPH oxidase was critical for Khz-induced apoptosis, and although mitochondrial ROS production was also required, it appeared to occur secondary to ROS generation by NADPH oxidase. Activation of NADPH oxidase was demonstrated by the translocation of regulatory subunits p47phox and p67phox to the cell membrane and was necessary for ROS generation by Khz. Khz triggered a rapid and sustained increase in [Ca2+]i, which activated JNK. JNK plays a key role in the activation of NADPH oxidase because inhibition of its expression or activity abrogated membrane translocation of the p47phox and p67phox subunits and ROS generation. In summary, these data indicate that Khz preferentially induces apoptosis in cancer cells, and the signaling mechanisms involve an increase in [Ca2+]i, JNK activation, and ROS generation via NADPH oxidase and mitochondria.
Effect of the Mycelial Culture of Ganoderma lucidum on Human Pathogenic Bacteria
Lauretta Nwanneka Ofodile,A.O. Ogbe,Olufunke Oladipupo
International Journal of Biology , 2011, DOI: 10.5539/ijb.v3n2p111
Abstract: The mycelial culture of Ganoderma lucidum found in Nigeria was tested on some human pathogenic bacteria using dual and Weller techniques. The mycelial culture was active against all the test organisms: Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae and Pseudomonas aeruginosa. The bacterium, Escherichia coli showed the highest susceptibility to the mushroom mycelia at 10.6 mm on the first day of incubation which increased to 15.2 mm on the fourth day using the dual method while Pseudomonas aeruginosa showed the least susceptibility of 5.7 mm on the fourth day. In the Weller method the growth of the test organisms were completely antagonized by the mycelia of Ganoderma lucidum as total inhibition was recorded from all the plates. These could be an indication that Ganoderma lucidum can be used in the treatment of diseases caused by these organisms as well as preventive measure against them.
Study of different growth parameters in ganoderma lucidum  [cached]
Z. Nasreen,T. Kausar,M. Nadeem,R. Bajwa
Micología Aplicada Internacional , 2005,
Abstract: Ganoderma lucidum was grown in several culture media, using four different concentrations of inoculum. The mycelia grew well in the potato dextrose medium at pH 5 and 25 C, followed by malt extract, Kirk medium + molasses, and Kirk medium + glucose. The yield of mycelial biomass (dry weight basis) in potato dextrose medium after 15 days was about 1.59 g/100 ml; in malt extract, 0.91 g/100 ml; in Kirk molasses, 0.68 g/100 ml; and in Kirk glucose was 0.38 g/100 ml. Mycelial growth of G. lucidum was significantly higher in case of 5 to 15 ml inoculum concentration.
Complete Mitochondrial Genome of the Medicinal Mushroom Ganoderma lucidum  [PDF]
Jianqin Li, Jianhui Zhang, Haimei Chen, Xiangdong Chen, Jin Lan, Chang Liu
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0072038
Abstract: Ganoderma lucidum is one of the well-known medicinal basidiomycetes worldwide. The mitochondrion, referred to as the second genome, is an organelle found in most eukaryotic cells and participates in critical cellular functions. Elucidating the structure and function of this genome is important to understand completely the genetic contents of G. lucidum. In this study, we assembled the mitochondrial genome of G. lucidum and analyzed the differential expressions of its encoded genes across three developmental stages. The mitochondrial genome is a typical circular DNA molecule of 60,630 bp with a GC content of 26.67%. Genome annotation identified genes that encode 15 conserved proteins, 27 tRNAs, small and large rRNAs, four homing endonucleases, and two hypothetical proteins. Except for genes encoding trnW and two hypothetical proteins, all genes were located on the positive strand. For the repeat structure analysis, eight forward, two inverted, and three tandem repeats were detected. A pair of fragments with a total length around 5.5 kb was found in both the nuclear and mitochondrial genomes, which suggests the possible transfer of DNA sequences between two genomes. RNA-Seq data for samples derived from three stages, namely, mycelia, primordia, and fruiting bodies, were mapped to the mitochondrial genome and qualified. The protein-coding genes were expressed higher in mycelia or primordial stages compared with those in the fruiting bodies. The rRNA abundances were significantly higher in all three stages. Two regions were transcribed but did not contain any identified protein or tRNA genes. Furthermore, three RNA-editing sites were detected. Genome synteny analysis showed that significant genome rearrangements occurred in the mitochondrial genomes. This study provides valuable information on the gene contents of the mitochondrial genome and their differential expressions at various developmental stages of G. lucidum. The results contribute to the understanding of the functions and evolution of fungal mitochondrial DNA.

ZHANG Jin-Song JIA Wei XING Zhen-Tao TANG Qing-Jiu LIU Yan-Fang,YANG Yan ZHOU Chang-Yan LIU Fang,

菌物学报 , 2004,
Abstract: 从不同品种的灵芝中筛选出了多糖含量最高的菌株GL2为材料,利用柱层析技术从子实体和菌丝体提取物中分离得到多个组分。实验发现子实体组分P3,P31及P32对人白血病细胞株K562的生长有明显地抑制作用,这三个组分中只有P32对另一白血病细胞株HL-60有抑制作用。免疫活性测试的结果显示子实体各组分在刺激小鼠脾淋巴细胞,T和B细胞的增殖,提高人外周血中NK细胞杀伤活性方面比菌丝体的作用强;进一步的实验发现子实体与菌丝体相应组分在刺激人外周血中的T和B淋巴细胞增殖方面的活性差异不大。子实体和菌丝体提取物各组份均可剂量依赖型的促进PBMC分泌TNF-α。菌丝体提取物各组分对TNF-释放量的影响在低浓度时与子实体各组份相当,在高浓度时要明显好于赤芝子实体提取物各组分。
Purification of an Intracellular Fibrinolytic Protease from Ganoderma Lucidum Vk12 and its Susceptibility to Different Enzyme Inhibitors
S Kumaran, P Palani, R Nishanthi, S Srimathi, V Kaviyarasan
Tropical Journal of Pharmaceutical Research , 2011,
Abstract: Purpose: To study the effect of different inhibitors on the fibrinolytic activity of the enzyme produced by Ganoderma lucidum. Method: The intracellular fibrinolytic protease produced by Ganoderma lucidum VK12 was isolated from the mycelia grown in MCDBF broth and was purified to homogeneity using ammonium sulfate fractionation, ion exchange chromatography and sephadex G-150 column chromatography techniques. The purity of the enzyme was verified on SDS-PAGE after silver nitrate staining. The inhibitory effect of different metal ions and commercial protease inhibitors on enzyme activity was studied. The inhibitortreated enzyme was assayed with its substrate and the residual activity of the enzyme recorded. Result: The fibrinolytic enzyme isolated from Ganoderma lucidum was purified to near homogeneity and it appeared as a single protein band on SDS-PAGE. Metal ions such as Ca2+ and Mg2+ inhibited the activity of the enzyme while Zn2+ ions enhanced the activity. Complete inactivation occurred when the enzyme was incubated with protease inhibitors such as EDTA, 1, 10-phenanthroline, phosphoamidon while the enzyme was insensitive to protease inhibitors such as leupeptin, PMSF, TPCK and APMSF. Conclusion: Copper sulfate completely inhibited the enzyme activity. The sensitivity of this enzyme to EDTA suggests that it might be a metalloprotease.

YANG Xiao-Tong ZHOU Yue-Qin LI Xu-Quan FENG Hui-Qin MI Ke YANG Qing-Yao,

菌物学报 , 2005,
Abstract: 以人急性早幼粒白血病细胞HL-60细胞株作为模型,用MTT法测定生长抑制率,流式细胞Annexin Ⅴ实验和DNA含量法研究抑瘤机制,比较了8个灵芝Ganoderma lucidum菌株发酵菌丝体乙醇提取物的体外抗肿瘤活性和抑瘤机制。筛选结果得到了能产生高抑瘤活性乙醇提取物的灵芝菌株L5。其对HL-60细胞的抑制率为91.4±0.9%(72小时,125μg/mL); 作用48小时后,13.3%的细胞发生早期凋亡,G0/G1期细胞比例比对照组增加15.9%,而S期细胞比例则下降了8.4%,G2/M期细胞数减少7.6%。本研究证明了灵芝菌丝体乙醇提取物能有效抑制HL-60肿瘤细胞的体外生长,抑制率与菌株相关,其抑瘤机制与细胞G0/G1期阻滞和诱导凋亡有关。
Evaluation of Antibacterial activity of Polysaccharide Extract of Ganoderma lucidum  [cached]
Priya Batra and Robinka Khajuria
Current Trends in Biotechnology and Chemical Research , 2012,
Abstract: The use of natural products and herbal medicines is becoming popular due to the recognition of value of traditional medical system and the identification of indigenous medicinal plants. Furthermore, there is a need of constant search for new and effective drugs due the emergence of resistant strains against currently used drugs. Ganoderma Species are one of the most widely researched medicinal mushrooms because of their potent bioactive properties. Therefore this study aims to evaluate the antibacterial potential of Ganoderma lucidum. Hot water extract of G. lucidum was used to obtain the polysaccharide fraction by ethanol precipitation. The antibacterial activity was analysed by disc diffusion method and spectrophotometeric analysis. The largest zone of inhibition (19mm) was observed for Staphylococcus sp. followed by Pseudomonas aeruginosa, Salmonella typhi, Klebsiella sp. And Escherichia coli. A decrease of about 70% was observed in the growth rate of Staphylococcus sp. when incubated in the presence of G. lucidum aqueous extract, followed by E. coli (40%), Klebsiella sp. (26%), P. aeruginosa (21%) and S. typhi (12%).
Deep Insight into the Ganoderma lucidum by Comprehensive Analysis of Its Transcriptome  [PDF]
Guo-Jun Yu, Man Wang, Jie Huang, Ya-Lin Yin, Yi-Jie Chen, Shuai Jiang, Yan-Xia Jin, Xian-Qing Lan, Barry Hon Cheung Wong, Yi Liang, Hui Sun
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0044031
Abstract: Background Ganoderma lucidum is a basidiomycete white rot fungus and is of medicinal importance in China, Japan and other countries in the Asiatic region. To date, much research has been performed in identifying the medicinal ingredients in Ganoderma lucidum. Despite its important therapeutic effects in disease, little is known about Ganoderma lucidum at the genomic level. In order to gain a molecular understanding of this fungus, we utilized Illumina high-throughput technology to sequence and analyze the transcriptome of Ganoderma lucidum. Methodology/Principal Findings We obtained 6,439,690 and 6,416,670 high-quality reads from the mycelium and fruiting body of Ganoderma lucidum, and these were assembled to form 18,892 and 27,408 unigenes, respectively. A similarity search was performed against the NCBI non-redundant nucleotide database and a customized database composed of five fungal genomes. 11,098 and 8, 775 unigenes were matched to the NCBI non-redundant nucleotide database and our customized database, respectively. All unigenes were subjected to annotation by Gene Ontology, Eukaryotic Orthologous Group terms and Kyoto Encyclopedia of Genes and Genomes. Differentially expressed genes from the Ganoderma lucidum mycelium and fruiting body stage were analyzed, resulting in the identification of 13 unigenes which are involved in the terpenoid backbone biosynthesis pathway. Quantitative real-time PCR was used to confirm the expression levels of these unigenes. Ganoderma lucidum was also studied for wood degrading activity and a total of 22 putative FOLymes (fungal oxidative lignin enzymes) and 120 CAZymes (carbohydrate-active enzymes) were predicted from our Ganoderma lucidum transcriptome. Conclusions Our study provides comprehensive gene expression information on Ganoderma lucidum at the transcriptional level, which will form the foundation for functional genomics studies in this fungus. The use of Illumina sequencing technology has made de novo transcriptome assembly and gene expression analysis possible in species that lack full genome information.
Use of RAPD molecular markers on differentiation of brazilian and chinese Ganoderma lucidum strains
Rolim, Leonardo do Nascimento;Cavalcante, Maria Auxiliadora de Queiroz;Urben, Arailde Fontes;Buso, Glaucia Salles Cortopassi;
Brazilian Archives of Biology and Technology , 2011, DOI: 10.1590/S1516-89132011000200008
Abstract: the aim of this work was to analyze the brazilian and chinese strains of ganoderma lucidum with molecular rapd markers. a similarity matrix was elaborated and the rapd profiles of g. lucidum strains were also compared to two other ganoderma spp: g. applanatum and g. lipsiense in order to produce genetic similarity among the species. based on the primers used, it was possible to determine that the brazilian strains and chinese strain cc-22 are alike. the method and the primers selection showed to be appropriate for the genetic identification of g. lucidum strains, enabling them to be improved and used in research, as well as in the world market.
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