Search

To systemically understand the biosynthetic pathways of bioactive substances, including triterpenoids and polysaccharides, in Ganoderma lucidum, the correlation between substrate degradation and carbohydrate and triterpenoid metabolism during growth was analyzed by combining changes in metabolite content and changes in related enzyme expression in G. lucidum over 5 growth phases. Changes in low-polarity triterpenoid content were correlated with changes in glucose and mannitol contents in fruiting bodies. Additionally, changes in medium-polarity triterpenoid content were correlated with changes in the lignocellulose content of the substrate and with the glucose, trehalose, and mannitol contents of fruiting bodies. Weighted gene coexpression network analysis (WGCNA) indicated that changes in trehalose and polyol contents were related to carbohydrate catabolism and polysaccharide synthesis. Changes in triterpenoid content were related to expression of the carbohydrate catabolic enzymes laccase, cellulase, hemicellulase, and polysaccharide synthase and to the expression of several cytochrome P450 monooxygenases (CYPs). It was concluded that the products of cellulose and hemicellulose degradation participate in polyol, trehalose, and polysaccharide synthesis during initial fruiting body formation. These carbohydrates accumulate in the early phase of fruiting body formation and are utilized when the fruiting bodies mature and a large number of spores are ejected. An increase in carbohydrate metabolism provides additional precursors for the synthesis of triterpenoids. IMPORTANCE Most studies of G. lucidum have focused on its medicinal function and on the mechanism of its activity, whereas the physiological metabolism and synthesis of bioactive substances during the growth of this species have been less studied. Therefore, theoretical guidance for cultivation methods to increase the production of bioactive compounds remains lacking. This study integrated changes in the lignocellulose, carbohydrate, and triterpenoid contents of G. lucidum with enzyme expression from transcriptomics data using WGCNA. The findings helped us better understand the connections between substrate utilization and the synthesis of polysaccharides and triterpenoids during the cultivation cycle of G. lucidum . The results of WGCNA suggest that the synthesis of triterpenoids can be enhanced not only through regulating the expression of enzymes in the triterpenoid pathway, but also through regulating carbohydrate metabolism and substrate degradation. This study provides a potential approach and identifies enzymes that can be targeted to regulate lignocellulose degradation and accelerate the accumulation of bioactive substances by regulating substrate degradation in G. lucidum .

Two polysaccharide fractions (GLSB50 and GLSB70) with total sugar content of 82.07 wt% and 53.79 wt%, respectively, were obtained from the water extracts of unbroken Ganoderma lucidum spores by sequential ethanol precipitation treatment. Compared with GLSB70, GLSB50 exhibited better activity on stimulation of humoral immune responses in immunosuppressed mice. A novel β-D-glucan (GLSB50A-III-1) with weight average molecular weight (M w ) of 1.93 × 10 5  g/mol was purified from GLSB50 through chromatography separation. The exponent α value of Mark-Houwink-Sakurada equation was calculated to be 0.13, indicating that GLSB50A-III-1 presented globular spheres conformation in aqueous solution. Structural analysis showed that GLSB50A-III-1 mainly consisted of (1 → 3), (1 → 4), (1 → 6)-linked β-d-glucose residues in the backbone, with two single β-D-Glcp attached at O-6 of β-(1 → 3) and β-(1 → 4)-linked residues separately as side chains. The repeat unit of GLSB50A-III-1 was deduced as follows., (Copyright © 2021. Published by Elsevier B.V.)

Ganoderma lucidum mycelia are rich in active substances such as triterpenoids and sterols. However, reports on the development of effective submerged fermentation processes are lacking and the resulting total triterpene and sterol yield is still quite low. In this study, a new G. lucidum strain G0017 mycelium isolated by screening was studied in a 3-L fermenter to investigate the effect of aeration rate in liquid submerged fermentation production of triterpenoids and sterols. By fitting the specific mycelial growth rate and the specific production rate of the triterpenoid and sterol model, an effective multistage aeration rate control process for triterpenoid and sterol fermentation production was developed. This process was validated and proven in 3-L and 50-L fermenters. The resulting yields of triterpenoids and sterols were 3.34 and 3.46 g/L, respectively, which were 69.54% and 75.63% higher than the fixed aeration rate of 1.50 volume of air per volume of liquid per minute. This optimized fermentation production process conceivably could be applied to larger-scale industrial production and perhaps also to improve liquid submerged fermentation processes with relevant edible and medicinal mushrooms.

In the present study, regioselective sulfation of β-glucan (GLP) from Ganoderma lucidum were firstly established by using 4,4'-dimethoxytrityl chloride and hexamethyldisilazane as protecting precursor. 2,4,6-O-sulfated, 6-O-sulfated and 2,4-O-sulfated GLP derivatives were prepared and the molecular weights (M w ) of derivatives were determined to range from 0.94 × 10 4 to 6.27 × 10 4  g/mol, while the degrees of sulfation (DS) were calculated to vary from 0.83 to 1.74. The regioselective sulfation of GLP was confirmed by FT-IR, 13 C NMR spectroscopy and methylation analysis. Results indicated that the sulfated substitution sites were predominantly at C-6 in 6-O-sulfated GLP (S 6-O GLP) and C-4 in 2,4-O-sulfated GLP (S 2,4-O GLP), respectively. Clotting assays (APTT, PT and TT) in vitro showed that sulfate groups were essential for anticoagulant activity and S 6-O GLP exhibited much higher than others. Meanwhile, sulfated GLP with higher DS and M w showed stronger anticoagulant activity in the case of the same condition., Competing Interests: Declaration of competing interest The authors declare no competing financial interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The mushroom today known as Ganoderma lingzhi has been used for centuries in the countries of Eastern Asia as a very important medicinal mushroom. It prefers growing on rotten wood of broadleaf trees and is mainly distributed in the tropics and subtropics. Its relative G. lucidum occurs naturally almost all the Earth, and it colonizes mostly oak and beech trees in Central Europe. G. lingzhi and G. lucidum are similar species. To obtain the qualitative parameters of G. lingzhi and G. lucidum, several strains (five G. lingzhi strains and five G. lucidum ones) were chosen and cultivated in both Slovakia and China, using wood chip (beech and oak) substrate and liquid fermentation method, respectively. It was found that there were more low-polarity triterpenes in G. lucidum, while G. lingzhi contained more high-polarity triterpenes. Beech substrate was more suitable for the accumulation of triterpenes in solid cultivation for both strains of G. lucidum and G. lingzhi. Strain C4 of G. lingzhi and strain K2 of G. lucidum contained higher triterpenes in either mycelium or fruiting bodies. Data in this study can help to identify these two species and bring a great benefit to the production of bioactive compounds of G. lucidum from Slovakia.

Sulfated polysaccharides exhibit various biological properties, including anti-coagulant, anti-oxidant, anti-viral, anti-cancer, anti-inflammatory and immune regulatory activities. In the present study, the anti-inflammatory properties of GLPss58, a sulfated polysaccharide from Ganoderma lucidum formed by chemical sulfation, were investigated. We found that GLPss58 inhibited L-selectin/sTyr-sLeX binding significantly, blocked the binding of anti-l-selectin antibodies to L-selectin on the surface of human peripheral blood lymphocytes, and inhibited the secondary lymphoid tissue chemokine-induced chemotactic invasion of HPBLs. In vivo studies in mice showed that lymphocyte homing from peripheral blood to spleen and lymph nodes was significantly inhibited by GLPss58. Furthermore, GLPss58 also inhibited the activation of complement systems and blocked the binding of TNF-α and IFN-γ to their antibodies. These results indicate that GLPss58 is able to inhibit not only the L-selectin-mediated inflammation, but also the complement system- and cytokines mediated-inflammation. Our results suggest that GLPss58 is a favorable potential anti-inflammatory agent., (Copyright © 2017 Elsevier B.V. All rights reserved.)

We analyzed the changes in triterpenes and soluble polysaccharides in Ganoderma lucidum strain G0119 during 4 growth phases in 3 regions of the fruiting bodies using reversed-phase high-performance liquid chromatography, and we also analyzed the soluble polysaccharides using high-performance size-exclusion chroma-tography-multiple-angle laser-light scattering refractive index analysis. The strong polar triterpenes decreased while weak polar triterpenes increased during the growth cycle of G. lucidum. The highest contents of ganoderic acid B, ganoderic acid A, and ganoderenic acid B were detected in the stipe during phase II, and ganoderic acid S, ganoderic acid T, and ganoderiol B peaked in the base during phase IV. The total content of soluble polysaccharides in samples decreased after the primordium developed into a fruiting body. Two high-molecular-weight fractions were detected in the soluble polysaccharide samples: α-l,4-glucan and β-l,3-glucan, respectively. They were primarily distributed in the pileus during phase II, and both decreased after this phase. These results led us to select a more suitable growth phase and region for harvesting to obtain extracts with higher contents of triterpenes and soluble polysaccharides.

A water soluble β-glucan (GLSWA-I) with a weight average molecular weight ∼1.57×10 5 g/mol, isolated from the aqueous extract of the broken cellular wall Ganoderma lucidum spores, was purified by anion-exchange and gel-permeation chromatography. The immunological activity test in vivo showed that GLSWA-I could significantly promote dinitrochlorobenzene (DNCB) induced delayed-type ear swelling in mice. Based on the monosaccharides composition analysis, methylation analysis, IR, 1D and 2D NMR spectroscopy, the repeating unit of polysaccharide GLSWA-I was elucidated as follows., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The immunomodulatory effect of GLIS (Lingzhi or Reishi medicinal mushroom Ganoderma lucidum immunomodulating substance) on macrophages has been investigated as part of ongoing research into the anticancer properties of this mushroom. Proliferation of bone marrow macrophages (BMMs) was enhanced by GLIS in a dose-dependent manner. Microscopic examination revealed that numerous GLIS-treated BMMs were enlarged and formed pseudopodia. Exposure of BMMs to GLIS resulted in significant increases in NO production, induction of cellular respiratory burst activity, and increased levels of IL-1β, IL-6, IL-12p35, IL-12p40, IL-18, and TNF-α gene expression and levels of TNF-α, IL-1β, and IL-12 secretion. Our data indicate that GLIS activates the immune system by modulating cytokine production.

A water-soluble glycopeptide (GLPCW-II) was isolated from the fruiting bodies of Ganoderma lucidum by DEAE-Sepharose Fast-Flow and Sephacryl S-300 High Resolution Chromatography. The glycopeptide had a molecular weight of 1.2x10(4)Da (determined by HPLC), and consisted of approximately 90% carbohydrate and approximately 8% protein as determined using the phenol-sulfuric acid method and the BCA protein assay reagent kit, respectively. The polysaccharide moiety was composed mainly of D-Glc, L-Fuc, and D-Gal in the ratio of 1.00:1.09:4.09. To facilitate structure-activity studies, the structure of the GLPCW-II polysaccharide moiety was elucidated using 1H and 13C NMR spectroscopy including COSY, TOCSY, HMBC, HSQC, and ROESY, combined with GC-MS of methylated derivatives, and shown to consist of repeating units with the following structure: [Formula: see text].

The immunomodulatory effect of Ganoderma lucidum immunomodulating substance (GLIS) on macrophages has been investigated as part of on-going research into the anti-cancer properties of Ganoderma lucidum. Proliferation of bone marrow macrophages (BMMs) was enhanced by GLIS in a dose-dependent manner. Microscopic examination revealed that numerous GLIS-treated RAW264.7 macrophages were enlarged and formed pseudopodia. Exposure of RAW264.7 macrophages to GLIS resulted in significant increases in NO production, induction of cellular respiratory burst activity, and increased levels of IL-1beta, IL-12p35 and IL-12p40 gene expression. Our data indicate that GLIS activates the immune system by modulating cytokine production.