Insights into Triterpene Acids in Fermented Mycelia of Edible Fungus Poria cocos by a Comparative Study.

As an edible sclerotia-forming fungus, Poria cocos is widely used as a food supplement and as a tonic in China. High-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (HPLC-QTOF-MS/MS) was applied to identify triterpene acids in fermented mycelia of P. cocos, as well as the epidermis and inner part of natural sclerotia. A total of 19 triterpene acids were identified in fermented mycelia, whereas 31 were identified in the epidermis and 24 in the inner part. Nine triterpene acids were quantitatively determined, and the concentrations of two valuable triterpenes, dehydropachymic acid and pachymic acid, reached 1.07 mg/g and 0.61 mg/g in the fermented mycelia part, respectively, and were both significantly higher than the concentration in the two natural parts. The fermented mycelia could be a good choice for producing some target triterpene compounds and functional foods through fermentation thanks to the high concentration of some triterpene acids.

Supplementation with compound polysaccharides contributes to the development and metabolic activity of young rat intestinal microbiota.

Dietary intervention during early life has a significant impact on colonization of the gut microbiota. In addition, some polysaccharides have the potential to selectively stimulate the growth and metabolic activity of intestinal bacteria associated with health and well-being. However, less is known about the effect of polysaccharides on the development of gut microbiota in younger individuals. This study was conducted to investigate the health effects of supplementation with dietary compound polysaccharides (Lycium barbarum polysaccharides (LBP), Poria cocos polysaccharides (PCPs) and Lentinan, 1 : 1 : 1) on the intestinal microecosystem and metabolism of young rats. Male 21-day-old Sprague-Dawley rats received daily intragastric administration of either compound polysaccharides (three dosages, 6 g kg-1, 12 g kg-1 or 24 g kg-1) or saline for 28 consecutive days. 1H-NMR spectroscopy integrated with multi-variate pattern recognition analysis was applied to reveal the metabolism of the host and microflora, while 16S rRNA gene sequencing was used to monitor the dynamic changes in the gut microbiota. The relative concentrations of 35 urinary metabolites and 24 faecal metabolites were significantly changed compared with the control group. 16S rRNA analysis showed that the relative abundances of 4 bacterial genera (Bifidobacterium, Lactobacillus, Allobaculum and Oligella) significantly increased, whereas the relative abundance of 1 bacterial genus (Enterococcus) significantly declined in the compound polysaccharide-treated groups compared with the control group. Meanwhile, dietary compound polysaccharide treatment promoted the functional maturation of the gut bacterial community, characterised by increased basic metabolism (amino acid metabolism and energy metabolism), short chain fatty acid (SCFA)-related metabolism and nucleotide metabolism. These findings suggest that compound polysaccharides may help to promote the colonisation and functional maturation of infant intestinal microbiota and maintain the health of the intestinal microecosystem.

Targeted Screening Approach to Systematically Identify the Absorbed Effect Substances of Poria cocos in Vivo Using Ultrahigh Performance Liquid Chromatography Tandem Mass Spectrometry.

Poria cocos are extensively used as nutritious food, dietary supplements, and oriental medicine in Asia. However, the effect substances are still not very clear. In this study, a targeted screening approach was developed to systematically identify absorbed constituents of Poria cocos in vivo using ultrahigh performance liquid chromatography tandem mass spectrometry combined with UNIFI software. First, incubation reactions in vitro with rat intestinal microflora and rat liver microsomes were conducted to sum up metabolic rules of main constituents. Second, the absorbed constituents in vivo were picked out and identified based on the results of metabolic study in vitro. Finally, the absorbed active constituents in the treatment of Alzheimer's disease were screened by targeted network pharmacology analysis. A total of 62 absorbed prototypes and 59 metabolites were identified and characterized in dosed plasma. Thirty potential active constituents were screened, and 86 drug-targets shared by absorbed constituents and Alzheimer's disease were discovered by targeted network pharmacology analysis. In general, this proposed targeted strategy comprehensively provides new insight for active ingredients of Poria cocos.

Pachymic acid protects H9c2 cardiomyocytes from lipopolysaccharide-induced inflammation and apoptosis by inhibiting the extracellular signal-regulated kinase 1/2 and p38 pathways.

Pachymic acid (PA), a lanostane-type triterpenoid and the major component of Poria cocos alcoholic extracts, has various pharmacological effects, including anti-inflammatory, anti-oxidative and anti-apoptotic. However, few studies have investigated the effects of PA on lipopolysaccharide (LPS)-induced H9c2 cell apoptosis and inflammation, or identified the possible mechanisms underlying these effects. In the present study, H9c2 cardiomyocytes were stimulated by LPS and treated with or without PA. The increased mRNA expression levels of tumor necrosis factor-α, interleukin (IL)-1 and IL-6 induced by LPS were attenuated following treatment with PA. PA also attenuated LPS-induced apoptosis, as determined by a terminal deoxynucleotidyl transferase dUTP nick end labeling assay, and regulated the LPS-induced protein expression levels of caspase 3, 8 and 9. Furthermore, the phosphorylations of extracellular-regulated kinase (Erk)1/2 and p38 in the LPS-treated H9c2 cells were inhibited by PA. These results suggested that treatment with PA prevented the LPS-induced inflammatory and apoptotic response in cardiomyocytes, which may be mediated by inhibition of the Erk1/2 and p38 pathways.

[Effects of Oxygen Carriers on Submerged Fermentation of Poria cocos].

OBJECTIVE: To explore the regulation effects of oxygen carriers on Poria cocos submerged fermentation system which usually can be seriously inhibited by dissolved oxygen limitation. METHODS: One-factor-at-a-time design was employed to determine the oxygen carrier addition strategy through analyzing the effects of different oxygen carries, concentration and adding time of oxygen carrier on Poria cocos submerged fermentation. Then the oxygen carrier addition strategy was established and the metabolic processes of Poria cocos submerged fermentation were investigated comprehensively. RESULTS: The optimal oxygen carrier addition strategy was adding 1% (V/V) Tween-80 at 48 h after inoculation. Under this optimized condition, dry cell weight of Poria cocos reached 13.43 g/L in a 10 L bioreactor, while yields of exopolysaccharides and pachymic acid were 8.58 g/L and 989.52 µg/L, respectively, which exhibited obvious promoting effects compared with no addition oxygen carrier fermentation process. CONCLUSION: Tween-80 can remarkably increase the levels of cell growth, exopolysaccharides biosynthesis and pachymic acid in Poria cocos submerged fermentation system, which may provide new reference for further exploring dissolved oxygen limitation in high density fermentation of medical fungi efficiently.

Cloning and characterization of squalene synthase gene from Poria cocos and its up-regulation by methyl jasmonate.

Squalene synthase (SQS) catalyzes the condensation of two molecules of farnesyl diphosphate to give presqualene diphosphate and the subsequent rearrangement to form squalene. The gene encoding squalene synthase was cloned from Poria cocos by degenerate PCR and inverse PCR. The open reading frame of the gene is 1,497 bp, which encodes 499 amino acid residues. A phylogenetic analysis revealed that P. cocos SQS belonged to the fungus group, and was more closely related to the SQS of Ganoderma lucidum than other fungi. The treatment of P. cocos with methyl jasmonate (MeJA) significantly enhanced the transcriptional level of P. cocos sqs gene and the content of squalene in P. cocos. The transcriptional level of sqs gene was approximately fourfold higher than the control sample and the squalene content reached 128.62 µg/g, when the concentration of MeJA was 300 µM after 72 h induction.

Removal of copper, chromium and arsenic from preservative-treated wood by chemical extraction-fungal bioleaching.

Large volumes of preservative-treated wood containing toxic Cr, Cu and As salts are decommissioned worldwide. This study investigated the effectiveness of solid-state fermentation with copper-tolerant brown-rot fungi for the remediation of wood treated with chromated copper arsenate (CCA) and acid copper chromate (ACC) formulations. Treatment of CCA- and ACC-wood with the most effective strain, Antrodia vaillantii FRLP-14G, attained extensive leaching of As and/or Cr, but Cu elimination was poor (<18%). Additional research showed that a variety of organic acids, including citrate, are effective Cu extractants. Based on these findings, a process combining chemical extraction and subsequent fungal treatment was developed that proved highly effective in removing inorganic pollutants from CCA-wood. Extraction of CCA-wood with citric acid (30 mM, pH 3.10) followed by a 28-day solid-state fermentation period removed 87% Cu, 80% Cr, and 100% As. These results indicate the potential of the two-stage process for the remediation of preservative-treated wood.