Ganoderma Tsugae Research Articles

Fresh-eating Lingzhi becomes possible: Comparative evaluation of nutritional and taste profile of Ganoderma tsugae at different fruiting body morphogenesis stages

Ganoderma tsugae is a significant mushroom with medicinal and dietary properties. However, it has not been used as dishes and fresh-eating food. In order to fully explore this rare mushroom resource, an in-depth investigation was conducted on the dynamic changes in various nutritional and taste indicators of G. tsugae for the first time. This study focused on an analysis of the nutritional components, mineral elements, non-volatile taste compounds, electronic tongue, and texture profile analysis of samples at six different morphogenesis stages (primordia stage, coloration stage, stipe differentiation stage, pileus differentiation stage, spore pre-ejection stage, late stage of spore ejection, respectively). The results indicated that the nutrient and mineral contents in primordia and coloration stage exhibited superior qualities compared to the other treatments. Partial least squares discriminant analysis (PLS-DA) and principal component analysis (PCA) revealed the differences and comprehensive evaluation in taste and nutritional value between different treatments. According to the analysis of the edibility and economic benefit of various samples, it has been determined that the optimal harvest stage for the fresh-eating G. tsugae is coloration stage. This finding serves as a fundamental basis for the development and utilization of fresh culinary lingzhi. It also provides essential data and a comprehensive reference for addressing the issue of raw materials in the production of Ganoderma products.

Antibacterial Lanostane Triterpenoids from the Fruiting Bodies of Ganoderma tsugae

Antibacterial Lanostane Triterpenoids from the Fruiting Bodies of Ganoderma tsugae

Antibacterial lanostane triterpenoids from Ganoderma tsugae

A phytochemical investigation on the 80% EtOH extract of the fruiting bodies of Ganoderma tsugae resulted into the isolation of two previously undescribed lanostane triterpenoids, 7,11-dioxo-3β-acetyloxy-26,27-dihydroxy-lanosta-8,24-diene (1) and 7,20-dioxo-3β-acetyloxy-11β,15α-dihydroxy-22,23,24,25,26,27-hexanorlanosta-8-ene (2), togeher with one known lanostane triterpenoid ganodermanontriol (3). Structural elucidation of all the compounds were performed by spectral methods such as 1D and 2D (1H- 1H COSY, HMQC, and HMBC) NMR spectroscopy. All the triterpenoids were in vitro evaluated for their antibacterial activities against six pathogenic microorganisms. Compound 3 exhibited some activities against three Gram positive bacteria with MIC values less than 30 μg/ml.

The Development of Novel Ganoderic-Acid-Encapsulated Nanodispersions Using the Combination of Ultrasonic Cavitation and Solvent Evaporation through Response Surface Optimization

Ganoderic Acid (GA), a major bioactive compound isolated from the East Asian medicinal mushroom Ganoderma tsugae, is traditionally believed to have significant medicinal properties. GA is poorly soluble in water, which poses several challenges in terms of its formulation. In this study, Ganoderma tsugae extracts obtained through ethanol extraction were encapsulated in nanodispersions via ultrasonic cavitation and solvent evaporation to increase their bioavailability. The preparation route was thoroughly analyzed using Response Surface Methodology (RSM) to determine the interactions between the variables. Based on the results, the Hydrophilic–Lipophilic Balance (HLB) and the evaporation temperature significantly influenced the resulting particle size. In the optimized nanodispersions, GA was incorporated into a hydrophobic core with a particle size no greater than 200 nm and a very narrow particle distribution (namely, a polydispersity index of 0.289). Due to the high negative zeta potential (−45.9 mV), a very slow particle growth rate of 0.239% over short-term storage (14 days) was achieved. In addition, the zeta average remained virtually unchanged for 14 days at room temperature in solutions at different pH levels. In summary, this paper provides the first-ever demonstration that ultrasound cavitation in conjunction with solvent evaporation can be used to generate GA nanodispersions.

Lanostane triterpenoids from the medicinal mushroom Ganoderma tsugae

Two undescribed lanostane triterpenoids, tsugaric acids G ( 1 ) and H ( 2 ), and a known compound, N -(3′ α ,4′ β -dihydroxy-2′ β -(hydroxymethyl)-1′ β -(cyclobutyl) palmitamide ( 3 ) were isolated from the fruiting bodies of Ganoderma tsugae and characterized on the basis of spectroscopic and spectrometric analysis, especially 2D NMR and HRMS. Compounds 1 and 2 were also characterized by an LC-ESI-MS analytical approach and were compared with the fragmentation patterns of reported compounds. Compound 3 significantly exhibited anti-HCV activity. • Two new lanostanoids ( 1 and 2 ) and one known palmitamide ( 3 ) were isolated from Ganoderma tsugae . • The new compounds were discovered by using an LC-ESI-MS analytical approach. • Compound 3 exhibited anti-HCV activity.

Hepatoprotective Activity of Ethanol Extract of Rice Solid-State Fermentation of Ganoderma tsugae against CCl4-Induced Acute Liver Injury in Mice.

Ganoderma tsugae is well known as a medicinal mushroom in China and many Asian countries, while its fermentation technique and corresponding pharmacological activity are rarely reported. In this study, a wild G. tsugae strain (G42) with high triterpenoid content was screened from nine strains by rice solid-state fermentation, and 53.86 mg/g triterpenoids could be produced under optimized conditions; that is, inoculation amount 20%, fermentation temperature 27 °C, and culture time 45 days. The hepatoprotective activity of G42 ethanol extract was evaluated by CCl4-induced liver injury in mice, in which changes in the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), oxidation-related factors, and inflammatory cytokines in serum or liver samples demonstrated the therapeutic effect. In addition, the ethanol extract of G42 reduced the incidence of necrosis and inflammatory infiltration, and decreased protein expression levels of phosphor-nuclear factor-κB (NF-κB), interleukin-Iβ (IL-1β), and nuclear factor erythroid-2-related factor 2 (NRF2). The chemical composition of the ethanol extract was analyzed by high-resolution mass spectrometry and molecular networking. Three main triterpenoids, namely platycodigenin, cucurbitacin IIb, and ganolecidic acid B were identified. This work provided an optimized fermentation method for G. tsugae, and demonstrated that its fermentation extract might be developed as a functional food with a hepatoprotective effect.

Ganoderma tsugae prevents cognitive impairment and attenuates oxidative damage in d-galactose-induced aging in the rat brain.

Lingzhi has long been regarded as having life-prolonging effects. Research in recent years has also reported that Lingzhi possesses anti-tumor, anti-inflammatory, immunomodulatory, hepatoprotective, and anti-lipogenic effects. The D-galactose (D-gal, 100 mg/kg/day)-induced aging Long-Evans rats were simultaneously orally administered a DMSO extract of Ganoderma tsugae (GTDE, 200 μg/kg/day) for 25 weeks to investigate the effects of GTDE on oxidative stress and memory deficits in the D-galactose-induced aging rats. We found that GTDE significantly improved the locomotion and spatial memory and learning in the aging rats. GTDE alleviated the aging-induced reduction of dendritic branching in neurons of the hippocampus and cerebral cortex. Immunoblotting revealed a significant increase in the protein expression levels of the superoxide dismutase-1 (SOD-1) and catalase, and the brain-derived neurotrophic factor (BDNF) in rats that received GTDE. D-gal-induced increase in the lipid peroxidation product 4-hydroxynonenal (4-HNE) was significantly attenuated after the administration of GTDE, and pyrin domain-containing 3 protein (NLRP3) revealed a significant decrease in NLRP3 expression after GTDE administration. Lastly, GTDE significantly reduced the advanced glycosylation end products (AGEs). In conclusion, GTDE increases antioxidant capacity and BDNF expression of the brain, protects the dendritic structure of neurons, and reduces aging-induced neuronal damage, thereby attenuating cognitive impairment caused by aging.

Kalappia celebica, an endemic wood from Sulawesi Island: Chemical composition and its resistance against white rot fungi Ganoderma tsugae and G. lucidium

Kalapi wood (Kalappia celebica kosterm) is endemic to Sulawesi Island and is a monotype species in the Fabaceae family with limited distribution. This study investigates the chemical composition and natural durability of Kalapi (K. celebica Kosterm) wood against white rot fungi Ganoderma tsugae and G. lucidium. This study was conducted based on the TAPPI standard for moisture analysis content, ASTM-D for extractive content, holocelluloce, and lignin, and method Cross and Bevan for cellulose. Durability observation was conducted in 12 weeks by assessing the percentage damage of wood samples caused by the fungi according to SNI standards 01-7207-2014. Results show that K. celebica has holocelullose, cellulose, and extractives at a high level, lignin was moderate, and had low hemicellulose. The results of testing the durability of wood shown that K. celebica has resistance to G. tsugae attack (class II) but is not resistant to G. lucidium (class IV), with an average weight loss of 3.14 % and 18.82 %, respectively.

Genome of Ganoderma Species Provides Insights Into the Evolution, Conifers Substrate Utilization, and Terpene Synthesis for Ganoderma tsugae.

Ganoderma tsugae is an endemic medicinal mushroom in Northeast China, providing important source of pharmaceutical product. Comparing with other Ganoderma species, wild G. tsugae can utilize coniferous wood. However, functional genes related to medicinal component synthesis and the genetic mechanism of conifer substrate utilization is still obscure. Here, we assembled a high-quality G. tsugae genome with 18 contigs and 98.5% BUSCO genes and performed the comparative genomics with other Ganoderma species. G. tsugae diverged from their common ancestor of G. lingzhi and G. sinense about 21 million years ago. Genes in G. tsugae-specific and G. tsugae-expanded gene families, such as salh, phea, cyp53a1, and cyp102a, and positively selected genes, such as glpk and amie, were functionally enriched in plant-pathogen interaction, benzoate degradation, and fanconi anemia pathway. Those functional genes might contribute to conifer substrate utilization of G. tsugae. Meanwhile, gene families in the terpene synthesis were identified and genome-wide SNP variants were detected in population. Finally, the study provided valuable genomic resources and offered useful hints for the functional gene mapping and investigation of key gene contributing to conifer cultivation substrate utilization and medicinal component biosynthesis.

Current Knowledge of Medicinal Mushrooms Related to Anti-Oxidant Properties

Background: A renewed focus on medicinal mushrooms has brought forth a sustainable health dimension. Conventional health strategies are insufficiently integrated with sustainable health promotion. The health-promoting outcome of mushrooms has fascinated many groups during the past few years because of various primary and secondary metabolites in different cellular components. They contain many bioactive metabolites, including proteins (cytokines, ergothioneine), fibers, moisture, carbohydrates (uronic acid), folate, thiamine, ascorbic acid, vitamin D, calcium, potassium, polysaccharides (G. lucidum polysaccharides, alpha and beta glucans, and lentinan) polyketides, polyphenols (Protocatechuic acid, inonoblins A–vanillic acid, phelligridins D, E, and G, hydroxybenzoic acid, gallic acid, tannic acid, hispidine, gentisic acid, and tocopherol), nucleotides (adenosine, cordycepin), lovastatin, steroids, alkaloids, and sesquiterpenes. Objective: This study was conducted to gather information on the current knowledge of medicinal mushrooms, with respect to their antioxidant properties. Conclusions: The results indicated that mushrooms are a promising source of natural antioxidants. Of all mushrooms, the Ganoderma tsugae Murill exhibited an excellent antioxidant potential of 93.7–100% at 20 mg/mL.

Degradation ability of wood decay fungi in teak (Tectona grandis L.) wood under storage condition

Aim: The present study was undertaken to provide basic knowledge about the teak woody biomass degradation ability of wood decay fungi. Methodology: Wood decay fungi was isolated from the decayed wood samples collected from teak wood logs. The weight loss of teak wood and moisture content were assessed under in-vitro condition by incubating the teak wood block with the isolated fungi. Further, the secretion of ligninolytic enzymes viz., laccase, lignin peroxidise (LiP) and manganese peroxidase (MnP) produced by wood decay fungi were assessed to understand their degrading ability. Results: Wood decay fungi namely, Daldinia concentrica, Ganoderma lucidum, Lenzites betulina, Ganoderma tsugae, Trametes versicolor and Schizophyllum commune, were isolated from teak wood logs as pure culture and were used for studying their degrading ability. Higher degrading percentage of 10.50 and 12.0 was recorded with L. betulina and T. versicolor inoculated wood blocks on 60th day of incubation. Similarly, the maximum moisture content of 54.45 and 50.00 percent was recorded with L. betulina and T. versicolor at 60th day of incubation in teak wood blocks. Supporting these results, L. betulina showed increased secretion of laccase, LiP and MnP enzyme activity to the level of 0.60, 0.056 and 0.26U ml-1, respectively. Interpretation: The results of the present study gives an understanding on the degrading ability of wood decay fungi which can be used to devise integrated management practices to minimise the losses under storage.

The contents of ganoderic acids in mycellium of different Ganoderma species (Ganodermataceae) obtained by different methods of cultivation

The effect of different cultivation methods on the content of ganoderic acids of 7 species, 10 strains of the Ganoderma genus (Ganoderma applanatum 1899; Ganoderma сarnosum 2502; Ganoderma lucidum 1904; Ganoderma resinaceum 2477, 2503; Ganoderma sinense 2516; Ganoderma tsugae 1848, 2024, 2566, Ganoderma oregonense 2560) genus fungi from the IBK Mushroom Culture Collection M.G. Kholodny Institute of Botany of National Academy of Sciences of Ukraine was investigated. It has been shown that the submerged cultivation method is more efficient for the accumulation of ganoderic acids for five strains. In the mycelium of the strain G. sinense 2516 was the highest content of ganoderic acids – 25.2 ± 1.5 mg / g. The productivity (yeld) of ganoderic acids synthesis is much higher with using the submerged culture cultivation method for mycelium of all used species and strains since the use of this method provides the accumulation of much more biomass in comparison with the static liquid cultivation method. The highest yield amount of ganoderic acids was in the mycelium of the G. tsugae 2024 and G. tsugae 2566 species, namely: 0.35 ± 0.019 and 0.36 ± 0.028 g / l. It was proved that the modified extraction method significantly reduces the extraction time of ganoderic acids. Extraction time is reduced from 14 to 2 days. For the G. sinense 2516 and G. tsugae 2024 strains was determined content of the ganoderic acids and their yield in dynamics of grows in the submerged culture on 6, 8, 10, 12, 14, 16, 18 and 20 day of cultivation. The highest amount of the ganoderic acids content was accumulated by the mycelium of the strain G. sinense 2516 – it was 26.4 ± 1.5 mg / g on the 14th day of cultivation. The highest yield of the ganoderic acids was in G. sinense 2516 on 14th day, and G. tsugae 2024 mycelium on the 16th day of cultivation with the next numbers 0.6 ± 0.031, 0.62 ± 0.033 and 0.62 ± 0.027 g/l.

Occurrence, diversity and morphology of poroid wood decay by Ganoderma spp. from tropical moist deciduous forest region of Bangladesh

There are many hot spots in tropical moist deciduous forest region where wood decay fungi grow naturally. An investigation was carried out to collect, identify and preserve wood decay Ganoderma spp. on the basis of morphological characteristics from Pabna, Dhaka and Rajshahi under tropical moist deciduous forest region in Bangladesh during June to September 2016 and July to October 2017. This study was conducted to record the morphological variability, distribution, habitat and diversity of the Ganoderma population. A total of 40 Ganoderma samples were collected and identified to nine species under Ganodermataceae family. The samples were collected from the sites by walking through the area following standard method. The field data and laboratory analytical data was recorded during sample collection and in the laboratory, respectively.The highest density of occurrence (23%) was recorded for Ganoderma lucidum and Ganoderma oregonense followed by Ganoderma applanatum (20%), Ganoderma praelongum (19%), Ganoderma lesklokorka (18.5%), Ganoderma pfeifferi (17%), Ganoderma boninense (15%), Ganoderma lipsiense (13%) and Ganoderma tsugae (11%). The highest frequency of occurrence (10%) was recorded for Ganoderma applanatum and Ganoderma oregonense followed by Ganoderma lucidum (9%), Ganoderma pfeifferi (8%), Ganoderma boninense (8%), Ganoderma praelongum (7%), Ganoderma lesklokorka (7%),Ganoderma tsugae (6%) and Ganoderma lipsiense (5%). During survey, Koroi (Albizia procera), Aurjun (Terminalia arjuna), Sisso plant (Dalbergia sissoo), Neem (Azadirachta indica), Golden shower (Acacia auriculiformis) and Rain tree (Albizia lebbeck) were found as hosts of Ganoderma spp. The specimens were preserved in the SAU Herbarium of Macro Fungi (SHMF) and might be useful in mushroom breeding and development program for medicine and food industry sector in future.

Ganoderma tsugae suppresses the proliferation of endometrial carcinoma cells via Akt signaling pathway.

Ganoderma is one of the common medicinal mushrooms in traditional Chinese medicine. Previous researches have unveiled the multifaceted biological activity of Ganoderma extract. Ganoderma tsugae has been investigated the potential on curing prostate, colon, lung, epidermoid, breast and ovarian cancers, but not including endometrial cancer. Endometrial cancer is a gynecological malignant tumor with serious drug resistance problem in clinical cancer treatment. This study aimed to demonstrate the first study of Ganoderma on treating endometrial cancer. The Ganoderma tsugae ethanol extract (GTEE) could suppress the proliferation of endometrial cancer cells HEC-1-A, KLE, and AN3 CA. GTEE also induced G1/S phase arrest and mitochondria-mediated apoptosis in endometrial cancer cells. Furthermore, the Akt signaling pathway could be suppressed by GTEE. Therefore, our results suggest for the first time that GTEE has the potential to be an adjuvant therapeutic agent in the treatment of endometrial cancer.

Micronized sacchachitin promotes satellite cell proliferation through TAK1-JNK-AP-1 signaling pathway predominantly by TLR2 activation

BackgroundGanoderma sp., such as Ganoderma tsugae (GT), play an important role in traditional Chinese medicine. Ganoderma sp. contains several constituents, including Sacacchin, which has recently drawn attention because it can not only enhance the repair of muscle damage but also strengthen the muscle enforcement. Although Ganoderma sp. have a therapeutic effect for neuromuscular disorders, the underlying mechanism remains unclear. This study investigated the effect and underlying molecular mechanism of micronized sacchachitin (mSC) on satellite cells (SCs), which are known as the muscle stem cells.MethodsThe myogenic cells, included SCs (Pax7+) were isolated from tibialis anterior muscles of a healthy rat and were cultured in growth media with different mSC concentrations. For the evaluation of SC proliferation, these cultivated cells were immunostained with Pax7 and bromodeoxyuridine assessed simultaneously. The molecular signal pathway was further investigated by using Western blotting and signal pathway inhibitors.ResultsOur data revealed that 200 µg/mL mSC had an optimal capability to significantly enhance the SC proliferation. Furthermore, this enhancement of SC proliferation was verified to be involved with activation of TAK1-JNK-AP-1 signaling pathway through TLR2, whose expression on SC surface was confirmed for the first time here.ConclusionMicronized sacchachitin extracted from GT was capable of promoting the proliferation of SC under a correct concentration.

Preparation of Nano/Submicron Ganoderma Tsugae and Its Stability

The preparation and stability of nano/submicrometer-sized Ganoderma tsugae (G. tsugae) was explored. The fruit body of G. tsugae was treated by homogenization and media-milling. Three sizes (0.8, 0.3, and 0.2 mm) of media were employed for consecutive milling. The final product was obtained by centrifugation (10,000 g, 10 min). Particle sizes were determined by using a laser-light particle size analyzer and further examined by using transmission electron microscopy. The removal of large particles resulted in better stability. After being stored at25°C for 21 days, the volume-average diameter of supernatant III increased from 0.105 to 0.137 μm. Freeze-drying resulted in particle aggregation and an increase in particle size. Twenty percent of the particles in the freeze-dried supernatant III were smaller than 100 nm, with an average diameter greater than 1 μm. Nevertheless, autoclaving did not result in severe instability of supernatant III. After being stored at 25°C for 12 months, about 40% of the particles were smaller than 100 nm, with an average diameter of 373 nm. In addition, the contents of bioactive compounds such as β-(1,3)-D-glucans, crude triterpenoids, total dietary fiber and chitin in media-milled products were much greater than those present in the hot-water extract.

Determination of ganoderic acids in triterpenoid constituents of Ganoderma tsugae

Ganoderma product has the highest sale volume in the health food market in Taiwan. The most widely-used raw material is the species of G. tsuage. Nine triterpenoids were isolated from the fruiting body of G. tsugae and their structures were identified, using 1H NMR and FT-IR, as ganoderic acid A (1), B (2), C (3), D (4), E (5), C5 (6), C6 (7), G (8) and ganoderenic acid D (9). A reverse-phase HPLC method was also established for the determination of these nine triterpenoids. Among these compounds, ganoderic acid E, C5, C6, G and ganoderenic acid D were analyzed for the first time by reverse-phase HPLC. The separation was carried out on a C18 reverse-phase column with a gradient elution of acetonitrile and 2% acetic acid (1/4 and 1/2, v/v) as mobile phase at a flow-rate of 0.8 mL/min and detection at 252 nm. Regression equation revealed the linear relationship (correlation coefficient: 0.9990-0.9999) between the peak-area ratios and concentrations of ganoderic acids. The recoveries of nine ganoderic acids were between 96.85-105.09%. The relative standard deviations of nine ganoderic acids range from 0.8-4.8% (intraday) and 0.7-5.1% (interday). The total contents of the nine ganoderic acids in eight Ganoderma samples were between 0.28-2.20%.

Antioxidant Activities of Selected Medicinal Mushroom Submerged Cultivated Mycelia.

Medicinal mushrooms fruiting bodies have been used as food or medicine for years but cultured mycelium is faster to grow and costs less. This research studied the antioxidant activities of three species (five strains) of medicinal mushroom mycelia (Cordyceps militaris, Ganoderma tsugae I and II, Trametes versicolor I and II). Two-stage extractions were performed: first the sample was extracted with 70% ethanol, and then the residue was extracted with 95°C hot water. Both ethanolic and hot water extracts showed effective concentration (EC50) values of 0.29-4.22 mg/mL, indicating that these extracts were remarkably effective in antioxidant activities. The ethanolic extracts displayed more effective reducing power, scavenging, and chelating ability (EC50 0.33-2.37 mg/mL) than hot water extracts (EC50 0.58-4.22 mg/g). Besides, ethanolic extracts contained higher total phenol content (75.49-144.99 GAE mg/g) than the hot water extracts (22.77-58.68 GAE mg/g). Furthermore, the ethanolic extracts contained flavonoids but not the hot water extract. Overall, these mycelia were highly effective in the antioxidant activities and might be potent antioxidants.

Effect of Shading and Forest Type on Morphological Characteristics and Bioactive Compounds of Fruiting Bodies of Songshan Lingzhi Ganoderma tsugae (Agaricomycetes).

Ganoderma tsugae strain MCCCMAS0053 cultivation on short logs results in varied yield and quality under different growth conditions. Thus, growth conditions need optimization to increase yield and quality. An indoor experiment with three shade treatments (A1, two layers of black sun-shade net; A2, one layer of black sun-shade net plus plastic mulch; A3, one layer of black sun-shade net) and a field experiment in two forest types (pine or mixed pine-oak) were conducted. The results showed that shading and forest type significantly affected light intensities and the growth, size, biomass, and bioactive components of G. tsugae fruiting bodies. In the indoor experiment, the mean dry weight of the fruiting body and the diameter of the pileus in A2 increased by 21.51-44.98% and 13.42-22.26%, respectively, compared with those of A1 and A3. Similarly, the accumulation of the bioactive compounds (polysaccharides, total amino acids, and total essential amino acids) in the pileus and stipe were greater in A2 than in A1 and A3. Furthermore, compared with pure pine forest cultivation, fruiting bodies cultivated in mixed pine-oak forest had greater dry weight, pileus diameter, and more bioactive compounds. In addition, no significant difference was found between the A2 and mixed pine-oak treatments, which had a similar light intensity (from 1116 to 2367 lx). Hence, this suggests that the A2 shade treatment or cultivation in mixed pine-oak forest is beneficial for production of G. tsugae fruiting bodies, and light intensity may play a critical role in this process.

Wasted Ganoderma tsugae Derived Chitosans for Smear Layer Removal in Endodontic Treatment

The objective of this study is to investigate the synergistic effects of acid etching and metal-ion chelation in dental smear layer removal using wasted Ganoderma tsugae derived chitosans. The wasted Ganoderma tsugae fruiting body was used to prepare both acid-soluble fungal chitosan (FCS) and alkali-soluble polysaccharide (ASP). To explore the effective irrigant concentration for smear layer removal, a chelating effect on ferrous ions was conducted. Specimens of various concentrations of EDTA, citric acid, and polysaccharide solutions were reacted with FerroZine™ then the absorbance was examined at 562 nm by a UV-visible spectrophotometer to calculate their metal chelating capability. Twenty extracted premolars were instrumented and individually soaked in the solutions of 15 wt% EDTA, 10 wt% citric acid, 0.04 wt% ASP, 0.04 wt% FCS, and normal saline were randomly divided into five groups (N=4). Next, each tooth was cleaved longitudinally and examined by scanning electron microscopy (SEM) to assay the effectiveness of smear layer removal. The chelating capability for EDTA, FCS, and ASP showed no significant difference over the concentration of 0.04 wt% (p > 0.05). The SEM results showed that 0.04 wt% FCS solution was effective in smear layer removal along the canal wall. These results indicated that Ganoderma tsuage derived FCS in acid solutions could be a potential alternative as a root canal irrigant solution due to its synergistic effect.