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  1. A novel study based on adaptive metal tolerance behavior in fungi and SEM-EDX analysis
    Chen SH, Ng SL, Cheow YL, Ting ASY
    J Hazard Mater, 2017 Jul 15;334:132-141.
    PMID: 28407540 DOI: 10.1016/j.jhazmat.2017.04.004
    Four fungal isolates: Simplicillium chinense (iso 9, accession no. KX425621), Penicillium simplicissimum (iso 10, KP713758), Trichoderma asperellum (iso 11, KP792512), and Coriolopsis sp. (1c3, KM403574) were subjected to a series of induced-tolerance training under high metal concentrations to determine if greater tolerance could be achieved from constant exposure to such conditions. Adaptive tolerance assay (Tolerance Index, TI) and Field-Emission Scanning Electron Microscopy with Energy Dispersive X-ray (SEM-EDX) characterized their metal tolerance. "Untrained" S. chinense, P. simplicissimum and T. asperellum showed tolerance towards 4000-4500ppm Al(III) (TI: 0.64-0.71), 1000ppm Cr(III) (0.52-0.83) and Pb(II) (0.32-0.88). With tolerance training, tolerance towards 2000-6000ppm Al(III), 500-3000ppm Pb(II) and 2000-3000ppm Cr(III) were achieved (TI: 0.01-0.82) compared to untrained cultures (0.00-0.59). In contrast, tolerance training for Coriolopsis sp. and P. simplicissimum was less successful, with TI values similar or lower than untrained cultures. SEM-EDX analysis proposed biosorption and bioaccumulation as mechanisms for metal removal. The latter was demonstrated with the removal of Cr(III) and Pb(II) by S. chinense (12.37 and 11.52mgg-1, respectively) and T. asperellum (10.44 and 7.50mgg-1). Induced-tolerance training may render benefit in the long run, but this delicate approach is suggestively species and metal dependent.
    Matched MeSH terms: Polyporaceae/drug effects*; Polyporaceae/physiology; Polyporaceae/ultrastructure
  2. Mutagenicity and genotoxicity effects of Lignosus rhinocerotis mushroom mycelium
    Chen TI, Zhuang HW, Chiao YC, Chen CC
    J Ethnopharmacol, 2013 Aug 26;149(1):70-4.
    PMID: 23773827 DOI: 10.1016/j.jep.2013.06.001
    Lignosus rhinocerotis mushroom is widely used as traditional medicine and as soup ingredient in Malaysia and Hong Kong. Its sclerotium is the part of edibility and is traditionally used for the treatment of fever, cough, asthma and cancer. In view of its safety profile, very little information is found in scientific literature.
    Matched MeSH terms: Polyporaceae/chemistry*
  3. Cloning, overexpression, purification, and modeling of a lectin (Rhinocelectin) with antiproliferative activity from Tiger Milk Mushroom, Lignosus rhinocerus
    Cheong PCH, Yong YS, Fatima A, Ng ST, Tan CS, Kong BH, et al.
    IUBMB Life, 2019 10;71(10):1579-1594.
    PMID: 31190445 DOI: 10.1002/iub.2101
    A lectin gene from the Tiger Milk Mushroom Lignosus rhinocerus TM02® was successfully cloned and expressed via vector pET28a in Escherichia coli BL21(DE3). The recombinant lectin, Rhinocelectin, with a predicted molecular mass of 22.8 kDa, was overexpressed in water-soluble form without signal peptide and purified via native affinity chromatography Ni-NTA agarose. Blast protein analysis indicated the lectin to be homologous to jacalin-related plant lectin. In its native form, Rhinocelectin exists as a homo-tetramer predicted with four chains of identical proteins consisting of 11 beta-sheet structures with only one alpha-helix structure. The antiproliferative activity of the Rhinocelectin against human cancer cell lines was concentration dependent and selective. The IC50 values against triple negative breast cancer cell lines MDA-MB-231 and breast cancer MCF-7 are 36.52 ± 13.55 μg mL-1 and 53.11 ± 22.30 μg mL-1 , respectively. Rhinocelectin is only mildly cytotoxic against the corresponding human nontumorigenic breast cell line 184B5 with IC50 value at 142.19 ± 36.34 μg mL-1 . The IC50 against human lung cancer cell line A549 cells is 46.14 ± 7.42 μg mL-1 while against nontumorigenic lung cell line NL20 is 41.33 ± 7.43 μg mL-1 . The standard anticancer drug, Doxorubicin exhibited IC50 values mostly below 1 μg mL-1 for the cell lines tested. Flow cytometry analysis showed the treated breast cancer cells were arrested at G0/G1 phase and apoptosis induced. Rhinocelectin agglutinated rat and rabbit erythrocytes at a minimal concentration of 3.125 μg mL-1 and 6.250 μg mL-1 , respectively.
    Matched MeSH terms: Polyporaceae/genetics*; Polyporaceae/chemistry
  4. Differential identification of mushrooms sclerotia by IR macro-fingerprint method
    Choong YK, Lan J, Lee HL, Chen XD, Wang XG, Yang YP
    Spectrochim Acta A Mol Biomol Spectrosc, 2016 Jan 5;152:34-42.
    PMID: 26186395 DOI: 10.1016/j.saa.2015.07.054
    Many macrofungus sclerotia are well-known medicinal herbs, health food and nutritional supplements. However, the prevalent adulterant commercial products are major hindrances to their incorporation into mainstream medical use in many countries. The mushroom sclerotia of Lignosus rhinocerotis, Poria cocos, Polyporus umbellatus, Pleurotus tuber-regium and Omphalia lapidescens are commonly used in traditional Chinese medicine. In this study, IR macro-fingerprint method was used in the identification of these sclerotia. The results showed that the spectrum of L. rhinocerotis (LR) was comparable with P. cocos with 94.4% correlation, except that the peak at 1543cm(-1) of LR appeared in lower intensity. The spectrum of P. umbellatus and P. tuber-regium was also correlated (91.5%), as both spectra could be clearly discriminated in that P. umbellatus spectrum has small base peaks located at the range of 1680-1500cm(-1). O. lapidescens was not comparable with all the other sclerotia as its spectrum was totally different. Its base peak was broad and derivated equally along the range. The first IR has revealed the dissimilarity among five mushrooms sclerotia. The second derivative and 2DIR further enhanced the identification in detail.
    Matched MeSH terms: Polyporaceae
  5. Fulltext Lignosus rhinocerus (Cooke) Ryvarden: A Medicinal Mushroom That Stimulates Neurite Outgrowth in PC-12 Cells
    Eik LF, Naidu M, David P, Wong KH, Tan YS, Sabaratnam V
    Evid Based Complement Alternat Med, 2012;2012:320308.
    PMID: 22203867 DOI: 10.1155/2012/320308
    A national treasure mushroom, Lignosus rhinocerus, has been used to treat variety of ailments by local and indigenous communities in Malaysia. The aim of this study was to investigate the potential of the most valuable part of L. rhinocerus, the sclerotium, on neurite outgrowth activity by using PC-12Adh cell line. Differentiated cells with one thin extension at least double the length of the cell diameter were scored positive. Our results showed that aqueous sclerotium L. rhinocerus extract induced neurite outgrowths of 24.4% and 42.1% at 20 μg/mL (w/v) of aqueous extract alone and a combination of 20 μg/mL (w/v) aqueous extract and 30 ng/mL (w/v) of NGF, respectively. Combination of NGF and sclerotium extract had additive effects and enhanced neurite outgrowth. Neuronal differentiation was demonstrated by indirect immunofluorescence of neurofilament protein. Aqueous sclerotium extract contained neuroactive compounds that stimulated neurite outgrowth in vitro. To our knowledge this is the first report on neurite-stimulating activities of L. rhinocerus.
    Matched MeSH terms: Polyporaceae
  6. Functional evaluation of a recombinant fungal immunomodulatory protein from L. rhinocerus produced in P. pastoris and E. coli host expression systems
    Ejike UC, Chan CJ, Lim CSY, Lim RLH
    Appl Microbiol Biotechnol, 2021 Apr;105(7):2799-2813.
    PMID: 33763709 DOI: 10.1007/s00253-021-11225-x
    Fungal immunomodulatory proteins (FIPs) are bioactive proteins with immunomodulatory properties. We previously reported the heterologous production in Escherichia coli of FIP-Lrh from Tiger milk mushroom (Lignosus rhinocerus) with potent cytotoxic effect on cancer cell lines. However, protein produced in E. coli lacks post-translational modifications and may be contaminated with lipopolysaccharide (LPS) endotoxin. Therefore, in this study, yFIP-Lrh produced in Pichia pastoris was functionally compared with eFIP-Lrh produced in E. coli. Expression construct of FIP-Lrh cDNA in pPICZα was generated, transformed into P. pastoris X-33 and Mut+ transformants were verified by colony PCR. Induction with 0.5% or 1% methanol resulted in a secreted 13.6 kDa yFIP-Lrh which was subsequently purified and verified using LCMS/MS analysis. Size exclusion chromatography confirmed eFIP-Lrh as a homodimer whereas the larger size of yFIP-Lrh may indicate post-translational modification despite negative for glycoproteins staining. At lower concentration (4-8 μg/mL), yFIP-Lrh induced significantly higher Th1 (IFN-γ, TNF-α) and Th2 (IL-6, IL-4, IL-5, IL-13) cytokines production in mice splenocytes, whereas 16 μg/mL eFIP-Lrh induced significantly higher pro-inflammatory cytokines (TNF-α, IL-6, IL-10), possibly due to higher residual LPS endotoxin (0.082 EU/mL) in eFIP-Lrh compared to negligible level in yFIP-Lrh (0.001 EU/mL). Furthermore, yFIP-Lrh showed higher cytotoxic effect on MCF-7 and HeLa cancer cells. Since both recombinant proteins of FIP-Lrh have the same peptide sequence, besides glycosylation, other post-translational modifications in yFIP-Lrh may account for its enhanced immunomodulatory and anti-proliferative activities. In conclusion, P. pastoris is preferred over E. coli for production of a functionally active yFIP-Lrh devoid of endotoxin contamination. KEY POINTS: • FIP-Lrh can induced production of Th1 and Th2 cytokines by mouse splenocytes. • Higher cytotoxic effect on cancer cells observed for yeast compared to E. coli produced FIP-Lrh. • P. pastoris allows production of an endotoxin-free and functionally active recombinant FIP-Lrh.
    Matched MeSH terms: Polyporaceae
  7. Nutritional evaluation on Lignosus cameronensis C. S. Tan, a medicinal Polyporaceae
    Fung SY, Cheong PCH, Tan NH, Ng ST, Tan CS
    IUBMB Life, 2019 07;71(7):821-826.
    PMID: 30629799 DOI: 10.1002/iub.2006
    Sclerotial powder of a cultivated species of the Tiger Milk Mushroom, Lignosus cameronensis was analysed for its nutritional components and compared against species of the same genus, Lignosus rhinocerus and Lignosus tigris. All three species have been used by indigenous tribes in Peninsular Malaysia as medicinal mushrooms. Content of carbohydrate, fibre, mineral, amino acid, palatable index, fat, ash and moisture were determined. L. cameronensis sclerotial material consists of carbohydrate (79.7%), protein (12.4%) and dietary fibre (5.4%) with low fat (1.7%) and no free sugar. It has the highest content of total carbohydrate (791 g kg-1 ), energy value (3,700 kcal kg-1 ) and calcium (0.85 g kg-1 ). The crude protein content (123 g kg-1 ) is comparable to that of L. rhinocerus with its main amino acids consisting of glutamic acid, aspartic acid and leucine. The umami index is determined to be 0.27. The total essential amino acid (45 g kg-1 ) is comparable to that of L. tigris. The main mineral is potassium (1.51 g kg-1 ) and the Na/K ratio was <0.6. Heavy metals such as mercury, cadmium, lead and arsenic were absent. L. cameronensis has the highest amount of food energy, total carbohydrate and calcium compared to those of both L. rhinocerus and L. tigris. The essential amino acids comprised almost 40% of the total amino acid content, slightly more than that reported from sclerotial powder of the L. tigris. © 2019 IUBMB Life, 9999(9999):1-6, 2019.
    Matched MeSH terms: Polyporaceae/growth & development; Polyporaceae/metabolism*
  8. Impact of electrical stimulation on the growth of mycelium of lignosus rhinocerus (cooke) ryvarden
    Jamil NAM, Gomes C, Kadir Z, Gomes A
    Electromagn Biol Med, 2020 Oct 01;39(4):356-363.
    PMID: 32878496 DOI: 10.1080/15368378.2020.1812080
    Corona discharge from multiple needles at an electrical potential of 5 kV generated by a Van de Graff generator increased the growth rate of the mycelium of tiger's milk mushroom by 10.3% at the end of the first eight days. A similar growth rate enhancement was observed for the next eight days as well. Mycelium of tiger's milk mushroom was cultured on agar media in Petri dish for five days prior to the exposure to various forms of electrical stimulations. The direct current injection (1.1-1.3 A) to the growing medium, application of an electric potential to the growing environment at low strength (30 V) and high strength (5 kV) with single and multiple needles showed varying degrees of success. This suggests that the mycelium of tiger's milk mushroom could positively be stimulated by specific electrical stimulation techniques with selected parameters. This will pave the way to a highly beneficial growth enhancement technique that can be up-scaled to apply in mass production of mushroom.
    Matched MeSH terms: Polyporaceae/growth & development*
  9. Comparative nutritional and mycochemical contents, biological activities and LC/MS screening of tuber from new recipe cultivation technique with wild type tuber of tiger's milk mushroom of species Lignosus rhinocerus
    Jamil NAM, Rashid NMN, Hamid MHA, Rahmad N, Al-Obaidi JR
    World J Microbiol Biotechnol, 2017 Dec 04;34(1):1.
    PMID: 29204733 DOI: 10.1007/s11274-017-2385-4
    Tiger's milk mushroom is known for its valuable medicinal properties, especially the tuber part. However, wild tuber is very hard to obtain as it grows underground. This study first aimed to cultivate tiger's milk mushroom tuber through a cultivation technique, and second to compare nutritional and mycochemical contents, antioxidant and cytotoxic activities and compound screening of the cultivated tuber with the wild tuber. Results showed an increase in carbohydrate content by 45.81% and protein content by 123.68% in the cultivated tuber while fat content reduced by 13.04%. Cultivated tuber also showed an increase of up to 64.21% for total flavonoid-like compounds and 62.51% of total β-D-glucan compared to the wild tuber. The antioxidant activity of cultivated tuber and wild tuber was 760 and 840 µg mL-1, respectively. The cytotoxic activity of boiled water extract of cultivated tuber against a human lung cancer cell line (A549) was 65.50 ± 2.12 µg mL-1 and against a human breast cancer cell line (MCF7) was 19.35 ± 0.11 µg mL-1. β-D-glucan extract from the purification of boiled water extract of cultivated tuber showed cytotoxic activity at 57.78 ± 2.29 µg mL-1 against A549 and 33.50 ± 1.41 µg mL-1 against MCF7. However, the β-glucan extract from wild tuber did not show a cytotoxic effect against either the A549 or MCF7 cell lines. Also, neither of the extracts from cultivated tuber and wild tuber showed an effect against a normal cell line (MRC5). Compound profiling through by liquid chromatography mass spectrometry (LC/MS) showed the appearance of new compounds in the cultivated tuber. In conclusion, our cultivated tuber of tiger's milk mushroom using a new recipe cultivation technique showed improved nutrient and bioactive compound contents, and antioxidant and cytotoxic activities compared to the wild tuber. Further investigations are required to obtain a better quality of cultivated tuber.
    Matched MeSH terms: Polyporaceae/chemistry*
  10. Oral reproductive and developmental toxicity of Lignosus rhinocerotis mycelium in rat
    Jhou BY, Liu HH, Yeh SH, Chen CC
    J Ethnopharmacol, 2017 Aug 17;208:66-71.
    PMID: 28663148 DOI: 10.1016/j.jep.2017.06.029
    ETHNOPHARMACOLOGICAL RELEVANCE: Lignosus rhinocerotis (L. rhinocerotis), also known as the tiger milk mushroom, is widely used as traditional medicine and as soup ingredient in Malaysia and Hong Kong. It is edible and is used traditionally for the treatment of fever, cough, asthma, wounds, chronic hepatitis, gastric ulcers and cancers. In view of its safety profile, little information is found in scientific literature. The objective of this study is to investigate developmental toxicity of L. rhinocerotis in pregnant Sprague-Dawley (SD) rats.

    MATERIALS AND METHODS: Eighty pregnant SD female rats were used in this study for three treatment groups and a control group, each consisting of 20 pregnant female rats. Three doses of 850mg/kg/day (Low-dose), 1700mg/kg/day (Mid-dose) and 3400mg/kg/day (High-dose) were selected for the study, whereas 10mL/kg distilled water was served as the control. Examinations were conducted on pregnant rats and fetuses respects to mortality, body weight, body weights gains, food consumption and clinical observations. The pregnant females were gross necropsied on G20, followed by maternal and fetus examination, to evaluate the teratogenicity, reproductive and developmental performance of L. rhinocerotis mycelium.

    RESULTS: Results showed that no L. rhinocerotis mycelium-related animal death and abnormal clinical sign were noted. No statistical differences were noted in maternal mean body weight and maternal mean body weight gains. Some animals in the high-dose group appeared audible respiration due to dosing accident, it resulted in lower food consumption but not relevant to L. rhinocerotis mycelium treatment. In maternal gross necropsy, no L. rhinocerotis mycelium-related gross lesion was noted. In maternal examination, parameters of gravid uterus weight, implantation number, corpora lutea number, litter size, live or dead fetal number, male or female fetus number, resorption number, fetal sex ratio (M/F), pre-implantation loss and post-implantation loss were all within the normal reference ranges and showed no significant difference when compared to the control group. In fetus examination, including external, visceral and skeletal evaluations, there were no significant changes between any of the L. rhinocerotis mycelium treated groups and the control group.

    CONCLUSIONS: Based on the study results, the no-observable-adverse-effect level (NOAEL) for pregnant female rats under the conditions of this study was 3400mg/kg/day.

    Matched MeSH terms: Polyporaceae*
  11. Fractionation and Biological Activities of Water-Soluble Polysaccharides from Sclerotium of Tiger Milk Medicinal Mushroom, Lignosus rhinocerotis (Agaricomycetes)
    Keong CY, B V, Daker M, Hamzah MY, Mohamad SA, Lan J, et al.
    Int J Med Mushrooms, 2016;18(2):141-54.
    PMID: 27279536 DOI: 10.1615/IntJMedMushrooms.v18.i2.50
    This study investigated antioxidant and anti-inflammatory properties, and the direct cytotoxic effect of Lignosus rhinocerotis fractions, especially the polysaccharide fraction, on nasopharyngeal carcinoma cells. L. rhinocerotis crude extract was obtained through hot water extraction. The precipitate saturated with 30% ammonium sulfate was purified with ion-exchanged chromatography. Gel permeation chromatography multiangle laser light scattering analysis equipped with light scattering and UV signals revealed two district groups of polymers. A total of four peaks were observed in the total carbohydrate test. Fraction C, which was the second region of the second peak eluted with 0.3 M NaOH, showed the highest integrated molecular weight, whereas fraction E had the lowest integrated molecular weight of 19,790 Da. Fraction A contained the highest β-D-glucan content. Enzymatic analysis showed that most of the polysaccharide fractions contained β-1-3 and β-1-6 skeletal backbones. The peak eluted with 0.6 M NaOH was separated in fraction D (flask 89-92) and fraction E (93-96). The results showed that fraction E expressed higher antioxidant activities than fraction D whereas fraction D expressed higher chelating activity than fraction E. The extract saturated with 30% ammonium sulfate exhibited higher reducing power than the extract saturated with 100% ammonium sulfate. Fractions D and E significantly inhibited the secretion of tumor necrosis factor-α in lipopolysaccharide-stimulated RAW 264.7 macrophages in a dose-dependent manner. There was no apparent difference in the viability of cells exposed or unexposed to L. rhinocerotis fractions.
    Matched MeSH terms: Polyporaceae/chemistry*
  12. Nutritional composition, antioxidant properties, and toxicology evaluation of the sclerotium of Tiger Milk Mushroom Lignosus tigris cultivar E
    Kong BH, Tan NH, Fung SY, Pailoor J, Tan CS, Ng ST
    Nutr Res, 2016 Feb;36(2):174-83.
    PMID: 26598045 DOI: 10.1016/j.nutres.2015.10.004
    The Tiger Milk Mushroom (Lignosus spp.) is an important medicinal mushroom in Southeast Asia and has been consumed frequently by the natives as a cure for a variety of illnesses. In this study, we hypothesized that Lignosus tigris (cultivar E) sclerotium may contain high nutritional value and antioxidant properties, is nontoxic and a potential candidate as a dietary supplement. The chemical and amino acid compositions of the sclerotium were evaluated and antioxidant activities of the sclerotial extracts were assessed using ferric reducing antioxidant power; 1,1-diphenyl-2-picrylhydrazyl; and superoxide anion radical scavenging assays. Acute toxicity of the L. tigris E sclerotium was assessed using a rat model study. The sclerotium was found to be rich in carbohydrate, protein, and dietary fibers with small amounts of fat, calories, and sugar. The amino acid composition of the protein contains all essential amino acids, with a protein score of 47. The sclerotial extracts contain phenolics, terpenoids, and glucan. The ferric reducing antioxidant power values of the various sclerotial extracts (hot water, cold water, and methanol) ranged from 0.008 to 0.015 mmol min(-1) g(-1) extract, while the 1,1-diphenyl-2-picrylhydrazyl and superoxide anion radical scavenging activities ranged from 0.11 to 0.13, and -2.81 to 9.613 mmol Trolox equivalents g(-1) extract, respectively. Acute toxicity assessment indicated that L. tigris E sclerotial powder was not toxic at the dose of 2000 mg kg(-1). In conclusion, L. tigris E sclerotium has the potential to be developed into a functional food and nutraceutical.
    Matched MeSH terms: Polyporaceae/chemistry*
  13. Fulltext Sub-Acute Toxicity Study of Tiger Milk Mushroom Lignosus tigris Chon S. Tan Cultivar E Sclerotium in Sprague Dawley Rats
    Kong BH, Tan NH, Fung SY, Pailoor J
    Front Pharmacol, 2016;7:246.
    PMID: 27555822 DOI: 10.3389/fphar.2016.00246
    Lignosus also known as "Tiger Milk Mushroom," is classified in the family Polyporaceae and mainly consumed for its medicinal properties in Southeast Asia and China. The sclerotium is known as the part with medicinal value and often used by the natives to treat a variety of ailments. Lignosus tigris Chon S. Tan, one of the species of the Malaysia Tiger Milk mushroom, has recently been successfully cultivated in laboratory. Earlier studies have demonstrated the L. tigris cultivar E sclerotia exhibited beneficial biomedicinal properties. This study evaluated the potential toxicity of L. tigris E sclerotia in a 28-day sub-acute oral administration in Sprague Dawley (SD) rats. L. tigris E sclerotial powder was administered orally at three different doses of 250, 500, and 1000 mg/kg to the SD rats once daily, consecutively for 28-days. Body weight of the rats was recorded and general behavior, adverse effects, and mortality were observed daily throughout the experimental period. At the end of the experiment, blood hematology and biochemistry, relative organ weights, and histopathological analysis were performed. Results showed that there were no mortality nor signs of toxicity throughout the 28-day sub-acute toxicity study. Oral administration of the L. tigris E sclerotial powder at daily dose up to 1000 mg/kg had no significant effects in body weight, relative organ weight, blood hematological and biochemistry, gross pathology, and histopathology of the organs. L. tigris E sclerotial powder did not cause any treatment-related adverse effect in the rats at different treatment dosages up to 1000 mg/kg. As the lethal dose for the rats is above 1000 mg/kg, the no-observed-adverse-effect level (NOAEL) dose is more than 1000 mg/kg.
    Matched MeSH terms: Polyporaceae
  14. Longitudinal Assessment of the Tiger Milk Medicinal Mushroom, Lignosus rhinocerus (Agaricomycetes), Sclerotium Cultivar: Uniformity of Bioactive Components
    Kong BH, Fung SY
    Int J Med Mushrooms, 2021;23(10):61-68.
    PMID: 34595892 DOI: 10.1615/IntJMedMushrooms.2021040120
    Traditional use of the tiger milk medicinal mushroom, Lignosus rhinocerus, to treat various illnesses has been recorded for > 4 centuries. Successful cultivation of L. rhinocerus using proprietary solid-state fermentation (SSF) technology by LiGNO Biotech has enabled large-scale production of L. rhinocerus sclerotia (termed L. rhinocerus TM02) and further investigations into its medicinal properties. Pharmacological activities of L. rhinocerus TM02, including its antioxidant, anti-inflammatory, anticancer, and immunomodulatory effects and the bioactive components responsible, have been validated by various scientific studies. In this study, we assessed the consistency of the bioactive components in 11 batches of L. rhinocerus TM02 produced over a 9-year period. The different batches of L. rhinocerus TM02 consisted of stable protein, polysaccharide, and glycoprotein contents, and all tested samples were comparable to the wild type. L. rhinocerus TM02 had greater protein, carbohydrate, and glycoprotein contents, which were mostly bioactive compared to another cultivar from a different cultivation technology (TM-UN). Together with previous scientific validations, L. rhinocerus TM02 produced using SSF cultivation is of optimal quality with high consistent bioactive contents, which can be an appropriate indicator for quality validation of the much sought-after medicinal mushroom, L. rhinocerus.
    Matched MeSH terms: Polyporaceae*
  15. Fulltext The potential of mycelium and culture broth of Lignosus rhinocerotis as substitutes for the naturally occurring sclerotium with regard to antioxidant capacity, cytotoxic effect, and low-molecular-weight chemical constituents
    Lau BF, Abdullah N, Aminudin N, Lee HB, Yap KC, Sabaratnam V
    PLoS One, 2014;9(7):e102509.
    PMID: 25054862 DOI: 10.1371/journal.pone.0102509
    Previous studies on the nutritional and nutraceutical properties of Lignosus rhinocerotis focused mainly on the sclerotium; however, the supply of wild sclerotium is limited. In this investigation, the antioxidant capacity and cytotoxic effect of L. rhinocerotis cultured under different conditions of liquid fermentation (shaken and static) were compared to the sclerotium produced by solid-substrate fermentation. Aqueous methanol extracts of the mycelium (LR-MH, LR-MT) and culture broth (LR-BH, LR-BT) demonstrated either higher or comparable antioxidant capacities to the sclerotium extract (LR-SC) based on their radical scavenging abilities, reducing properties, metal chelating activities, and inhibitory effects on lipid peroxidation. All extracts exerted low cytotoxicity (IC50>200 µg/ml, 72 h) against selected mammalian cell lines. Several low-molecular-weight compounds, including sugars, fatty acids, methyl esters, sterols, amides, amino acids, phenolics, and triterpenoids, were identified using GC-MS and UHPLC-ESI-MS/MS. The presence of proteins (<40 kDa) in the extracts was confirmed by SDS-PAGE and SELDI-TOF-MS. Principal component analysis revealed that the chemical profiles of the mycelial extracts under shaken and static conditions were distinct from those of the sclerotium. Results from bioactivity evaluation and chemical profiling showed that L. rhinocerotis from liquid fermentation merits consideration as an alternative source of functional ingredients and potential substitute for the sclerotium.
    Matched MeSH terms: Polyporaceae/growth & development; Polyporaceae/chemistry*
  16. Chemical composition and cellular toxicity of ethnobotanical-based hot and cold aqueous preparations of the tiger's milk mushroom (Lignosus rhinocerotis)
    Lau BF, Abdullah N, Aminudin N, Lee HB
    J Ethnopharmacol, 2013 Oct 28;150(1):252-62.
    PMID: 23993912 DOI: 10.1016/j.jep.2013.08.034
    The sclerotium of the "tiger's milk mushroom" (Lignosus rhinocerotis) is used as tonic and folk medicine for the treatment of cancer, fever, cough and asthma by the local and indigenous communities. It is traditionally prepared by either boiling or maceration-like methods; however, there is no attempt to understand how different processing methods might affect their efficacies as anticancer agents.
    Matched MeSH terms: Polyporaceae*
  17. Chemical composition of the tiger's milk mushroom, Lignosus rhinocerotis (Cooke) Ryvarden, from different developmental stages
    Lau BF, Abdullah N, Aminudin N
    J Agric Food Chem, 2013 May 22;61(20):4890-7.
    PMID: 23597270 DOI: 10.1021/jf4002507
    The chemical composition of the tiger's milk mushroom (Lignosus rhinocerotis) from different developmental stages, i.e., the fruit body, sclerotium, and mycelium, was investigated for the first time. The fruit body and sclerotium of L. rhinocerotis were rich in carbohydrates and dietary fibers but low in fat. Protein levels in L. rhinocerotis were moderate, and all essential amino acids, except tryptophan, were present. The mycelium contained high levels of potassium, phosphorus, magnesium, riboflavin, and niacin and appreciable amounts of essential fatty acids. The results indicated that the sclerotium of L. rhinocerotis that was used in ethnomedicine was not superior to the fruit body and mycelium with regard to the nutritional content and bioactive constituents. Our findings provide some insights into the selection of appropriate mushroom part(s) of L. rhinocerotis and proper cultivation techniques for the development of new nutraceuticals or dietary supplements.
    Matched MeSH terms: Polyporaceae/growth & development*; Polyporaceae/chemistry*
  18. Comparative SELDI-TOF-MS profiling of low-molecular-mass proteins from Lignosus rhinocerus (Cooke) Ryvarden grown under stirred and static conditions of liquid fermentation
    Lau BF, Aminudin N, Abdullah N
    J Microbiol Methods, 2011 Oct;87(1):56-63.
    PMID: 21801760 DOI: 10.1016/j.mimet.2011.07.005
    Mushrooms are considered as important source of biologically active compounds which include low-molecular-mass protein/peptides (LMMP). In this study, we attempted to profile the LMMP from Lignosus rhinocerus, a wild medicinal mushroom, grown by static cultures (SC) and in stirred tank reactor (STR). Crude water extract (CWE) and protein fractions were profiled using H50 ProteinChip® arrays and SELDI-TOF-MS. Three protein peaks of 5.8, 6.9 and 9.1 kDa were found to be common to spectra of L. rhinocerus CWE from both culture conditions. Partial protein purification has resulted in detection of more peaks in the spectra of protein fractions. For protein fractions of L. rhinocerus cultured in STR, most peaks were observed in the range of 3-8 kDa whereas some peaks with molecular mass up to 14.3 kDa were noted in spectra of protein fractions from SC. Our results have demonstrated the optimization of profiling method using SELDI-TOF-MS for fungal LMMP.
    Matched MeSH terms: Polyporaceae/growth & development; Polyporaceae/metabolism*; Polyporaceae/chemistry
  19. Ethnomedicinal uses, pharmacological activities, and cultivation of Lignosus spp. (tiger׳s milk mushrooms) in Malaysia - A review
    Lau BF, Abdullah N, Aminudin N, Lee HB, Tan PJ
    J Ethnopharmacol, 2015 Jul 1;169:441-58.
    PMID: 25937256 DOI: 10.1016/j.jep.2015.04.042
    Several members of the genus Lignosus, which are collectively known as cendawan susu rimau (in Malay) or tiger׳s milk mushrooms (TMM), are regarded as important local medicine particularly by the indigenous communities in Malaysia. The mushroom sclerotia are purportedly effective in treating cancer, coughs, asthma, fever, and other ailments. The most commonly encountered Lignosus spp. in Malaysia was authenticated as Lignosus rhinocerotis (Cooke) Ryvarden (synonym: Polyporus rhinocerus), which is also known as hurulingzhi in China and has been used by Chinese physicians to treat liver cancer, gastric ulcers, and chronic hepatitis. In spite of growing interest in the therapeutic potential of TMM, there is no compilation of scientific evidence that supports the ethnomedicinal uses of these mushrooms. Therefore, the present review is intended (i) to provide a comprehensive, up-to-date overview of the ethnomedicinal uses, pharmacological activities, and cultivation of TMM in general and L. rhinocerotis in particular, (ii) to demonstrate how recent scientific findings have validated some of their traditional uses, and (iii) to identify opportunities for future research and areas to prioritize for TMM bioprospecting.
    Matched MeSH terms: Polyporaceae/classification; Polyporaceae/growth & development*; Polyporaceae/chemistry*
  20. Bronchodilator effects of Lignosus rhinocerotis extract on rat isolated airways is linked to the blockage of calcium entry
    Lee MK, Lim KH, Millns P, Mohankumar SK, Ng ST, Tan CS, et al.
    Phytomedicine, 2018 Mar 15;42:172-179.
    PMID: 29655683 DOI: 10.1016/j.phymed.2018.03.025
    BACKGROUND: Lignosus rhinocerotis (Cooke) Ryvarden is a popular medicinal mushroom used for centuries in Southeast Asia to treat asthma and chronic cough. The present study aimed to investigate the effect of this mushroom on airways patency.

    MATERIALS AND METHODS: The composition of L. rhinocerotis TM02 cultivar was analyzed. Organ bath experiment was employed to study the bronchodilator effect of Lignosus rhinocerotis cold water extract (CWE) on rat isolated airways. Trachea and bronchus were removed from male Sprague-Dawley rats, cut into rings of 2 mm, pre-contracted with carbachol before adding CWE into the bath in increasing concentrations. To investigate the influence of incubation time, tissues were exposed to intervals of 5, 15 and 30 min between CWE concentrations after pre-contraction with carbachol in subsequent protocol. Next, tissues were pre-incubated with CWE before the addition of different contractile agents, carbachol and 5-hydroxytrptamine (5-HT). The bronchodilator effect of CWE was compared with salmeterol and ipratropium. In order to uncover the mechanism of action of CWE, the role of beta-adrenoceptor, potassium and calcium channels was investigated.

    RESULTS: Composition analysis of TM02 cultivar revealed the presence of β-glucans and derivatives of adenosine. The extract fully relaxed the trachea at 3.75 mg/ml (p 

    Matched MeSH terms: Polyporaceae/chemistry*
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