Kuala Selangor, Malaysia
Kuala Selangor, Malaysia

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Yap H.-Y.Y.,University of Malaya | Fung S.-Y.,University of Malaya | Tan N.-H.,University of Malaya | Ng S.-T.,Ligno Biotech Sdn. Bhd. | Tan C.-S.,The Malaysian Agricultural Research and Development Institute MARDI
International Journal of Agriculture and Biology | Year: 2014

Two new species of the tiger milk mushroom, namely Lignosus tigris and L. cameronensis, were reported recently based on studies on morphological characteristics. Both Lignosus species are known to be medicinal fungi and have potential to be used as functional food. Since intact fungal samples of the two Lignosus species are not always available for morphology-based identification, a DNA barcode marker approach based on the internal transcribed spacer (ITS) region via PCR technology has been developed. The DNA markers are highly specific and easily amplified, thus allowing rapid identification and reliable authentication of these two Lignosus species. Identification and authentication of the fungi are important in view of the potential medicinal and functional food applications of the two fungi. © 2014 Friends Science Publishers.


Yap H.-Y.Y.,University of Malaya | Chooi Y.-H.,Australian National University | Firdaus-Raih M.,National University of Malaysia | Fung S.-Y.,University of Malaya | And 3 more authors.
BMC Genomics | Year: 2014

Background: The sclerotium of Lignosus rhinocerotis (Cooke) Ryvarden or Tiger milk mushroom (Polyporales, Basidiomycota) is a valuable folk medicine for indigenous peoples in Southeast Asia. Despite the increasing interest in this ethnobotanical mushroom, very little is known about the molecular and genetic basis of its medicinal and nutraceutical properties.Results: The de novo assembled 34.3 Mb L. rhinocerotis genome encodes 10,742 putative genes with 84.30% of them having detectable sequence similarities to others available in public databases. Phylogenetic analysis revealed a close evolutionary relationship of L. rhinocerotis to Ganoderma lucidum, Dichomitus squalens, and Trametes versicolor in the core polyporoid clade. The L. rhinocerotis genome encodes a repertoire of enzymes engaged in carbohydrate and glycoconjugate metabolism, along with cytochrome P450s, putative bioactive proteins (lectins and fungal immunomodulatory proteins) and laccases. Other genes annotated include those encoding key enzymes for secondary metabolite biosynthesis, including those from polyketide, nonribosomal peptide, and triterpenoid pathways. Among them, the L. rhinocerotis genome is particularly enriched with sesquiterpenoid biosynthesis genes.Conclusions: The genome content of L. rhinocerotis provides insights into the genetic basis of its reported medicinal properties as well as serving as a platform to further characterize putative bioactive proteins and secondary metabolite pathway enzymes and as a reference for comparative genomics of polyporoid fungi. © 2014 Yap et al.; licensee BioMed Central Ltd.


Lee S.S.,University of Malaya | Tan N.H.,University of Malaya | Fung S.Y.,University of Malaya | Sim S.M.,University of Malaya | And 2 more authors.
BMC Complementary and Alternative Medicine | Year: 2014

Background: The sclerotium of Lignosus rhinocerotis (Cooke) Ryvarden (Tiger Milk mushroom) is used as a traditional medicine to relieve cough, asthma and chronic hepatitis. The traditional uses of the sclerotium are presumably related to its anti-inflammatory effect. The present study was carried out to evaluate the anti-inflammatory activity of the sclerotial powder of L. rhinocerotis (Cooke) Ryvarden (Tiger Milk mushroom) cultivar TM02.Methods: The anti-acute inflammatory activity of the sclerotial powder of L. rhinocerotis cultivar TM02 was investigated using carrageenan-induced paw edema test while the inhibition of transudative and proliferative phases of chronic inflammation were studied by cotton pellet induced granuloma model. Sprague Dawley rats were used in both studies. The anti-inflammatory activity was also measured by inhibition of lipopolysaccharide induced TNF-alpha production in RAW 264.7 macrophage cells.Results: Cold water extract (CWE), hot water extract (HWE) and methanol extract (ME) of the sclerotial powder of L. rhinocerotis cultivar TM02 possessed anti-acute inflammatory activity as was measured by carrageenan-induced paw edema test, with CWE being the most potent. The acute anti-inflammatory activity of the cold water extract (CWE) was mainly contributed by its high molecular weight (HMW) fraction isolated by Sephadex G50 gel filtration chromatography. Its protein component was very potent in the inhibition of TNF-alpha production with an IC50 of 0.76 μg/ml. CWE at 200 mg/kg did not inhibit transudative and proliferative phase of chronic inflammation as shown by using the cotton pellet induced granuloma model.Conclusions: Our results suggested that most of the bioactive substance(s) contributed to the acute-inflammatory activity of the sclerotial powder of L. rhinocerotis cultivar TM02 appear to be in the CWE, particularly its HMW fraction. The anti-inflammatory activity of CWE was mainly contributed by the protein component of the high molecular weight (HMW) fraction and it exhibited strong inhibitory effect on TNF-alpha production but the possibility of synergistic effect between HMW, MMW and LMW fractions cannot be excluded. Future studies will provide new insights into the anti-inflammatory activity of the sclerotial powder. © 2014 Lee et al.; licensee BioMed Central Ltd.


Yap H.-Y.Y.,University of Malaya | Aziz A.A.,University of Malaya | Fung S.-Y.,University of Malaya | Ng S.-T.,Ligno Biotech Sdn. Bhd | And 2 more authors.
International Journal of Medical Sciences | Year: 2014

The Lignosus is a genus of fungi that have useful medicinal properties. In Southeast Asia, three species of Lignosus (locally known collectively as Tiger milk mushrooms) have been reported including L. tigris, L. rhinocerotis, and L. cameronensis. All three have been used as important medicinal mushrooms by the natives of Peninsular Malaysia. In this work, the nutritional composition and antioxidant activities of the wild type and a cultivated strain of L. tigris sclerotial extracts were investigated. The sclerotia are rich in carbohydrates with moderate amount of protein and low fat content. Free radical scavenging activities of L. tigris sclerotial extracts correlate with their phenolic content, which ranges from 6.25 to 45.42 mg GAE/g extract. The FRAP values ranged from 0.002 to 0.041 mmol/min/g extract, while the DPPH•, ABTS•+, and superoxide anion (SOA) scavenging activities ranged from 0.18 to 2.53, 0.01 to 0.36, and -4.53 to 10.05 mmol Trolox equivalents/g extract, respectively. L. tigris cultivar shows good prospect to be developed into functional food due to its good nutritional value and potent SOA scavenging activity. © Ivyspring International Publisher.


Lee M.L.,University of Malaya | Tan N.H.,University of Malaya | Fung S.Y.,University of Malaya | Tan C.S.,Biotechnology Research Center | Ng S.T.,Ligno Biotech Sdn Bhd
Evidence-based Complementary and Alternative Medicine | Year: 2012

Lignosus rhinocerus, the tiger milk mushroom, is one of the most important medicinal mushrooms used by the indigenous people of Southeast Asia and China. It has been used to treat breast cancer. A cold water extract (LR-CW) prepared from the sclerotia of L. rhinocerus cultivar was found to exhibit antiproliferative activity against human breast carcinoma (MCF-7) and human lung carcinoma (A549), with IC 50 of 96.7μg/mL and 466.7μg/mL, respectively. In comparison, LR-CW did not show significant cytotoxicity against the two corresponding human normal cells, 184B5 (human breast cell) and NL 20 (human lung cell). DNA fragmentation studies suggested that the cytotoxic action of LR-CW against cancer cells is mediated by apoptosis. Sephadex G-50 gel filtration fractionation of LR-CW yielded a high-molecular-weight and a low-molecular-weight fraction. The high-molecular-weight fraction contains mainly carbohydrate (68.7) and small amount of protein (3.6), whereas the low-molecular-weight fraction contains 31 carbohydrate and was devoid of protein. Only the high-molecular-weight fraction exhibited antiproliferative activity against cancer cells, with IC 50 of 70.0μg/mL and 76.7μg/mL, respectively. Thus, the cytotoxic action of the LR-CW is due to the high-molecular-weight fraction, either the proteins or protein-carbohydrate complex. Copyright 2012 M. L. Lee et al.


Yap Y.H.,University of Malaya | Tan N.,University of Malaya | Fung S.,University of Malaya | Aziz A.A.,University of Malaya | And 2 more authors.
Journal of the Science of Food and Agriculture | Year: 2013

Background: Lignosus rhinocerus (tiger milk mushroom) is an important medicinal mushroom used in Southeast Asia and China, and its sclerotium can be developed into functional food/nutraceuticals. The nutrient composition, antioxidant properties, and anti-proliferative activity of wild type and a cultivated strain of L. rhinocerus sclerotia were investigated. Results: The sclerotial powder has high carbohydrate but low fat content. Interestingly, the cultivated strain contains higher amounts of protein and water-soluble substances than the wild type. Phenolic content of hot-water, cold-water, and methanol extracts of the sclerotial powders ranged from 19.32 to 29.42mg gallic acid equivalents g-1 extract, while the ferric reducing antioxidant power values ranged from 0.006 to 0.016mmolmin-1g-1 extract. The DPPH•, ABTS•+, and superoxide anion radical scavenging activities of the extracts ranged from 0.52 to 1.12, 0.05 to 0.20, and -0.98 to 11.23mmol Trolox equivalents g-1 extract, respectively. Both strains exhibited strong superoxide anion radical scavenging activity comparable to rutin. The cold-water extracts exhibited anti-proliferative activity against human breast carcinoma (MCF-7) cells, with IC50 values of 206μgmL-1 and 90μgmL-1 for the wild type and cultivated strains, respectively. Conclusion: The cultivated L. rhinocerus sclerotium has the potential to be developed into functional food/nutraceuticals. © 2013 Society of Chemical Industry.


Tan C.-S.,Malaysian Agricultural Research and Development Institute MARDI | Ng S.-T.,LiGNO Biotech Sdn Bhd | Tan J.,University of Malaya
Mycotaxon | Year: 2013

Lignosus cameronensis and L. tigris are described as new based on collections from the tropical forests of Pahang, Malaysia. Phenotypic and genotypic data support both as new species. Morphological features are illustrated, and the associated Internal Transcribed Region (ITS1 + 5.8S + ITS2) rDNA sequences have been deposited in the GenBank. Pore and basidiospore sizes are the main characters distinguishing these two Lignosus species from other members of the genus, with L. tigris distinguished from L. sacer by its larger pore size and smaller basidiospores and L. cameronensis separated from L. ekombitii by its smaller basidiospores. A key to the species of Lignosus is provided. © 2013. Mycotaxon, Ltd.


Yap H.-Y.Y.,University of Malaya | Fung S.-Y.,University of Malaya | Ng S.-T.,Ligno Biotech Sdn. Bhd | Tan C.-S.,Ligno Biotech Sdn. Bhd | Tan N.-H.,University of Malaya
International Journal of Medical Sciences | Year: 2015

Lignosus rhinocerotis (Cooke) Ryvarden (Polyporales, Basidiomycota), also known as the tiger milk mushroom, has received much interest in recent years owing to its wide-range ethnobotanical uses and the recent success in its domestication. The sclerotium is the part with medicinal value. Using two-dimensional gel electrophoresis coupled with mass spectrometry analysis, a total of 16 non-redundant, major proteins were identified with high confidence level in L. rhinocerotis sclerotium based on its genome as custom mapping database. Some of these proteins, such as the putative lectins, immunomodulatory proteins, superoxide dismutase, and aegerolysin may have pharmaceutical potential; while others are involved in nutrient mobilization and the protective antioxidant mechanism in the sclerotium. The findings from this study provide a molecular basis for future research on potential pharmacologically active proteins of L. rhinocerotis. © Ivyspring International Publisher.


PubMed | University of Malaya and Ligno Biotech Sdn Bhd
Type: | Journal: Journal of ethnopharmacology | Year: 2015

The sclerotium of Lignosus rhinocerotis (Cooke) Ryvarden (tiger milk mushroom) has been traditionally used as a complementary and alternative medicine for cancer treatment by the local communities of Southeast Asia. Despite the continuous research interest in its antiproliferative activity, the identity of the bioactive compound(s) responsible has yet to be determined. This study aims to bridge the gap in existing research literature by using proteomics approach for investigation of the nature of the anticancer substance of L. rhinocerotis.To elucidate the proteome of L. rhinocerotis TM02 sclerotium by protein mass spectrometry and to further isolate and identify the cytotoxic component(s) bearing anticancer potential.The proteome of L. rhinocerotis sclerotium was analyzed by label-free quantitative shotgun proteomics, using 1D-SDS-PAGE coupled with nano-ESI-LC-MS/MS based on the availability of its genome-sequence database. The cytotoxicity of L. rhinocerotis sclerotial extracts against human breast adenocarcinoma cells (MCF7) were assessed by MTT cytotoxicity assay prior to successive purification steps by a combination of gel filtration chromatography, ammonium sulfate precipitation, and anion exchange chromatography. Bioactive compound(s) in the extracts was identified by shotgun proteomics and N-terminal protein sequencing.Several proteins with interesting biological activities including lectins, fungal immunomodulatory proteins, and several antioxidant proteins were identified from the proteome of L. rhinocerotis. A cytotoxic protein fraction (termed F5) which was partially purified from its sclerotial cold water extract F5 shows two distinct bands of 31 and 36 kDa in reducing SDS-PAGE and exhibited potent selective cytotoxicity against MCF7 cells with IC50 value of 3.00 1.01 g/ml. Both bands were identified to be serine protease by LC-MS/MS analysis. Phenylmethylsulfonyl fluoride, a specific serine protease inhibitor, inhibited both the proteolytic activity and cytotoxicity of F5, suggesting that the cytotoxicity of F5 is related to its protease activity.This study provides the first comprehensive and semi-quantitative profiling of the proteome of L. rhinocerotis sclerotium. Further investigation into its selective cytotoxicity shows that a serine protease-like protein, termed F5, may be targeted for new anticancer agent development.


PubMed | University of Malaya and Ligno Biotech Sdn. Bhd.
Type: Comparative Study | Journal: Nutrition research (New York, N.Y.) | Year: 2016

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.

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