Polypore Mushroom Research Articles

Genetic Resources and Mycelial Characteristics of Several Medicinal Polypore Mushrooms (Polyporales, Basidiomycetes)

Mycelial characteristics of dikaryotic collections of 6 medicinal polypore mushrooms (Fomes fomentarius, Fomitopsis pinicola, Ganoderma adspersum, G. applanatum, G. lucidum, and G. resinaceum) with different geographical origins (Armenia, China, France, Iran, Italy, and Russia) were screened. A total of 42 polypore collections were molecularly identified by sequencing the internal transcribed spacer region of the ribosomal RNA genes' cluster, and a phylogenetic tree was constructed. Morphological characteristics of 37 cultures were observed on agar media (malt extract agar, potato dextrose agar) at different temperatures (25, 30, 35, and 38°C) at a pH of 6.0. Colony morphology, pigmentation of mycelium and agar, mycelial growth rate, in vitro teleomorph formation, and other macromorphological characteristics were thoroughly described and illustrated. Micromorphological features of mycelia, such as different hyphal structures, clamp cells, presence and type of asexual sporulation, chlamydospores, and others were observed. The taxonomic significance of the mycelial characteristics revealed was estimated. The obtained results will assist further biotechnological cultivation of medicinal polypore mushrooms to develop novel health care biotechnological products.

Anti-inflammatory effects of Antrodia camphorata, a herbal medicine, in a mouse skin ischemia model

Ethnopharmacological evidenceAntrodia camphorata, a highly valued polypore mushroom native only to Taiwan, has been traditionally used as a medicine for anti-inflammation. Aim of the studyIn this study, anti-inflammatory effects of Antrodia camphorata (AC) and its active compound, ergostatrien-3β-ol (ST1), were investigated in a mouse skin ischemia model induced by skin flap surgery on the dorsal skin. Materials and methodsA U-shaped flap was elevated on the dorsal skin of the nine-week-old male mice. Mice were randomly assigned to six groups for treatment (n=6) including normal skin/propylene glycol (PG), surgical skin flap/PG, solid-state-cultured AC (S/AC), wood-cultured AC (W/AC), high-dose ST1 (H-ST1), low-dose ST1 (L-ST1). Antrodia camphorata was dissolved in 25μL PG and smeared on the skin flap every six hours for 24h. At the end of the experiment, each mouse was anesthetized, and skin tissues were collected from their back for histopathological analysis, extracting RNA and protein according to our previous reports. ResultsSkin-flap-induced ischemia damage significantly increased the expression of the iNOS, COX2, and IL-6 proteins and decreased the expression of IκB protein. In addition, focal, moderate coagulative necrosis with inflammatory cell infiltration was found in the epidermis, and moderate inflammatory cells and necrosis with slight edema was noted in the sub-dermis at 24h after skin flap surgery. However, treatment with solid-state-cultured or wood-cultured AC, or with its derived ST1 active compound, significantly reduced the necrosis and inflammatory cell infiltration in both the epidermis and sub-dermis of the skin flap. The treatments also reduced the inflammatory response by decreasing the expression of inflammation-related genes including iNOS, IL-6, TNF-α, and NF-κB, as shown by changes in RNA and protein expression, when compared with the surgical skin flap procedure alone. ConclusionsThese results demonstrated that methanolic extracts of wood-cultured fruiting bodies and solid-state-cultured mycelia from Antrodia camphorata have excellent anti-inflammatory activities and thus have great potential as an addition for hydrocolloid dressings.

Comparison of solid-state-cultured and wood-cultured Antrodia camphorata in anti-inflammatory effects using NF-κB/luciferase inducible transgenic mice

PurposeAntrodia camphorata (AC), a highly valued polypore mushroom native only to Taiwan, has been traditionally used as a medicine for the treatment of food and drug intoxication, diarrhea, abdominal pain, hypertension, skin itching, and cancer. In this study, both of solid-state-cultured AC (S-AC) and wood-cultured AC (W-AC) were evaluated the anti-inflammatory effects on hyperoxia-induced lung injury in NF-κB-luciferase+/+ transgenic mice. MethodsThe homozygous transgenic mice (NF-κB-luciferase+/+) were randomly assigned to four groups for treatment (n=6) including Normoxia/DMSO group, Hyperoxia/DMSO group, Hyperoxia/S-AC group, and Hyperoxia/W-AC group. After 72h of hyperoxia, we examined the bioluminescence images, reactive oxygen species (ROS), the mRNA and protein expression levels of inflammation factors, and histopathological analyses of the lung tissues. ResultsHyperoxia-induced lung injury significantly increased the generation of ROS, the mRNA levels of IL-6, TNF-α, IL-1β and IL-8, and the protein expression levels of IKKα/β, iNOS and IL-6. Pulmonary edema and alveolar infiltration of neutrophils was also observed in the hyperoxia-induced lung tissue. However, treatment with either S-AC or W-AC obviously decreased hyperoxia-induced generation of ROS and the expression of IL-6, TNF-α, IL-1β, IL-8, IKKα/β and iNOS compared to hyperoxia treatment alone. Lung histopathology also showed that treatment with either S-AC or W-AC significantly reduced neutrophil infiltration and lung edema compared to treatment with hyperoxia treated alone. To find out their major compounds, eburicoic acid and dehydroeburicoic acid were both isolated and identified from S-AC and W-AC by using HPLC, MS, and NMR spectrometry. ConclusionsThese results demonstrated that methanolic extracts both of S-AC and W-AC have excellent anti-inflammatory activities and thus have great potential as a source for natural health products.

Cloning and Molecular Characterization of β-1,3-Glucan Synthase from Sparassis crispa

A β-glucan synthase gene was isolated from the genomic DNA of polypore mushroom Sparassis crispa, which reportedly produces unusually high amount of soluble β-1,3-glucan (β-glucan). Sequencing and subsequent open reading frame analysis of the isolated gene revealed that the gene (5,502 bp) consisted of 10 exons separated by nine introns. The predicted mRNA encoded a β-glucan synthase protein, consisting of 1,576 amino acid residues. Comparison of the predicted protein sequence with multiple fungal β-glucan synthases estimated that the isolated gene contained a complete N-terminus but was lacking approximately 70 amino acid residues in the C-terminus. Fungal β-glucan synthases are integral membrane proteins, containing the two catalytic and two transmembrane domains. The lacking C-terminal part of S. crispa β-glucan synthase was estimated to include catalytically insignificant transmembrane α-helices and loops. Sequence analysis of 101 fungal β-glucan synthases, obtained from public databases, revealed that the β-glucan synthases with various fungal origins were categorized into corresponding fungal groups in the classification system. Interestingly, mushrooms belonging to the class Agaricomycetes were found to contain two distinct types (Type I and II) of β-glucan synthases with the type-specific sequence signatures in the loop regions. S. crispa β-glucan synthase in this study belonged to Type II family, meaning Type I β-glucan synthase is expected to be discovered in S. crispa. The high productivity of soluble β-glucan was not explained but detailed biochemical studies on the catalytic loop domain in the S. crispa β-glucan synthase will provide better explanations.

A comparative study of taxonomy, physicochemical parameters, and chemical constituents of Ganoderma lucidum and G. philippii from Uttarakhand, India

The genus Ganoderma consists of cosmopolitan polypore mushrooms, many of which can cause different types of rots in plants. Many species of this genus are being used for their medicinal and nutraceutical properties in many countries. The present study provides a comparative evaluation of taxonomy, physicochemical parameters, and chemical constituents of Ganoderma lucidum and G. philippii collected from different localities of Uttarakhand, India. The macroscopic and microscopic characters on the basis of which G. lucidum differs from G. philippii include habit, external basidiocarp characteristics, context, pore tube layers and pores, cutis type, and shape and size of basidiospores. The fruiting bodies of both the species were air-dried and ground to powder, which was analyzed for physicochemical parameters and subjected to qualitative chemical screening. The crude powder was subjected to successive Soxhlet extraction for the preparation of various extracts using different solvents. Physicochemical analysis showed variation with respect to foreign matter, moisture content, ash content, extractive values, absorption properties, emulsion properties, foaming properties, dispersibility, and bulk density. Qualitative chemical screening of various extracts showed the presence of carbohydrates, proteins, lipids, glycosides, phenolic compounds, steroids, terpenoids, and saponins in both species.

Трутовые грибы Ботанического сада Петрозаводского государственного университета

Трутовые грибы Ботанического сада Петрозаводского государственного университета

Chlorinated Coumarins from the Polypore Mushroom Fomitopsis officinalis and Their Activity against Mycobacterium tuberculosis

An EtOH extract of the polypore mushroom Fomitopsis officinalis afforded two new naturally occurring chlorinated coumarins, which were identified as the previously synthesized compounds 6-chloro-4-phenyl-2H-chromen-2-one (1) and ethyl 6-chloro-2-oxo-4-phenyl-2H-chromen-3-carboxylate (2). The structures of the two isolates were deduced by ab initio spectroscopic methods and confirmed by chemical synthesis. In addition, an analogue of each was synthesized as 7-chloro-4-phenyl-2H-chromen-2-one (3) and ethyl 7-chloro-2-oxo-4-phenyl-2H-chromen-3-carboxylate (4). All four compounds were characterized physicochemically, and their antimicrobial activity profiles revealed a narrow spectrum of activity with lowest MICs against the Mycobacterium tuberculosis complex.

Bioconversion of Ferulic Acid into High Value Metabolites by White Rot Fungi Isolated from Fruiting-Body of the Polypore Mushroom

The trend toward natural flavors has led to extensive research in this area. In the biotechnological process, ferulic acid was commonly used as a substrate and employs microorganisms as a biocatalyst. In this study, the potential of three white rot fungi- Ganoderma mastoporum, Ganoderma australe and Trametes pavonia - isolated from the fruiting-body of the polypore mushroom and two strains of commercial mushrooms (Ganoderma lucidum and Schizophyllum commune ) were investigated for their ability to convert ferulic acid into its metabolites using GC-MS. The results showed that G. lucidum, S. commune and T. pavonia were able to convert ferulic acid to 4-vinyl guaiacol as a major degradation product, while the extract of G. australe and G. mastoporum grown in the same medium were found to obtain vanillin as a major component. Moreover, various degradation products, including methoxy benzoquinone, 2-methoxyhydroquinone and methyl eugenol, were also found in this experiment. 

Biological and chemical evaluation of anti-TB coumarins from the polypore mushroom, Fomitopsis officinalis

Biological and chemical evaluation of anti-TB coumarins from the polypore mushroom, Fomitopsis officinalis

Generation of a cDNA library reveals the presence of a Ty-1/copia transposon in Phellinus rimosus (Hymenochaetaceae, Basidiomycota)

Lignocellulolytic fungi have significant potential as bio-convertors with a particular capacity to degrade recalcitrant natural composts, such as lignin. We generated a cDNA library of a lignocellulolytic fungus, the Phellinus rimosus (Hymenochaetaceae, Basidiomycota), and a typical species of polypore mushroom from the Brazilian semi-arid region. To the best of our knowledge, this is the first report ever to describe the existence of a Ty1-copia retrotransposon in the P. rimosus species. Given the taxonomic complexity in the Brazilian semi-arid region, our findings will be useful as they provide a molecular identity to this important parasite species of the semi-arid region of Brazil. Key words: Lignocellulolytic fungi, biotechnology, Phellinus rimosus, Ty-1 copia, Brazilian semi-arid region, molecular marker.

The effect of exopolysaccharide biosynthesis and related enzyme activities of Grifola frondosa by the addition of ethanol extracts from traditional Chinese medicine, Gastrodia tuber

Grifola frondosa is an edible medicinal polypore mushroom and its exopolysaccharide (EPS) has demonstrated immunomodulatory effects in many previous studies. After adding the Gastrodia tuber components in culture medium of G. frondosa , both EPS and biomass production were significantly increased. As a result, we detected two enzymes which are related to EPS synthesis. Our results suggested that the activity of α-phosphoglucomutase (α-PGM) increased, while the activity of phosphoglucose isomerase decreased. Mixture components of ethanol extract of Gastrodia tuber was more effective than pure gastrodine. A maximum dry cell weight of 138.5 mg l −1 and the EPS at 0.606 g l -1 were obtained when the unprocessed Gastrodia tuber culture was added. Key words : Exopolysaccharide, phosphoglucomutase (PGM), phosphoglucose isomerase (PGI), Grifola frondosa, Gastrodia tuber .

Protective effect of polysaccharide-protein complex from a polypore mushroom, Phellinus rimosus against radiation-induced oxidative stress

ObjectivesIonizing radiation induces severe oxidative stress in the body resulting an imbalance in prooxidant and antioxidant status in the cell. The aim of the present study is to investigate the protective effect of polysaccharide protein complex (PPC-Pr) isolated from the mushroom Phellinus rimosus against the oxidative stress induced by gamma radiation.MethodologyPPC-Pr complex was isolated from the aqueous extracts of P. rimosus. The complex was administered to Swiss albino mice at a concentration of 5 and 10 mg/kg body weight intraperitoneally for 5 days consecutively and exposed to 4 Gy of gamma irradiation. Animals were sacrificed 1 day after irradiation and the antioxidant parameters such as glutathione, glutathione peroxidase, superoxide dismutase, catalase, glutathione reductase as well as lipid peroxidation were evaluated in both liver and brain tissues to evaluate oxidative stress. Amifostine, a standard radioprotective agent, was used as a positive control. In vitro DNA damage was assessed using the comet assay. Survival studies were also carried out to determine the protective role of PPC-Pr against radiation-induced delayed oxidative stress.ResultsPPC-Pr treatment enhanced the declined levels of antioxidants and comet parameters to a significant level, indicating its antioxidant as well as DNA protecting potential. Significant increase in the survival rate of animals was also observed in irradiated animals treated with PPC-Pr complex. The results were comparable to the standard drug amifostine.DiscussionThe results indicate profound effects of PPC-Pr against radiation-induced oxidative stress. The findings suggest potential therapeutic use of PPC-Pr in radiotherapy.

Ganoderma lucidum and Professor Zhi-Bin Lin

Ganoderma is a genus of polypore mushrooms and includes about 80 species. Ganoderma is characterized by basidiocarps that are large, perennial, woody brackets, also called “conks.” Ganoderma lucidum, also known as lingzhi, is the most commonly cultivated type of Ganoderma in China, and has been used for medical purposes for more than 2,000 years (Figure ​(Figure1).1). Lingzhi is used in China for a great variety of purposes, including asthma, chronic obstructive pulmonary disease, sleep disorder, hepatitis, leucopenia, hypertension, hyperlipidemia, diabetes, and cancer. It is also widely used to promote health in those under a “sub-health” status (Lin, 2011).

My Favorite Enzyme: For love of lectins

I was a student of Fred Smith at the University of Minnesota during 1952–1959. Smith was an Englishman who came to the USA to work on the Manhattan Project to build the atomic bomb. He was an outstanding carbohydrate chemist who was mentored by Sir Norman Haworth, Nobel Laureate in Chemistry for the synthesis of vitamin C (ascorbic acid). Smith was at the University of Birmingham when he received the call to come to the United States to work on ‘‘the bomb’’ because of his expertise in fluorine chemistry, a necessary technique for the separation of the 238 isotope of uranium as the hexafluoride. My work in Smith’s Lab involved studies on periodate oxidation products of glycosides. On receiving my PhD I stayed on as a postdoctoral fellow and instructor for two additional years. Happily for me, I was awarded a Guuggenheim Fellowship which allowed me to spend 2 years in Europe, first with Bill Whelan at the Lister Institute of Preventive Medicine in London, then with Bengt Lindberg in Stockholm. My work in both places involved the isolation, characterization, and chemical and enzymatic synthesis of oligosaccharides from natural sources. These were two wonderful years to work in and experience the scientific and cultural milieu of two different European laboratories. Bill Whelan was an enthusiastic and a helpful mentor. We both smoked at that time, and each morning ‘‘after lighting up,’’ we discussed my research. He always provided insightful and valuable ideas and approaches to pursue. My habit of arising early and getting to the lab in the early morning persisted while in London. At first, this did not go over well with the janitorial staff at the Lister. But after some time we became good friends and they greeted me with much enthusiasm each morning. There was another advantage from arriving early: I had the first use of equipment which was, at times, in short supply. While at the Lister I also had the opportunity of meeting Walter Morgan and his colleague Winifred Watkins. They had been studying the chemistry of human blood group substances for many years and were in a fierce competition with Elvin Kabat, Prof. of Immunochemistry at Columbia University. They were always first to grab a copy of the latest journal to see if Kabat had scooped them. Little did I realize, at the time, that our paths would converge. They did indeed when my lab took up a study of the lima bean lectin which they had used to assay for human type A blood group substance. I had many interesting and fruitful discussions with these colleagues during the following years. My first independent position was at the University of New York at Buffalo. To assist a colleague in the characterization of glycogen from microorganism, I decided to prepare concanavalin A from jack beans (Canavalia ensiformis) which was shown to precipitate glycogen. I followed the procedure of James Sumner who first prepared the protein. I decided to pass the purified protein through Sephadex, crossed-linked dextran, to assess its homogeneity. It did not emerge from the column! In an attempt to displace it from the column I suggested to a graduate student to add glucose. Pure concanavalin A was eluted! Address correspondence to: Irwin J. Goldstein, Department of Biological Chemistry, University of Michigan, Ann Arbor, MI 481095606, USA. E-mail: igoldste@umich.edu Received 12 September 2009; accepted 12 September 2009

Reappearance of the large polypore mushroom, Bondarzewia berkeleyi, in northeastern Kansas

A period of rainy weather in the vicinity of Lawrence, Douglas County, Kansas during the last week of June 2007 facilitated the emergence of a diversity of mushroom species. In early July, a resident of North Lawrence noticed an unusual, white, amorphous mushroom with several finger-like growths on the ground next to a white oak tree (Quercus alba) on his property. On 6 July, the first author visited the site and collected the mushroom which had transformed into an erect, fan-shaped growth with several lobes approximately 12 inches high and 24 inches wide. The mushroom weighed 15 pounds (Fig. 1). The second author identified the specimen as Bondarzewia berkeleyi (Fr.) Bond. et Sing.. Another fruiting body of this species appeared toward the end of June 2008 at the same location on the opposite side of the white oak tree. By the end of July, the mushroom had grown to a total width of 36 inches.

Characterization of neutral and acidic glycosphingolipids from the lectin-producing mushroom, Polyporus squamosus

The polypore mushroom Polyporus squamosus is the source of a lectin that exhibits a general affinity for terminal beta-galactosides, but appears to have an extended carbohydrate-binding site with high affinity and strict specificity for the nonreducing terminal trisaccharide sequence NeuAcalpha2 --> 6Galbeta1 --> 4Glc/GlcNAc. In considering the possibility that the lectin's in vivo function could involve interaction with an endogenous glycoconjugate, it would clearly be helpful to identify candidate ligands among various classes of carbohydrate-containing materials expressed by P. squamosus. Since evidence has been accumulating that glycosphingolipids (GSLs) may serve as key ligands for some endogenous lectins in animal species, possible similar roles for fungal GSLs could be considered. For this study, total lipids were extracted from mature fruiting body of P. squamosus. Multistep fractionation yielded a major monohexosylceramide (CMH) component and three major glycosylinositol phosphorylceramides (GIPCs) from the neutral and acidic lipids, respectively. These were characterized by a variety of techniques as required, including one- and two-dimensional (1)H- and (13)C-nuclear magnetic resonance (NMR) spectroscopy; electrospray ionization-mass spectrometry (ESI-MS, tandem-MS/collision-induced decay-MS, and ion trap-MS(n)); and component and methylation linkage analysis by gas chromatography-mass spectrometry. The CMH was determined to be glucosylceramide having a typical ceramide consisting of 2-hydroxy fatty-N-acylated (4E,8E)-9-methyl-sphinga-4,8-dienine. The GIPCs were identified as Manalpha1 --> 2Ins1-P-1Cer (Ps-1), Galbeta1 --> 6Manalpha1 --> 2Ins1-P-1Cer (Ps-2), and Manalpha1 --> 3Fucalpha1 --> 2Galalpha1 --> 6Galbeta1 --> 6Manalpha1 -->2Ins1-P-1Cer (Ps-5), respectively (where Ins = myo-inositol, P = phosphodiester, and Cer = ceramide consisting mainly of long-chain 2-hydroxy and 2,3-dihydroxy fatty-N-acylated 4-hydroxy-sphinganines). Of these GSLs, Ps-2 could potentially interact with P. squamosus lectin, and further investigations will focus on determining the binding affinity, if any, of the lectin for the GIPCs isolated from this fungus.

Medicinal Polypores of the Forests of North America: Screening for Novel Antiviral Activity

362 International Journal of Medicinal Mushrooms Polypore mushrooms have been used medicinally for thousands of years. Th e Greek physician Dioscorides fi rst described the use of a wood conk, Agarikon, now known as Fomitopsis offi cinalis (Vill.: Fr.) Bond. et Singer (= Laricifomes offi cinalis), as a treatment against consumption in 65 AD. Other wood conks, such as Ling Chi or Reishi, have had a similarly long history of use in Asia. In the past 20 years, wood conks have been carefully explored for their immunomodulating and anticancer properties. More recently, mushrooms, including polypores, have and are being explored for their antimicrobial properties. Upon submitting more than a hundred in vitro cultures of mushrooms to the US Defense Department’s Bioshield BioDefense program, several tests show that some of these polypore mushrooms have strong antiviral activity. Within these verdant natural landscapes, trees hundreds of years old host ancestral strains of these elusive polypores. Species that are now rare, or in some cases thought to be extinct, still reside in the pristine old growth forests of the Pacifi c Northwest of North America. When clones from these mushrooms were grown in vitro and submitted for antiviral screening, several mycelial cultures produced antibiotics eff ective against Pox and other viruses. Notably, strains vary in their antiviral properties. Our natural genomes hold within them great potentials for staving off disease and have not yet been fully explored. Th e fungal diversity within these genomes may prove critical for isolating the most active strains, similar to the lessons learned from the isolation of Penicillium chrysogenum strains that lead to the commercialization of penicillin and saved millions of lives. With deforestation, pollution, and industrialization, societies should reevaluate the importance of their natural forests in the context that they hold within them novel medicines of enormous socioeconomic importance. Th e old paradigm of viewing the forest as valuable only in terms of timber seems overly simplistic given this new knowledge. Medicinal Polypores of the Forests of North America: Screening for Novel Antiviral Activity

Medicinal Polypores of the Forests of North America: Screening for Novel Antiviral Activity

Polypore mushrooms have been used medicinally for thousands of years. The Greek physician Dioscorides first described the use of a wood conk, Agarikon, now known as Fomitopsis officinalis (Vill.: Fr.) Bond. et Singer (= Laricifomes officinalis), as a treatment against consumption in 65 AD. Other wood conks, such as Ling Chi or Reishi, have had a similarly long history of use in Asia. In the past 20 years, wood conks have been carefully explored for their immunomodulating and anticancer properties. More recently, mushrooms, including polypores, have and are being explored for their antimicrobial properties.Upon submitting more than a hundred in vitro cultures of mushrooms to the US Defense Department’s Bioshield BioDefense program, several tests show that some of these polypore mushrooms have strong antiviral activity. Within these verdant natural landscapes, trees hundreds of years old host ancestral strains of these elusive polypores. Species that are now rare, or in some cases thought to be extinct, still reside in the pristine old growth forests of the Pacific Northwest of North America. When clones from these mushrooms were grown in vitro and submitted for antiviral screening, several mycelial cultures produced antibiotics effective against Pox and other viruses. Notably, strains vary in their antiviral properties. Our natural genomes hold within them great potentials for staving off disease and have not yet been fully explored. The fungal diversity within these genomes may prove critical for isolating the most active strains, similar to the lessons learned from the isolation of Penicillium chrysogenum strains that lead to the commercialization of penicillin and saved millions of lives.With deforestation, pollution, and industrialization, societies should reevaluate the importance of their natural forests in the context that they hold within them novel medicines of enormous socio-economic importance. The old paradigm of viewing the forest as valuable only in terms of timber seems overly simplistic given this new knowledge.

Nutritional and Medicinal Potential of Twenty-Three Wild Mushrooms from Northeast Thailand

Twenty-three species of wild mushrooms (mostly polypores from genera Trametes, Ganoderma, Schizophyllum, Inonotus, Phellinus, and Lentinus) from Sakon Nakhon, northeast Thailand, were collected, and their pure cultures were kept at the Department of Plant Pathology, Khon Kaen University, for further studies. In this paper, the nutritional value, potential of exo-polysaccharides production, and ability to produce lectins from all species were investigated. In order to produce mycelial biomass, exo-polysaccharides, and lectins culture was grown in a 250 mL Erlenmeyer flask containing 100 mL of rice bran dextrose malt peptone.(RbDMP) medium at room temperature on a rotary shaker at 125 rpm for 10−15 days. For nutritional value, the mycelial biomass was analyzed for ash, proteins, and Amino, acids contents. The content of ash determined by incineration of mycelia at 500 ± 10°C for 8 hours was very low (approximately 4.73 ± 0.36% of dry matter) in all species. The protein content of all species analyzed through the macro-Kjeldahl method was varied among species, and the average of crude protein was 22.2 ± 5.5% (range 13.3−26.5) of mycelium dry weight. Essential amino acids were determined by using modified techniques (Wiedmeier, Porterfield, and Hendrich, 1982) after hydrolysis of freeze-dried samples with 6 N HC1 for 24 hours at 110°C and analyzed through Shimadzu High Performance Liquid Chromatography. The amino acid patterns were shown clearly, and all amino acids contained in all species were calculated. The amount of the essential amino acids accounted for at least 30.77%, by average, of total amino acids content. Tryptophan could not be detected by the method mentioned above. Essential branch chain amino acids such as isoleucine, leucine, and valine were high, which were useful in treatments involving muscle, mental, and emotional upsets as well as for insommia and nervousness. The amount of exo-polysaccharides was obtained by mixed culture filtrate with 4 vol. of 100% ethanol, stirred vigorously, and kept overnight at 4°C, and the precipitated exo-polysaccharides were air dried and weighted.Exopolysaccharide production varied considerably among species ranging from 0.33 to 6.59 mg/L with an average of 2.91 ± 0.19 mg/L. Lectin extraction was performed in crude extract by using 5 times (w/v), 10 mM phosphate buffer saline (PBS contained 0.02 M sodium bisulphite, pH 7.2), stirred for 2 hours at 4°C, filtrated and centrifuged for 15 minutes at 12,000 rpm. A hemagglutination test of a serial two-fold dilution of the crude extract and cultured broth was investigated by using Microtiter U-plates with 2% suspension of red blood cells of animals (six kinds) in PBS pH 7.2. Hemagglutination titers of lectins were positively detected at 1:1 (majority) up to 1:16 (1 species) in mycelium, cultured broth, or both in most of the species investigated. All measurements clearly indicated the potential of those wild polypore mushrooms from northeast Thailand as good sources of alternative nutritional food and potential uses as alternative medicine to promote good health and to enhance the body's adaptive capabilities.

Purification and Characterization of a Neu5Acα2–6Galβ1–4Glc/GlcNAc-specific Lectin from the Fruiting Body of the Polypore Mushroom Polyporus squamosus

A lectin has been purified from the carpophores of the mushroom Polyporus squamosus by a combination of affinity chromatography on beta-D-galactosyl-Synsorb and ion-exchange chromatography on DEAE-Sephacel. Gel filtration chromatography, SDS-polyacrylamide gel electrophoresis, and N-terminal amino acid sequencing indicated that the native lectin, designated P. squamosus agglutinin, is composed of two identical 28-kDa subunits associated by noncovalent bonds. P. squamosus agglutinin agglutinated human A, B, and O and rabbit red blood cells but precipitated only with human alpha(2)-macroglobulin, of many glycoproteins and polysaccharides tested. The detailed carbohydrate binding properties of the purified lectin were elucidated using three different approaches, i.e. precipitation inhibition assay (in solution binding assay), fluorescence quenching studies, and glycolipid binding by lectin staining on high-performance thin layer chromatography (solid-phase binding assay). Based on the results obtained by these assays, we conclude that although the P. squamosus lectin binds beta-D-galactosides, it has an extended carbohydrate-combining site that exhibits highest specificity and affinity toward nonreducing terminal Neu5Acalpha2, 6Galbeta1,4Glc/GlcNAc (6'-sialylated type II chain) of N-glycans (2000-fold stronger than toward galactose). The strict specificity of the lectin for alpha2,6-linked sialic acid renders this lectin a valuable tool for glycobiological studies in biomedical and cancer research.