• Mycobiology
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• v.30 no.2
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• pp.82-87
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• 2002

Species of Phellinus were known to harmful fungi causing white pocket rot and severe plant disease such as canker or heartrot in living trees in the West, but some species have been used to traditional medicines in the Orient for a long time. In this study the partial D1-D2 nucleotide sequences of 28S ribosomal DNA from 13 Phellinus strains were determined and compared with the sequences of 21 strains obtained from GenBank database. According to the neighbor-joining(NJ) method comparing the sequence data the phylogenetic tree was constructed. The phylogenetic tree displayed the presence of four groups. Group I includes P. ferreus, P. gilvus and P. johnsonianus, Group II contains P. laevigatus, P. conchatus and P. tremulae, Group III possesses P. linteus, P. weirianus, P. baumii, P. rhabarbarinus and P. igniarius, and Group IV comprises P. pini, P. chrysoloma. P. linteus and P. baumii, which were used mainly in traditional medicine, belong to the same group, but exactly speaking both were split into two different subgroups. To detect P. linteus only, we developed the PCR primer, D12HR. The primer showed the specific amplification of P linteus, which is permitted to medicinal mushroom in the East. The results make a potential to be incorporated in a PCR identification system that could be used for the rapid identification of this species from its related species, P. linteus especially.

• Journal of Life Science
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• v.23 no.11
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• pp.1388-1396
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• 2013

This study used an S-180 cell-injected mouse model to evaluate the antitumor effects of the acute and subacute toxicity of Keumsa (Phellinus linteus) extract intravenously administrated in ICR mice. When administered intravenously (31.3-250 mg/kg body weight), Keumsa (Phellinus linteus) extract significantly inhibited the growth of the solid tumor cell. The antitumor activity of Keumsa (Phellinus linteus) extract increased in a dose-dependent manner. The highest dose (250 mg/kg body weight) was highly effective, reducing tumor formation by 42.7% compared with the control group. In the acute toxicity test, $LD_{50}$ of the Keumsa (Phellinus linteus) extract showed 632.84 mg/kg (♂) and 814.48 mg/kg (♀) after intravenous administration. In addition, liver and spleen weight were increased in a dose-dependent manner. In the subacute toxicity test, the mice were intravenously administered over the course of 28 days. The $LD_{50}$ of the Keumsa (Phellinus linteus) extract showed 355.41 mg/kg (♂) and 383.53 mg/kg (♀) after intravenous administration. The liver and spleen weight also increased in a dose-dependent manner. In the case of the group that received more than 125 mg/kg of intravenous administration, exercise capacity, such as jumping ability and agility, were significantly increased. These results suggest that Keumsa (Phellinus linteus) extract can be regarded as a potent enhancer of the innate immune response, and it can be considered as a new natural product with low toxicity that may be used as a candidate for antitumor action.

• The Korean Journal of Mycology
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• v.41 no.4
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• pp.243-247
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• 2013

Novel Phellinus linteus HN1009K (HK1009) has been characterized on speedy mycelial growth and yearly artificial cultivation. In this study, antioxidant and anti-inflammatory activities of ethanol extract from mycelium and fruiting body of P. linteus HN1009K (HK1009) were compared to those of P. linteus (PL, Korea Sangwhang) and P. baumii (PB, Jangsu Sanghwang) as controls. HK1009 extract showed higher DPPH free radical scavenging activity in both mycelial and fruiting body samples than those of PL and PB. However, in xanthine oxidase assay, antioxidant effect was highly measured on mycelial samples of PL and HK1009. Mycelial extract of HK1009 induced high inhibitory effect on LPS-induced nitric oxide (NO) production in RAW264.7 cells comparing to PL and PB without toxicity on the cells. The results suggest that P. linteus HN1009K possesses superior antioxidant activity and anti-inflammatory effects.

• Mycobiology
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• v.36 no.3
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• pp.178-182
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• 2008

In order to increase the mycelial production of Phellinus linteus, which exhibits potent anticancer activity, some ingredients of the medium used to culture P. linteus were investigated. The optimal medium composition for the production of Phellinus linteus was determined to be as follows: fructose, 40 g/l; yeast extract, 20 g/l; $K_2HPO_4$, 0.46 g/l; $K_2HPO_4$, 1.00 g/l; M$MgSO_4\cdot7H_2SO$, 0.50 g/l; $FeCl_2\cdot6_2O$, 0.01 g/l; $MnCl_2\cdot4H_2O$, 0.036 g/l; $ZnCl_2$, 0.03 g/l; and $SuSO_4\cdot7H_2O$, 0.005 g/l. The optimal culture conditions were determined to be as follows: temperature, 28$^{\circ}C$; initial pH, 5.5; aeration, 0.6 vvm; and agitation, 100 rpm, respectively. Under optimal composition and conditions, the maximum mycelial biomass achieved in a 5 l jar fermentor was 29.9 g/l.

• Mycobiology
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• v.30 no.4
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• pp.197-201
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• 2002

Specific primer sets based on ribosomal DNA(rDNA) internal transcribed specer(ITS) sequences were designed for rapid detection of Phellinus linteus and P. baumii. Polymerase chain reaction(PCR) with these primers produced unique bands for each Phellinus species. The annealing temperature range is from $40^{\circ}C\;to\;55^{\circ}C$. The length of PCR products(P. linteus and P. baumii) using designed combinative primer sets of PL1F, PL2R, PB1F, PB2R, ITS5F and ITS4R, were from 520 by to 730 bp. Fifteen strains of Phellinus species including P. linteus, P. baumii, P. weirianus, P. johnsonianus, P. rhabarberinus, P. pini, P. gilvus, P. igniarius, P. nigricans and P. laevigatus were examined in this study. Five strains, including two isolated strains of P. linteus(MPNU 7001 and MPNU 7002), and two isolated strains of P. baumii(MPNU 7004 and MPNU 7005) were shown to have about 520 bp (PL1F-PL2R), 700 bp (TTS5F-PL2R) and 600 bp (PB1F-ITS4R) -sized PCR single bands respectively. This molecular genetic technique provided a useful method for rapid detection and identification of P. linteus and P. baumii.

• The Korean Journal of Mycology
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• v.38 no.1
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• pp.57-61
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• 2010

For development of anti-obesity nutraceuticals from mushrooms, new anti-obesity lipase inhibitor from Phellinus linteus was purified by systematic solvent extraction, TLC and HPLC and characterized. Methanol extract from P. linteus most effectively inhibited(72.5%) porcine pancreatic lipase and ethyl acetate fraction showed the highest inhibitory activity in the systematic solvent extraction. A lipase inhibitor from the ethyl acetate fraction was purified through following steps including 3 times silica gel chromatography and preparative HPLC. The purified lipase inhibitor was a yellowish geen powder and its $IC_{50}$ value was 175 ng. Its molecular weight by MALDI-TOF-MS was 523.06 Da and showed maximal absorption spectrum at 225.1 nm.

• Mycobiology
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• v.31 no.3
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• pp.133-138
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• 2003

Analysis of phylogenetic relationship was performed among Phellinus species based on 18S ribosomal subunit sequence data. Twenty-five strains of 19 Phellinus species including P. linteus were examined in this study. Regions of 18S ribosomal subunit were very conserved, but some variable regions between Phellinus species were observed. The species-specific detection primers, modified by 2 or 3 nucleotides in sense primer were designed based on 18S ribosomal DNA(rDNA) sequence data. The 210 by PCR bands were detected with annealing temperature $48^{\circ}C$. The 18S 2F-18S 4R detection primer set distinguished P. linteus from various Phellinus species but some species like P. baumii, P. weirianius, P. rhabarberinus and P. pomaceus also had weak reactivity on this primer set. The 18S 3F-18S 4R primer set distinguished only P. linteus from various Phellinus species, although sensitivity with this primer set was lower than that of 18S 2F-18 4R primer set. These primer sets would be useful for the detection of only P. linteus among unknown Phellinus species rapidly.

• Mycobiology
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• v.38 no.1
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• pp.52-57
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• 2010

To develop a potent anti-obesity lipase inhibitor from mushroom, the lipase inhibitory activities of various mushroom extracts were determined. Methanol extracts from Phellinus linteus fruiting body exhibited the highest lipase inhibitory activity (72.8%). The inhibitor was maximally extracted by treatment of a P. linteus fruiting body with 80% methanol at $40^{\circ}C$ for 24 hr. After partial purification by systematic solvent extraction, the inhibitor was stable in the range of $40\sim80^{\circ}C$ and pH 2.0~9.0. In addition to lipase inhibitory activity, the inhibitor showed 59.4% of superoxide dismutase-like activity and 56.3% of acetylcholinesterase inhibitory activity.

• Korean Chemical Engineering Research
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• v.47 no.2
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• pp.220-229
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• 2009

Strain improvement and morphology investigation in bioreactor cultures were undertaken in suspended cultures of Phellinus linteus mycelia for mass production of protein-bound polysaccharides(soluble ${\beta}$-D-glucan), a powerful immuno-stimulating agent. Phellineus sp. screened for this research was identified as Phellinus linteues through ITS rDNA sequencing method and blast search, demonstrating 99.7% similarity to other Phellinus linteus strains. Intensive strain improvement program was carried out by obtaining large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because large numbers of protoplasts ($1{\times}10^5{\sim}10^6\;protoplasts/ml$) formed using the banding filtration method with the cell wall-disrupting enzymes could be regenerated in relatively high regeneration frequency($10^{-2}{\sim}10^{-3}$) in the newly developed regeneration medium. It was demonstrated that the strains showing high performances in the protoplast regeneration and solid growth medium were able to produce 5.8~6.4%(w/w) of ${\beta}$-D-glucan and 13~15 g/L of biomass in stable manners in suspended shake-flask cultures of P. linteus mycelia. In addition, cell mass increase was observed to be the most important in order to enhance ${\beta}$-D-glucan productivity during the course of strain improvement program, since the amount of ${\beta}$-D-glucan extracted from the cell wall of P. linteus mycelia was almost constant on the unit biomass basis. Therefore we fully investigated the fungal cell morphology, generally known as one of the key factors affecting cell growth extent in the bioreactor cultures of mycelial fungal cells. It was found that, in order to obtain as high cell mass as possible in the final production bioreactor cultures, the producing cells should be proliferated in condensed filamentous forms in the growth cultures, and optimum amounts of these filamentous cells should be transferred as active inoculums to the production bioreactor. In this case, ideal morphologies consisting of compacted pellets less than 0.5mm in diameter were successfully induced in the production cultures, resulting in shorter period of lag phase, 1.5 fold higher specific cell growth rate and 3.3 fold increase in the final biomass production as compared to the parallel bioreactor cultures of different morphological forms. It was concluded that not only the high-yielding but also the good morphological characteristics led to the significantly higher biomass production and ${\beta}$-D-glucan productivity in the final production cultures.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.1
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• pp.131-135
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• 2007

Various Phellinus species (Hymenochaetaceae) including P. linteus, P. baumii, P. pini, and P. igniarius of domestic and foreign products, were examined by gas chromatography to validate their composition of crude polysaccharides. The crude polysaccharide fraction, which is known to be one of the physiologically active fractions, were prepared by a cold ethanol precipitation method and the yield of the crude polysaccharide fractions of P. linteus, P. baumii, P. pini, and P. igniarius were 15.0%, 5.1%, 6.3% and 5.7%, respectively. The total monosaccharide amount of the crude polysaccharide fraction of P. linteus was 148.713 mg/g (dry weight of mushroom) while those of P. baumii, P. pini and P. igniarius were 29.022 mg/g, 29.222 mg/g and 1.546 mg/g, respectively. Although the major monosaccharide of all Phellinus mushrooms was glucose, there were remarkable differences in the absolute quantity of each monosaccharide. P. linteus included plenty of monosaccharides except mannose and ribose in comparison with P. baumii and P. pini. The absolute quantities of monosaccharides of P. baumii and P. pini were nearly similar except arabinose. The total monosaccharides of P. igniarius were minimal, but the portions of galactose and mannose in P. igniarius were greater than in the other species.