In our previous study, active-single compound (ASC), inhibitor of melanogenesis in B16F10 melanoma cell, was identified from Ganoderma lucidum. Thus, the aim of this study is to analyze the change of the ASC contents in G. lucidum when oriental raisin tree (ORT) was added in cultivation of G. lucidum. In HPLC analysis, significantly increased ASC peak was observed in G. lucidum extract supplemented with 15% (v/v) of ORT. In addition, melanogenesis inhibitory effect of G. lucidum extract supplemented with 15% of ORT significantly increased when compared to G. lucidum extract without ORT supplementation. Furthermore, ORT supplementation increased mycelial growth of G. lucidum in both solid and liquid cultivation. These results suggest that oriental raisin tree is useful as natural ingredient for increasing bioactivity including skin-whitening effect and mycelial growth of G. lucidum.

The purpose of this experiment is to measure the change in the growth rate and the amount of bioactive compounds when additives are added to mushrooms. According to the additive content, Ganoderma lucidum ASI7004, Ganoderma lucidum ASI7071, Ganoderma. meredithae KACC42868 and Ganoderma lucidum ASI7013 mycelial growth rate variation analysis, oriental raisin tree, Siberian ginseng, chestnut shell, apple pomace, and Korean cabbage additive increased mycelial growth by approximately 2.5 times when compared to the control group(MCM medium). The results of analyzing the total amount of polyphenol in mycelia of G. lucidum based on different kinds of additives have shown that the amount of polyphenol increased only in G. lucidum ASI7004 with 2% Korean cabbage additive. However, when other additives are added, the amount of polyphenol turned out to be lower than original G. lucidum and other individual additives. The results of analyzing the total amount of triterpenoid in mycelia of G. lucidum based on different kinds of additives have shown that the amount of triterpenoid increased in G. lucidum mycelia with apple pomace and Korean cabbage additives. Siberian ginseng additive was effective in increasing the amount of triterpenoid only when 4% Siberian ginseng was added to G. lucidum ASI7004, yet the increased amount was not significant. Although the addition of additives turned out to be effective to the growth rate, it was not effective enough towards the amount of bioactive compounds. Further experiments are required.

Ganoderma lucidum has a long history for traditional medicine in Asian countries. However, taxonomy of Ganoderma species remains controversial, since it was initially classified on the basis of morphological characteristics. Recently, Ganoderma sichuanense was proposed as a re-proposed name for Ganoderma lucidum in China. Likewise, all of the Korean G. lucidum were clustered into 1 group together with G. sichuanense from China, when a phylogenetic analysis was undertaken based on the ITS rDNA sequences of the Ganoderma species. Furthermore, G. lucidum from Europe and North America were clustered into different group. Based on these results, re-consideration of naming Korean cultivated G. lucidum is needed.

영지(Ganoderma lucidum)는 활엽수의 뿌리 밑동이나 그루터기에서 땅위로 나는데 예전부터 불로초로 알려져온 버섯이다. 각종 무기질과 섬유소, 비타민이 풍부하며 최근들어 항암효과가 인정되고, 고지혈증이나 비만예방 등에 도움을 주는 것으로 알려져 있다. 영지 추출물을 유산균으로 발효하여 당뇨, 활성산소에 의한 노화예방 식품 개발을 목적으로 연구를 수행한 결과는 아래와 같다. 영지 추출물에 유산균(Bifidobacterium bifidum)을 이용하여 1차 발효한 후, 김치 유산균(Lactobacillus sakei subsp.)을 이용하여 2차 발효 하였다. 1차 발효와 2차 발효에 따른 β-glucan, ergothioneine 및 폴리페놀 함량을 측정하고, DPPH free radical 소거활성을 측정하였다. 영지 발효물의 β-glucan함량을 측정한 결과 1차 발효물에서는 1.36 %, 2차발효물에서는 1.62% 로 나타났으며, ergothioneine의 함량은 1차 발효물에서 4.80 mg%, 2차 발효물에서 4.92 mg%로 나타났다. 영지 발효물의 β-glucan과 ergothioneine의 함량은 2차 발효물이 1차 발효물 보다 높게 나타났으며, 폴리페놀 함량을 측정한 결과 1차 발효물에서는 58.53 mg%, 2차 발효물에서는 43.50 mg%로 나타났다. DPPH free radical 소거활성을 측정한 결과 IC50 값이 1차 발효물은 0.41 mg/mL로 나타났으며, 2차 발효물에서는 0.58 mg/mL로 나타났다.

항암, 혈관계질환 개선, 간 보호, 면역력증진, 신경쇠약 완화 등이 있는 것으로 입증되어 있는 영지 균사체를 유산균으로 발효하여 노인건강 음료 및 기능성 식품 소재로 활용하고자 연구를 수행하였다. 영지 균사체 배양액에 유산균(Bifidobacterium bifidum)으로 1차 발효하고, 1차 발효액에 김치 유산균(Lactobacillus sakei subsp.)을 이용하여 2차 발효 하였으며, β-glucan, ergothioneine 및 폴리페놀 함량을 측정하고, DPPH free radical 소거활성을 측정하였다. β-glucan함량을 측정한 결과 영지균사체 배양액 1차 발효물에서는 1.02%, 2차 발효물에서는 0.94%로 나타났으며, ergothioneine의 함량은 1차 발효물에서 2.79 mg%, 2차 발효물에서 2.98 mg%로 나타났다. 폴리페놀 함량을 측정한 결과 1차 발효물에서는 16.35 mg%, 2차 발효물에서는 13.12 mg%로 나타났다. 영지 발효물의 β-glucan 및 폴리페놀의 함량은 1차 발효물이 2차 발효물 보다 높게 나타났으며, ergothioneine 함량은 2차 발효물이 1차 발효물 보다 높게 나타났다. DPPH free radical 소거활성을 측정한 결과 IC50 값이 1차 발효물은 9.91 mg/mL로 나타났으며, 2차 발효물에서는 6.29 mg/mL로 나타났다.

복분자(覆盆子, Rubus coreanus)는 피부노화를 예방하고 고혈압과 동맥경화를 억제해 주는 효과 가 있으며 항암작용과 면역억제작용을 갖는 것으로 보고되었고, 영지(Ganoderma lucidum)는 한방 에서 강장·진해·소종(消腫) 등의 효능이 있어 신경쇠약·심장병·고혈압 등의 치료에 사용한다고 기술 되어있다. 두 소재의 유용성분 활용을 위해 곡물배지(현미, 적미, 흑미)에 복분자 추출물과 부산물을 첨가하여 영지균사의 배양적 특성을 확인하였다. 추출물 농도에 따른 영지균사의 생장율 조사에서는 시험구가 대조구인 곡물배지보다 저조한 생장 을 보였으나 현미에 복분자 추출물 20%를 첨가한 조건에서는 약간 좋은 생장을 보였다. 부산물을 첨가한 조사에서도 시험구가 대조구보다 저조한 생장을 보였으나 현미와 흑미 곡물배지의 경우에는 30%의 부산물을 첨가한 조건에서 균사생장이 가장 양호하였고 적미에서는 첨가농도가 증가함에 따 라 영지균사의 생장이 감소하였다. 본 연구에서는 복분자 추출물을 첨가한 경우 현미(20%), 적미(20%), 흑미(100%) 순으로 균사생장 이 양호하였고, 부산물을 첨가한 경우에는 현미와 흑미는 부산물 30%, 적미는 20%의 부산물을 첨 가한 조건에서 균사생장이 양호하였다. 이중에서 적미에 복분자 부산물 30%를 첨가한 조건에서 영 지균사의 생장이 가장 우수하여 추출물보다는 부산물을 첨가한 조건이 영지균사의 생장이 뛰어난 것을 확인할 수 있었다.

In the present study, the genetic diversity of 69 Ganoderma species from various regions was determined by different molecular markers, including the internal transcribed spacer (ITS) rRNA, a partial β-tubulin gene, and mitochondrial small-subunit ribosomal (SSU) rDNA gene as well as randomly amplified polymorphic DNA (RAPD) analysis. The size of the ITS rDNA regions and mitochondrial SSU rDNA gene from different Ganoderma species varied from 625 to 673 bp and 656 to 2,040bp, respectively, and those of the partial β-tubulin gene sequence were 419 bp. Phylogenetic analysis based on the ITS region, the partial β-tubulin gene, and the mitochondrial SSU rDNA gene reveal that Ganoderma species are largely divided into two groups. Interestingly, most of the Ganoderma lucidum strains could be classified into 1 group, while other Ganoderma species divided into several groups (4 or 5 groups) in phylogenetic tree. One fragment unique to G. lucidum was selected from the RAPD profile and then sequenced. One primer pair (designated as KGS-F and KGS-R) based on this specific fragment was designed to amplify a 559 bp DNA fragment within the sequenced region. A single band with the expected size of 559 bp was observed from G. lucidum, except for G. lucidum strains from China, Canada, and Taiwan. This specific marker for G. lucidum from RAPD analysis, also supported by the phylogenetic analysis of the ITS, partial β-tubulin gene, and mitochondrial SSU rDNA sequences, will be useful for the PCR-based identification of G. lucidum in research applications as well as in the market.

황금은 꿀풀과에 속하는 여러해살이 초본식물인 황금(Scutellaria baicalensis GEORGE)의 뿌 리로 약효성분은 Baicalein, Baicalin, Wogonin 등이 알려져 있다. 영지는 항종양 효과 및 성 인병예방에 효과가 뛰어나 약재로 사용되는 버섯으로 알려져 있다. 최근 친환경 소재에 대한 관심 증대와 더불어 생약초 및 버섯에 대한 관심이 증대하고 있다. 두가지 소재의 유용성분 활용의 극대화를 위해 곡물배지에 황금 추출물과 황금 부산물을 첨가하여 영지버섯 균사체의 배양 특 성을 확인하였다. 모든 시험구에서 황금 추출물 10%를 첨가 하였을 때 가장 좋은 영지 균사 생장을 보였으 며, 최적 부산물 농도는 부산물 20% 첨가한 조건으로 나타났다. 추출물 및 부산물 농도에 비 례하여 영지버섯 균사체의 생장이 억제되는 것을 확인하였다. 흑미에 황금 추출물을 첨가한 조건에서 가장 좋은 영지균사 생장이 관찰되었으며, 부산물은 적미에 황금부산물을 첨가한 조 건에서 가장 좋은 생장을 보였다. 균사체의 생장은 추출물 보다는 부산물을 첨가한 곡물배지 에서 더 좋은 생장을 보였다. 본 연구결과 황금 부산물 20%첨가가 영지균사 배양에 가장 적 합한 조건임을 확인하여, 향후 생약초영지균사 배양에는 황금 20%첨가가 가장 바람직 할 것 으로 보인다. 사 사 : 본 연구는 산업통산자원부 지역특화기술융복합연구지원사업으로 수행되었으며, 이에 감사드립니다.

영지버섯 자실체를 열수추출과 주정추출을 하여 항산화 효능 및 간암 및 위암세포의 생장 저해도를 분석하였다. 항산화 효능을 실시한 결과, 대조군인 Trolox, BHA보다 높은 항산화 효능을 보인 것은 ASI 7004, 7014이며, 이 두 균주는 열수추출물과 주정 추출물에서 모두 높은 항산화 효능을 나타냈다. 나머지 균주는 대조군인 ABTs보다 대체적으로 높은 효능을 보였다. 또한 암세포 생장저해도를 알아보기 위해서 열수, 주정 추출물을 간암세포인 HepG2에 농도별로(100, 200, 400μg/ml) 처리하여, 세포 생장 저해도(MTT assay)를 측정한 결과, 열수 추출물에서는 ASI 7002, 7011, 7014, 7020이 농도 의존적으로 간암세포의 생장을 저해하는 것을 알 수 있었다. 또한 주정 추출물에서는 ASI 7011, 7019가 농도 의존적으로 간암세포 생장을 저해하는 것으로 나타났다. 위암세포인 AGS에 농도별로(100, 200, 400μg/ml)처리하여, 세포 생장 저해도(MTT assay)를 측정한 결과, 열수 추출물에서는 ASI 7001, 7002, 7019, 7020이 농도 의존적으로 위암세포의 생장을 저해하는 것을 알 수 있었다. 또한 주정 추출물에서는 ASI 7001, 7002가 농도 의존적으로 위암세포 생장을 저해하는 것으로 나타났다.

In the present study, a phylogenetic analysis was undertaken based on the internal transcribed spacer (ITS) rDNA and partial β-tubulin gene sequence of the Ganoderma species. The size of the ITS rDNA regions from different Ganoderma species varied from 625 to 673 bp, and those of the partial β-tubulin gene sequence were 419 bp. Based on the results, a phylogenetic tree was prepared which revealed that Korean Ganoderma lucidum strains belong in a single group along with a G. lucidum strain from Bangladesh.

Mushrooms has been used for food ingredients since early times. Recently it is being used as a good pharmaceutical material or functional bio-material. Especially Ganoderma lucidum, Phellinus and Inonotus are used for medicinal purposes. Ganoderma is a basidiomycete white rot fungus which has been used for medicinal purposes for centuries particularly in China, Japan and Korea. A great deal of work has been carried out on Ganoderma lucidum. The metabolites consist of mainly polysaccharides and terpenoids. At present, Ganoderma is a health food supplement to support cancer patients, yet the evidence supporting the potential of direct in vivo anticancer effects should not be underestimated. the aim of this study was to investigate the anti-cancer effects and anti-inflammatory of Ganoderma lucidum in some kinds of human cell lines (Gastric:AGS, liver:HepG2, breast:MCF-7, lung:A549, uterine:Hela, Raw 264.7) we are examined the cell viability by MTT assay and effects of anti-inflammatory by NO assay. Anticancer and anti-inflammatory effect in the 10strains were selected.

Ganoderma lucidum, a popular medicinal mushroom, has been used in China for longevity and health promotion since ancient times. Recently it is being used as a good pharmaceutical material or functional bio-material. Total 106 strains was cultured at temperature(25 ℃, 30 ℃ ,35 ℃) to study cultural properties of Ganoderma lucidum. Then the mycelium growth was digitized in the average diameter. With the result, Ganoderma lucidum strain has the optimum growing temperature at 25 ℃ with 25 strains, 30 ℃ with 70 strains and 35 ℃ with 11 strains. The 84strains were cultivated in wood for investigating the cultivation properties of Ganoderma lucidum. With the result, 70mm~90mm of the pileus diameter (major axis, mm) and 70mm-80mm of pileus diameter (minor axis, mm) was measured after one year cultivation. And 1,000-2,000 ㎟ of strain took the most the area (major axis x minor axis). The 30-40mm of stipe length,1time of harvesting and volume was 200g~400g took the most. 10cm-15cm of the pileus diameter (major axis, mm), 5cm~7.5cm of pileus diameter (minor axis, mm) and 15-20mm of thickness was measured after 2 year cultivation. And 50~100 ㎟ of strain took the most the area (major axis x minor axis). The 5-10mm of stipe length, 2 times of harvesting and volume was 200g~400g took the most. we are selected a middle model through this result. phylogenetic relationships was drawn based on the ITS sequence analysis in Ganoderma lucidum.

Ganoderma lucidum has long been used as a potent medicinal plant. In addition, recent studies showed that G. lucidum is used to prevent or treat various human diseases such as allergy, hepatopathy, hypertension, cancer and diabetes. In this study showed that ethanol extract from G. lucidum have potential inhibitory effects of glycerol 3-phosphat acyltransferase (GPAT) activity and increase of glucose uptake on skeletal muscle cells. Inhibition of GPAT, which catalyze the first step in de novo TAG synthesis, has been proposed as one of the drug targets for insulin resistance and type-2 diabetes. G. lucidum extract inhibited GPAT activity. Bioassay-guided fractionation and isolation of bio-active substances from ethanol extracts of G. lucidum were carried out by using chromatographic techniques and in vitro GPAT enzyme assay. One of bioactive molecules, ergosterol peroxide, was isolated and identified by physical and spectral properties. In this study showed that ethanol extract from G. lucidum and ergosterol peroxide have potential anti- diabetes effects through the increase of glucose uptake. This was as sociated with increased activity AMP-activated protein kinase(AMPK). AMPK is another regulatory protein in the glucose uptake pathway and energy metabolism. The result was that the increase of glucose uptake by G. lucidum extract might be mediated by AMPK.

This study was carried out to investigate characteristic pattern of fruiting body of Ganoderma lucidum and their antioxidant activity. Mycelia of all strains were firstly inoculated into potato dextrose agar(PDA) and then transfered to a media of saw dust which contained 20% rice bran. These mycelia of saw dust were then inoculated into oak tree in polyethylene bags which has been sterilized for 8h at 120℃. The polyethylene bags were sent to a growth room for growth of fruit bodies. Antioxidant activities of each fruiting body were investigated by DPPH method.

Four Ganoderma lucidum strains, Chizhi 05, Jingda, Huizhou and Xinzhou, were screened out as hybrid parent in order to establish G. lucidum cross breeding system that based on protoplast monokaryogenesis method. Monokaryotic strains of each parental strains were obtained and mating type of each monokaryotic strains were determined. One to three monokaryotic strains that have different mating types were mated, and hybrids were identified by clamp connection observation and antagonist response. The results showed that the number of monokaryon came from Chizhi 05, Jingda, Huizhou and Xinzhou was 9, 14, 40 and 38, respectively. Only one mating type was obtained from Jingda, and two mating types were obtained from the other three strains, Chizhi 05, Huizhou and Xinzhou, respectively. Chi-square test showed that the ratio of two mating types of the three strains was 1:1. Fourteen monokaryotic strains of different mating types from 4 parental strains were select as a cross- breeding materia, and 17 hybrids were obtained, which were identified by clamp connection observation and antagonist response. This study proclaimed that the practicality of the hybridization breeding of G. lucidum by protoplast monokaryogenesis method.