Recently, active research in Korea and worldwide has begun to focus on gene function and cultivar development using gene editing tools. This research, in addition to studies on edible mushroom, aims to enhance the physical and biochemical characteristics of mushrooms for applications in materials and substance production. For these studies, efficient isolation of protoplasts from the target mushroom is critical. However, several commercial cell wall-lysing enzyme cocktails, including Novozyme234, Glucanex, and Lysing enzymes, have recently been discontinued. In this study, we aimed to identify alternative enzyme systems to replace the discontinued cell wall-lysing enzymes for stable isolation of protoplasts from Ganoderma lucidum. To select an optimal osmotic buffer, enzyme function in 0.6 and 1.2 M Sorbitol, Sucrose, Mannitol, and KCl was assessed. The effect of reaction time was also evaluated. Protoplast isolation efficiency of each alternative enzyme was tested using lysing enzymes from Trichoderma harzianum, Chimax-N, and Yatalase, either individually or in combination. This matrix of studies identified enzymes and optimal conditions that could replace the discontinued lysing enzymes.

Recently, with the increasing global demand for Ganoderma lucidum as a material for functional beverages, varieties with enhanced functionality are needed. As part of this effort, 10 strains were selected from a collection of 160 G. lucidum strains, based on their quantity and fruiting body characteristics. These included four strains collected in Korea, three in Japan, and three of unknown origin. Approximately 44% of the 1,290 hybrid strains were secured. After two rounds of bag cultivation in oak sawdust, three strains (G-20-4, G-20-5, and G-20-7) were ultimately selected for single-timbered cultivation. Among these, G-20-4 demonstrated the earliest harvesting with a large size and a bright cap. Additionally, this strain exhibited the highest levels of total glucan, β-glucan, total triterpenoid, total polyphenol, and total flavonoid content.

High-income mushroom crops require complex farming. Therefore, we conducted a test to identify the optimum temperature for the production of antler-shaped Ganoderma lucidum using the King Oyster mushroom cultivation facility. T-N showed 0.28% of oak sawdust and 2.2% of nutritional source. The pH of oak sawdust was 6.0, indicating weak acidity, and that of rice steel was 6.6, indicating neutrality in nutrition source. Study on the quality characteristics of mushrooms showed that the number of days at 25°C were 5~6 and those at 30°C were 3~5; the representative length at 25°C was 57.5 mm and that at 30°C was 92.2 mm; the biological weight at 25 °C was 43 g, which was greater than that at 30°C.

염증은 외부 자극으로부터 보호하는 중요한 면역반응이다(Jeong et al., 2012). 그러나 NO 및 inflammatory cytokine과 같은 염증 매개 인자의 과도한 생성은 비정상적인 염증 반응을 초래할 수 있다(Paradise et al., 2010). 염증 매개 인자의 생성과 염증 신호 전달을 조절하는 중요한 역할을 하는 대식세포는 LPS 자극에 의해 NF-κB가 활성화 되어 inflammatory cytokine 등의 염증 매개 인자들이 분비가 증가되며 염증 반응을 심화시킨다(Lee et al., 2012). 본 연구에서는 영지버섯균 발효 꾸지뽕나무 가지 톱밥 추출물을 이용하여 LPS로 염증을 유도한 Raw264.7세포에서 염증 관련 인자들의 생성 및 발현에 미치는 영향을 분석하여 항염증 효과를 확인하였다. NO는 염증에서 중요한 역할을 하며 대식세포의 염증 반응 조절 평가시 대표적인 지표로 사용되며 iNOS에 의해 생성이 유도되며 PGE2는 염증 매개 인자로 inflammatory cytokine 생성에 관여하며 COX-2에 의해 생성이 유도된다 (Paradise et al., 2010; Posadas et al., 2000). 발효추출물 이 RAW264.7세포에서 세포독성 없이 NO의 생성과 PGE2의 생성을 감소시켰으며 이 결과는 발효 추출물 처리에 의해 iNOS와 COX-2의 발현이 감소한 것과 일치하 였다. 또한 염증 반응을 조절하는 대표적인 inflammatory cytokine으로 알려진 IL-1β, TNF-α의 생성이 감소되었다. 따라서 발효추출물은 NO, PGE2, inflammatory cytokines 생성을 감소시켜 항염증 효과를 나타낸다고 볼 수 있다. NF-κB는 iNOS, COX-2 및 inflammatory cytokine의 발현을 조절하는 전사인자로 IκB와의 결합에 의해 NF-κB의 핵 내 이동이 억제되며 불활성화상태로 존재한다. LPS 자극에 의해 IκB가 인산화 및 분해되면 NF-κB가 활성화 되어 핵 내로 이동하여 염증성 매개인자들의 발현을 유도 하고, 염증성 질환 뿐만 아니라 다양한 질환을 유발시키는 것으로 알려져 있다 (Kawai and Akira, 2006; Ghosh and Ksarin, 2002). 본 연구에서는 LPS에 의해 자극된 RAW264.7 세포에서 IκB의 분해 및 NF-κB의 전이가 발 효추출물에 의해 감소됨을 확인하였으며 발효추출물에 의한 iNOS, COX-2 및 inflammatory cytokine 감소는 NF- κB 활성화 감소에 의해 조절되는 것으로 사료된다. 이러한 결과들을 통해서 발효추출물이 NF-kB 활성화 억제를 통해 염증 매개 인자들의 생성 및 발현을 감소시킴으로서 염증 반응 억제 효과를 나타냄을 확인하여, 염증 관련 질환에 대해 예방 및 개선을 위한 항염증 기능성 소재로 사용될 수 있음을 시사한다.

The purpose of this study was to observe the effects of the polysaccharide (GLP) obtained from the liquid cultured Ganoderma lucidum on the lipidperoxidation in a rat liver microsome and hepatotoxicity in the primary cultured rat hepatocytes. It is well known that the polysaccharide of Ganoderma lucidum exhibits hepatoprotective activity, antitumor activity etc., which many suggest a relationship to lipidperoxidation. The effect of GLP on CCl4- and galactosamineintoxicated cytotoxicity in the primary cultured rat hepatocytes were reduced the GPT value. In order to the estimate the effects of anti-lipidperoxidation of the polysaccharide, enzymatic and nonenzymatic reaction assays were performed, in vitro, in the rat liver microsome. An enzymatic lipidperoxidation reaction by ADP＋FeCl3＋NADPH and CCl4＋NADPH, GLP (1 mg/mL) inhibited 77.4% and 39.4%, respectively, and the nonenzymatic reaction displayed a 97.4% strongly inhibition. In the enzymatic and nonenzymatic inducers treated with GLP, the formation of malondialdehyde (MDA) progressively decreased by raising the GLP concentration. These results suggest that the anti-lipidperoxidation and radical scavenging activity of GLP may play an important part in the liver protection action.

가시오가피의 첨가는 영지버섯 균사체를 이용한 분비 단백질 라카아제 및 셀룰라아제의 생산량을 효율적으로 증가시켰다. 영지버섯 균사체의 생장률 측정에서 가시오가피의 첨가는 영지버섯 균사의 생장률이 톱밥에 비해 절반 수준에 밖에 못미치는 결과를 나타내었다. 반면에 영지버섯 균사의 분비 단백질인 라카아제와 셀룰라아제의 양을 증가시키는데 톱밥에 비해 큰 효과가 있음을 확인할 수 있었다. 가시오가피가 첨가된 배양액에서의 라카아제 활성은 톱밥이 첨가된 배양액에서보다 영지 1호, 영지 2호 및 녹각 영지에서 0.61~2.37배 증가하였고, 이 중에서도 영지 2호는 0.947 U/min으로 라카아제 활성이 가장 높게 평가되었다. 셀룰라아제 활성 평가에서는 가시오가피가 첨가된 배양액에서의 셀룰라아제 활성은 톱밥이 첨가된 배양액에서보다 영지 1호, 영지 2호 및 녹각 영지에서 1.77~2.81배 증가하였고, 이 중에서도 영지 1호 및 영지 2호에 비해 녹각 영지에서 0.172 U/min으로 셀룰라아제 활성이 가장 높게 평가되었다. 따라서 가시오가피의 첨가가 영지버섯 균사의 분비 단백질 생산을 증가시키는데 유용한 소재로 사료된다.

최근 소득이 증가하고 건강에 관심이 증가함에 따라 단순한 미용 화장품보다 여러 가지 기능성을 가진 화장품에 대한 관심이 높아지고 있다. 이에 따라 각종 천연물에 대하여 항산화, 항염 및 미백 등의 활성을 검증하는 다양한 연구가 진행되고 있으며, 본 연구에서도 천연물의 활성과 더불어 다양한 약리작용이 있는 영지버섯 균사체와의 발효에 의한 시너지 효과를 규명하고자 하였다. 인진쑥 영지균사발효 추출물의 DPPH radical 소거 활성은 영지버섯 균사추출물과 유사하게 농도 의존적으로 증가하는 것이 확인되었으나, 저농도의 인진쑥 추출물 보다는 다소 감소하는 것이 확인되었다. 인진쑥 영지균사발효 추출물의 RAW 264.7 세포의 NO 생성 억제활성을 통한 항염활성 평가에 앞서 세포 생존율을 평가한 결과, 흥미롭게도 인진쑥 추출물은 150 μg/mL의 농도부터 RAW 264.7 세포에 대한 생존율을 유의적으로 크게 감소시켰으나, 영지 균사 발효 시 세포 생존율이 크게 증가하는 것이 확인되었다. 영지 균사 발효 시 인진쑥의 독성이 크게 감소하는 것이 확인되었다. 또한 인진쑥 추출물 25 μg/mL의 처리는 동일 농도의 인진쑥 영지버섯 균사발효 추출보다 오히려 유의적인 수준에서 RAW 264.7 세포의 NO생성을 억제하는 것이 확인되었다. 그러나 인진쑥 영지버섯 균사발효는 인진쑥 추출물 자체의 RAW 264.7 세포에 대한 독성을 저감하는 효과가 있어 이로 인해 더욱 높은 농도의 영지버섯 균사발효 추출물 처리를 가능할 것으로 사료된다. 영지 균사 발효에 의한 미백 기능성 소재 발굴이라는 본 연구의 취지에 부합하는 결과가 인진쑥 영지균사발효 추출물 처리구에서 확인되었으며, 200 μg/mL 농도에서는 양성 대조군인 arbutin 보다 약 3.66배의 활성을 보였다. 이러한 결과는 향후 인진쑥 영지균사발효 추출물이 tyrosinase활성을 억제하는 기능성 소재로서의 활용가능성이 크다는 것을 의미하는 것이다.

본 연구에서는 참나무원목을 이용한 영지버섯 재배하우스의 토양의 이화학성조사와 재배지 온도와 같은 영지 관리 특성을 조사하였으며, 초발이 소요일수는 부직포 피복 처리 등 처리 별로 차이 없이 12~13일로 나타났으며, 색 도조사에서 적색도(a)의 경우 비닐처리 19.5, 관행처리 20.2, 부직포처리 19.5로 나타났으며, 황색도(b)는 비닐처리 12.7, 관행처리 13.3, 부직포처리 12.4로 나타나 처리 간 비슷한 경향을 나타내었다. 처리별 생육특성은 관행처리인 무피복 토양의 경우 갓의 길이 145.7 mm, 두께 23.6 mm, 대 길이 73.9 mm, 두께 11.5 mm, 중량은 130 g 을 나타내었고, 부직포피복처리에서는 갓의 길이 157mm, 두께 22.3 mm, 대 길이 93.3 mm, 두께 11.9 mm, 중량은 164 g으로 관행처리보다 우수하였다.

Ganoderma lucidum has been reported to have various biological activities including antioxidant activity. The objective of this study was to compare the antioxidant effects between Ganoderma lucidum (GL) and antler-shaped fruiting body of Ganoderma lucidum (AGL). In vitro antioxidant activities were examined by 2,2-diphenyl-picrylhydrazyl hydrate (DPPH) and 1,2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities of GL and AGL ethanol extracts. In the DPPH and ABTS radical scavenging activities of GL and AGL ethanol extracts, antioxidant activities of AGL extracts (IC50, 66.94 μg/ml and 131.23 μg/ml) was showed higher than GL extracts (IC50, 83.93 μg/ml and 164.54 μg/ml). Total polyphenol content and total flavonoid content (38.00 g GAE/kg extract and 11.58 g NE/kg extract) of AGL were found higher as compared to GL (34.23 g GAE/kg extract and 3.46 g NE/kg extract). In summary, the results of this study demonstrate that AGL extracts had higher antioxidant activities to GL.

The purpose of this study was to investigate whether or not the antler-shaped fruiting body of Ganoderma lucidum (GL) has an anti-inflammatory effect on lipopolysaccharide (LPS)/interferon-γ (IFN-γ)-activated RAW 264.7 macrophage-like cells. To evaluate the anti-inflammatory effects of GL, we examined the inflammatory mediators such as the production of nitric oxide (NO) and the expression of mitogen-activated protein kinase (MAPK), activator protein 1 (AP-1), inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-1β (IL-1β), and interleukin-6 (IL-6). LPS/IFN-γ-induced cellular NO production was significantly decreased in GL-treated RAW 264.7 cells. Moreover, Western blotting analysis results demonstrated that reduced protein expression of MAPK families (such as extracellular signal-regulated kinase, c-Jun amino-terminal kinase, and p38 MAPK) and AP-1-targeting inflammatory enzymes (iNOS, COX-2, IL-1β, and IL-6). These results indicated that GL modulates the MAPK/AP-1 signal pathway in inflammatory process. In conclusion, the present study provides important evidence that GL can potentially be used to reduce LPS/IFN-γ-induced inflammatory response by inhibiting the MAPK/AP-1 signaling pathways.

Ganoderma lucidum is one of the most popular medicinal mushrooms of Asian region having promising immune enhancing properties. This single species of mushroom has over 400 bioactive compounds and possess properties relating to battling tumours, viruses, bacteria, inflammation, platelet-aggregation, immune system modulation among many others. There are documented reports of G. lucidum fighting the activity against HIV/AIDS and diabetes. Owing to such a wide range of medicinal applications, the global consumption of G. lucidum is high, which has resulted in a plethora of commercially available patented products. These products have G. lucidum as an active constituent and are usually marketed as food supplements but are also available in the form of creams, hair tonics and medicinal soaps. The growing demand and the high cost of G. lucidum in the local market have enforced to think about a range of novel methods for the cultivation of this mushroom. We explored different novel strategies in this direction. In the first approach, the widely available agricultural residues of oil palm biomass (empty fruit bunch fibres, EFB) from Malaysia were used to cultivate this mushroom with cheap cost. The required conditions for the growth of this mushroom were studied by employing a pilot-scale cultivation in a mushroom farm. From the results it was scrutinized that oil palm waste fibres are the potential source of cultivating G. lucidum either alone or in combination of substrates. In the second strategy, the effect of UV-C radiation on the physical growth of G. lucidum mycelium has shown positive results. Results of 10 and 15 min of UV-C exposure treatments per 24 h showed a 30% increase in the diameter of mycelium over control in Potato Dextrose Agar medium petri-plates. Using EFB as substrates not only supports the G. lucidum cultivation but also offers a viable alternative for the management of solid waste in oil palm plantations of Malaysia. These results stipulate a better and stronger mushroom in terms of its bioactive compounds content and better growth parameters. Moreover, it also benefits the local mushroom farmers in Malaysia for the easy availability of the substrate materials and with better economy. The techniques involved can be propagated to the local Ganoderma industry for improvised cultivation for better output. A high output would also mean profitable business resulting in revolutionizing the economics of Ganoderma markets all over the world.

In order to determine the possibility of artificial cultivation of Ganoderma lucidum, a study was carried out to optimize the medium composition used for bed culture. Sawdust spawn of medium composition for optimal growth was found to be the combination of 75% oak-sawdust, 10% cotton-seed meal, and 15% beet pulp. The optimal conditions for the formation of fruiting bodies were shown to be a temperature of 28–30oC, light intensity of 1,000–1,500 lux, and CO2 concentration of 1%.

The choice of strain, substrate, and environmental conditions are fundamental factors that determine the ability of a fungus to develop and produce mushrooms with high quality. The objective of this study was to compare the strains of Lentinula edodes and Ganoderma lucidum originated from South Korea and Brazil in relation to their vegetative development in different culture media, to obtain isolates with high quality. The strains of the two mushroom species from Brazil and South Korea grew slowly, possibly because of abiotic factors or storage technique used. However, it may be concluded that temperature, humidity, light, pH, and nutrients from the substrate, if not regulated appropriately may affect the vegetative species.