• Mycobiology
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• 제51권2호
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• pp.94-108
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• 2023

Termitomyces sp. that grow in symbiosis with fungus-farming Termites have medicinal properties. However, they are rare in nature, and their artificial culture is challenging. The expression of AXL receptor tyrosine kinase and immune checkpoint molecules favor the growth of cancer cells. The study evaluated the optimal conditions for the artificial culture of Termitomyces and their inhibitory activity on AXL and immune checkpoint molecules in lung adenocarcinoma and melanoma cell lines. The culture of 45 strains of Termitomyces was compared. Five strains with marked growth rates were selected. Four of the selected strains form a single cluster by sequence analysis. The mycelium of 4 selected strains produces more fungal mass in potato dextrose broth than in a mixed media. The bark was the most appropriate solid substrate for Termitomyces mycelia culture. The mycelium of all five selected strains showed a higher growth rate under normal CO₂ conditions. The culture broth, methanol, and ethyl acetate of one selected strain (T-120) inhibited the mRNA relative expression of AXL receptor tyrosine kinase and immune checkpoint molecules in cancer cell lines. Overall, these results suggest the potential usefulness of Termitomyces extracts as a coadjuvant therapy in malignant diseases.

• 대한본초학회지
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• 제32권4호
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• pp.33-38
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• 2017

Objectives : Termitomyces albuminosus (Berk.) Heim is one of the famous wild edible mushrooms in the southern part of China. It is known that Termitomyces albuminosus, like Aconitum koreanum used in Korean traditional medicine, contains a kind of cerebroside, termitomycesphin, causing a pharmacologic effect on the neuron system. The pharmacologic effect of Termitomyces albuminosus can be used to possibly replace Aconitum koreanum. However, It needs to be certified as safe before it can be used. Here, a single-oral toxicity test and safety classification was conducted to obtain acute information of the toxicity of dried-Termitomyces albuminosus powder and to secure its safety in clinical applications. Methods : In order to calculate approximate lethal dose(ALD), test substance was orally administered to male and female SD-rat at dose levels of 5,000 and 0 (vehicle control) mg/kg (body weight). Based on the result of this toxicity, also the estimation of safety classification was calculated using the HED-based (human equivalent dose) MOS (margin of safety). Results : There were no mortalities, test substances treatment-related clinical signs, no changes in the body or organ weights, and no gross or histopathological findings at 14 days after treatment with test substance. Thus, the approximate lethal dose of dried-Termitomyces albuminosus powder was considered over 5,000 mg/kg in both female and male mice. Conclusions : Based on the limit dose, 5000 mg/kg, it was estimated that dried-Termitomyces albuminosus powder is classified as "Specified class B" indicating that clinical dose is not limited to patients as safe as food.

• Mycobiology
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• 제51권5호
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• pp.281-287
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• 2023

The symbiotic association between fungus-gardening termites Macrotermes and its fungal symbiont has a moderate degree of specificity-although the symbiotic fungi (Termitomyces) form a monophyletic clade, there is not a one-to-one association between termite species and their fungus-garden associates. Here, we aim to determine the origin and phylogenetic relationships of Termitomyces in Oman. We used sequences of the internal transcribed spacer region (ITS) and the nuclear large subunit ribosomal RNA (LSU rRNA, 25S) gene and analyzed these with sequences of Termitomyces from other geographic areas. We find no evidence for more than a single colonization of Oman by Termitomyces. Unexpectedly, we find Termitomyces in Oman is most closely related to the symbiont of M. subhyalinus in West Africa rather than to those of geographically closer populations in East Africa.

• Mycobiology
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• 제47권4호
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• pp.391-400
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• 2019

A survey of mushrooms was conducted in Xishuangbanna, Yunnan Province, China, in the rainy season (May to October) of 2012, 2013, and 2014, during which 16 specimens of Termitomyces were collected. Preliminary macro- and micro-characteristics, together with ITS sequence data, showed that four of the specimens belonged to a new species (Termitomyces fragilis), while the other 12 belonged to T. aurantiacus, T. eurrhizus, T. globules, T. microcarpus, and T. bulborhizus. In this paper, T. fragilis is introduced as a species new to science based on morphological characterization and phylogenetic analyses. Macro- and micro- morphological descriptions, color photographs and line drawings of the new species, and a phylogenetic tree to show the placement of the new species are provided. T. fragilis is then compared with other closely related taxa in the genus Termitomyces.

• Mycobiology
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• 제48권2호
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• pp.95-103
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• 2020

The genus Termitomyces (Lyophyllaceae, Basidiomycota) is often associated with fungus-feeding termites (Macrotermitinae) due to their strong symbiotic relationships. The genus is widely found exclusively in certain regions of Africa and Asia. They are recognized as edible mushroom within Southeast Asia as well. But it is often misidentified based on morphology by the local communities especially in Malaysia for Chlorophyllum molybdites which is a highly poisonous mushroom. Thus, it is necessary to study the genus for Malaysia with the synergy of using both morphological and molecular identification. In this study, we aim to describe another new species as an addition to the genus Termitomyces found within Sabah, Malaysia. We generated two new sequences (nrLSU and mtSSU) for the new species and a total of 28 nrLSU and mtSSU sequences were retrieved from GenBank for the phylogenetic analysis using maximum likelihood and Bayesian inferences. We identified that the new collection from Sabah province is a new species and named as Termitomyces gilvus based on the termites found in the mound. A phylogeny tree made from the concatenated genes of LSU and mtSSU suggests that T. gilvus is closely related to T. bulborhizus from China. According to our results, the combination of molecular and morphology proved to be a robust approach to re-evaluate the taxonomic status of Termitomyces species in Malaysia. Additional surveys are needed to verify the species diversity and clarify their geographic distribution.

• Journal of Microbiology and Biotechnology
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• 제21권4호
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• pp.412-420
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• 2011

Secretion of cellobiase occurred in a brefeldin A (BFA) uninhibited manner in the filamentous fungus Termitomyces clypeatus. Fluorescence confocal microscopy revealed that application of the drug at a concentration of 50 ${\mu}g$/ml caused arrest of Spitzenkorper assembly at the hyphal tip. This resulted in greater than 30% inhibition of total protein secretion in the culture medium. However, the cellobiase titer increased by 17%, and an additional 13% was localized in the vacuolar fraction en route secretion. The secretory vacuoles formed in the presence of the drug were also found to be bigger (68 nm) than those in the control cultures (40 nm). The enzyme secreted in the presence and absence of BFA revealed a single activity band in both cases in native PAGE and had similar molecular masses (approx. 120 kDa) in SDS-PAGE. The BFA enzyme retained 72% of native glycosylation. It also exhibited a higher stability and retained 98% activity at $50^{\circ}C$, 93.3% activity at pH 9, 63.64% activity in the presence of 1M guanidium hydrochloride, and 50% activity at a glucose concentration of 10 mg/ml in comparison to 68% activity, 75% activity, 36% activity, and 19% activity for the control enzyme, respectively. The observations collectively aimed at the operation of an alternative secretory pathway, distinct from the target of brefeldin A, which bypassed the Golgi apparatus, but still was able to deliver the cargo to the vacuoles for secretion. This can be utilized in selectively enhancing the yield and stability of glycosidases for a successful industrial recipe.

• 현장농수산연구지
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• 제21권1호
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• pp.81-87
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• 2019

흑피흰개미버섯(T. albuminosus) 인공재배를 위한 균사 배양적 특성을 조사한 결과, 원균배지 종류에 따른 흑피흰개미버섯 균사 생장의 영향은 KNCAF 9001와 9002 모두 MEA에서 균사 생장이 89와 81 mm로 가장 좋았고 MEA, YM, PDA 순으로 균사 생장이 좋았다. 배양 온도에 따른 흑피흰개미버섯 균사 생장의 영향은 9001와 9002 모두 89와 77 mm로서 25 ℃에서 균사 생장이 빨랐다. 30 ℃에서 균사 생장이 급감되나 40 ℃에서도 생존하는 광고온성균임을 확인하였다. 배지 pH별로 9001과 9002 모두 89와 88 mm로서 pH 6에서 균사 생장이 빨랐다. 9001의 경우 주재료는 면실피(80%)와 미강(20%) 혼합 시 99 mm/10일로서 가장 좋았으며 콘코브(97 mm), 참나무톱밥(96), 포플러톱밥(93)순으로 좋았다. 영양원은 모든 처리에서 밀기울보다 미강을 처리한 것이 좋았다. 9002의 경우 주재료는 9001과 같은 경향으로 나타났으며 다만, 9001보다 생육이 비교적 늦은 특징이 있었다. 배지의 수분함량에 따른 균사 생장의 영향은 9001, 9002 모두 수분함량이 65%일 때 101, 94 mm/10일로서 가장 좋았다. 수분함량이 65%보다 높거나 낮을 때 균사 생장 속도가 늦어지는 경향이 있었다.

• Mycobiology
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• 제39권2호
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• pp.103-108
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• 2011

Amylases and cellulases are important enzymes that can be utilized for various biological activities. Ten different wild Nigerian mushrooms (Agaricus blazei, Agaricus sp., Corilopsis occidentalis, Coriolus versicolor, Termitomyces clypeatus, Termitomyces globulus, Pleurotus tuber-regium, Podoscypha bolleana, Pogonomyces hydnoides, and Nothopanus hygrophanus) were assayed for production of these secondary metabolites. The results revealed that most of the tested wild fungi demonstrated very good amylase and cellulase activities. With the incorporation of carboxymethyl-cellulose (a carbon source) into the culture medium, Agaricus blazei had the highest amylolytic activity of 0.60 unit/mL (at $25^{\circ}C$, pH 6.8). This was followed in order by P. tuber-regium and Agaricus sp. with 0.42 and 0.39 unit/mL, respectively ($p {\leq} 0.05$). Maltose and sucrose supplementation into the submerged liquid medium made N. hygrophanus and P. hydnoides to exhibit very low amylase activities of 0.09 and 0.11 unit/mL, respectively. Introducing peptone (an organic nitrogen source) into the basal medium enhanced the ability of C. versicolor to produce a cellulase value of 0.74 unit/mL. Other organic nitrogen sources that supported good cellulase activities were yeast extract and urea. Sodium nitrate (inorganic nitrogen source) generally inhibited cellulase production in all mushrooms. The best carbon source was carboxymethyl-cellulose, which promoted very high cellulase activity of 0.67 unit/mL in C. versicolor, which was followed in order by P. tuber-regium, T. chypeatus, and C. occidentalis ($p {\leq} 0.05$). Sucrose was the poorest carbon compound, supporting the lowest values of 0.01, 0.01, and 0.14 unit/mL in P. hydnoides, A. blazei, and Agaricus sp., respectively.

• 한국버섯학회지
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• 제18권1호
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• pp.45-52
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• 2020

계종버섯은 중국에서 맛과 향에서 최고의 버섯으로 평가받고 있으며, 본초강목에 그 효능이 기록되어 전해지고 있으나 효능과 물질연구 및 인공재배법 개발이 요구되고 있는 상황이다. 항산화 활성과 항염증 활성을 가지는 기능성 물질의 확보와 기능성식품 개발을 위한 기초자료를 확보하기 위하여 유기용매 추출물과 열수 추출물을 제조하고, 활성이 높은 분획물을 얻고자 하였다. CH₂Cl₂와 MeOH 혼합용매로 추출한 다음 비극성용매와 극성용매로서 용해성에 따라 분획하고, 열수 추출로 추출물을 확보하였다. 분획물과 추출물을 이용한 항산화 활성 조사에서는 열수 추출물(TA4)의 DPPH 라디칼 소거능에 대한 IC₅₀값이 1.5 mg/mL로 나타났고, MeOH 분획물(TA2)에서는 IC₅₀값이 1.93 mg/mL로 높게 나타났다. NO생성 억제율로 조사한 항염증 활성은 EtOAc 분획물(TA1)은 조추출물이지만 100 ㎍/mL 처리농도에서는 79% NO생성이 억제되고, 200 ㎍/mL 농도에서는 NO가 전혀 생성되지 않았다. TA1 물질을 2차례 TLC로 분리한 TA1-5-6물질은 15 ㎍/mL 처리농도에서 대조구와 비교하여 NO생성이 86% 억제되었고, 30 ㎍/mL 농도에서는 NO 생성이 완전히 억제되었다. 그리고 세포독성 실험에서는 50 ㎍/mL 농도에서도 전혀 독성이 나타나지 않았다. MeOH 분획물(TA2) 유래의 TA2-1-5물질은 30 ㎍/mL 처리농도에서 75% 이상 NO생성이 억제되었으며, 세포 독성은 50 ㎍/mL에서도 아주 낮게 나타났다. 선택적 용매의 선별과 추출 방법의 개발을 통하여 세포 독성이 없고, 생리활성이 뛰어난 계종버섯 분획물 확보가 가능하였고, 기능성식품 및 건강 기능성소재로서의 충분한 잠재력을 확인할 수 있었다.

• 생명과학회지
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• 제26권10호
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• pp.1153-1162
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• 2016

계종버섯(Thermitomyces albuminosus)의 안정성을 입증하기 위하여 사료에 계종버섯의 분말을 5%, 2.5%, 1.25%, 0%를 혼합하여 90일 동안 랫드에 자유 급이 시켜 실험동물의 일반증상, 체중변화, 혈액학적 검사, 부검소견, 임상병리, 조직병리학적 검사의 결과를 토대로 독성을 평가하였다. 독성판정기준은 독성의 정도와 양상에 따라 weight-based classification (독성 강도에 따른 분류)로 분류 하였는데 비시험물질-유래 변화인 NOEL, 시험물질-유래 경미한 변화는 NOAEL, 시험물질-유래 중요한 변화는 LOAEL을 기준으로 나누었다. 시험결과, 수컷 좌, 우측 신장 중량이 용량의존적으로 증가하여 고용량군에서는 12%, 8% 증가 하였다. 하지만 임상, 조직병리학적 결과 독성으로 판단되는 소견이 관찰되지 않아 시험물질-유래 경미한 변화로 판단으로 분류하였다. 따라서 판정기준에 따라 수컷은 NOAEL 암컷은 NOEL이 식이함량 5%로 추정되지만 식약처고시에 따라 암수 모두 NOAEL이 식이함량의 5%로 추정할 수 있다. 따라서 계종버섯은 식품 또는 기능성식품으로의 개발에 문제가 없을 것으로 사료된다.