• 미생물학회지
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• 제55권2호
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• pp.157-159
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• 2019

낙동강 주변에서 채취한 토양으로부터 분리한 Pseudomonas fluorescens NBC275 (Pf275) 균주는 식물과 곤충에 병을 일으키는 곰팡이류에 우수한 항균력을 보였다. 본 연구에서는 Pf275 균주의 전체염기서열을 해독하고 분석하였는데, 총 염기서열은 6,610,362 bp였고, GC 함량은 60.9%였다. 염색체는 5,869개의 단백질을 암호화하였고, 16개의 rRNA와 65개의 tRNA로 구성되어 있었다. 유전체의 분석을 통해 항균력을 나타내는 2차 대사산물을 암호화하는 유전자를 확인할 수 있었는데, Pf275 균주는 pyoverdine, 2, 4-diacetylphloroglucinol 및 phenazine 등의 항균물질을 생산하였고, 이들 대사산물에 의해 항균력 및 생물방제효과를 나타내는 것으로 판단된다.

• Journal of Ginseng Research
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• 제45권1호
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• pp.156-162
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• 2021

Background: It is estimated that 20-30% of ginseng crops in Canada are lost to root rot each harvest. This disease is commonly caused by fungal infection with Ilyonectria, previously known as Cylindrocarpon. Previous reports have linked the virulence of fungal disease to the production of siderophores, a class of small-molecule iron chelators. However, these siderophores have not been identified in Ilyonectria. Methods: High-resolution LC-MS/MS was used to screen Ilyonectria and Cylindrocarpon strain extracts for secondary metabolite production. These strains were also tested for their ability to cause root rot in American ginseng and categorized as virulent or avirulent. The differences in detected metabolites between the virulent and avirulent strains were compared with a focus on siderophores. Results: For the first time, a siderophore N,N',N"-triacetylfusarinine C (TAFC) has been identified in Ilyonectria, and it appears to be linked to disease virulence. Siderophore production was suppressed as the concentration of iron increased, which is in agreement with previous reports. Conclusion: The identification of the siderophore produced by Ilyonectria gives us further insight into the root rot disease that heavily affects ginseng crop yields. This research identifies a molecular pathway previously unknown for ginseng root rot and could lead to new disease treatment options.

• Journal of Microbiology and Biotechnology
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• 제33권4호
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• pp.543-551
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• 2023

In this study, five endophytic fungi belonging to the Aspergillus and Alternaria genera were isolated from Lagopsis supina. The antimicrobial activity of all fungal fermented extracts against Staphylococcus and Fusarium graminearum was tested using the cup-plate method. Among them, Aspergillus ochraceus XZC-1 showed the best activity and was subsequently selected for large-scale fermentation and bioactivity-directed separation of the secondary metabolites. Four compounds, including 2-methoxy-6-methyl-1,4-benzoquinone (1), 3,5-dihydroxytoluene (2), oleic acid (3), and penicillic acid (4) were discovered. Here, compounds 1 and 4 displayed anti-fungal activity against F. graminearum, F. oxysporum, F. moniliforme, F. stratum, Botrytis cinerea, Magnaporthe oryzae, and Verticillium dahlia with diverse MIC values (128-512 ㎍/ml), which were close to that of the positive control antifungal, actidione (64-128 ㎍/ml). Additionally, compounds 1 and 4 also exhibited moderate antibacterial activity against S. aureus, Listeria monocytogenes, Escherichia coli, and Salmonella enterica, with low MIC values (8-64 ㎍/ml). Moreover, compounds 1 and 4 displayed selective cytotoxicity against cancer cell lines as compared with the normal fibroblast cells. Therefore, this study proposes that the endophytic fungi from L. supina can potentially produce bioactive molecules to be used as lead compounds in drugs or agricultural antibiotics.

• Journal of Microbiology and Biotechnology
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• 제34권7호
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• pp.1452-1463
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• 2024

Fungi generate different metabolites some of which are intrinsically bioactive and could therefore serve as templates for drug development. In the current study, six endophytic fungi namely Aspergillus flavus, Aspergillus tubigenesis, Aspergillus oryzae, Penicillium oxalicum, Aspergillus niger, and Aspergillus brasiliensis were isolated and identified from the medicinal plant, Silybum marianum. These endophytic fungi were identified through intra transcribed sequence (ITS) gene sequencing. The bioactive potentials of fungal extracts were investigated using several bioassays such as antibacterial activity by well-diffusion, MIC, MBC, anti-biofilm, antioxidant, and haemolysis. The Pseudomonas aeruginosa PAO1 was used to determine the antibiofilm activity. The ethyl acetate extract of Aspergillus flavus showed strong to moderate efficacy against Staphylococcus aureus, Escherichia coli, P. aeruginosa, and Bacillus spizizenii. Aspergillus flavus and Aspergillus brasiliensis exhibited significant antibiofilm activity with IC₅₀ at 4.02 and 3.63 mg/ml, while A. flavus exhibited maximum antioxidant activity of 50.8%. Based on HPLC, LC-MS, and NMR experiments kojic acid (1) and carbamic acid (methylene-4, 1-phenylene) bis-dimethyl ester (2) were identified from A. flavus. Kojic acid exhibited DPPH free radical scavenging activity with an IC₅₀ value of 99.3 ㎍/ml and moderate activity against ovarian teratocarcinoma (CH1), colon carcinoma (SW480), and non-small cell lung cancer (A549) cell lines. These findings suggest that endophytic fungi are able to produce promising bioactive compounds which deserve further investigation.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2018년도 춘계학술대회 및 임시총회
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• pp.13-13
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• 2018

Alternaria is a ubiquitous fungal genus, widely distributed in the environment and a range of different habitats. It includes both plant pathogenic and saprophytic species, which can affect crops in the field or cause post-harvest spoilage of plant fruits and kernels. Numerous Alternaria species cause damage to agricultural products including cereal grains, fruits and vegetables, and are responsible for severe economic losses worldwide. Most Alternaria species have the ability to produce a variety of secondary metabolites, which may play important roles in plant pathology as well as food quality and safety. Alternariol (AOH), alternariol monomethyl ether (AME), tenuazonic acid (TeA), tentoxin (TEN) and altenuene (ALT) are considered the main Alternaria compounds thought to pose a risk to human health. However, food-borne Alternaria species are able to produce many additional metabolites, whose toxicity has been tested incompletely or not tested at all. Both alternariols are mutagenic and their presence in cereal grain has been associated with high levels of human esophageal cancer in China. TeA exerts cytotoxic and phytotoxic properties, and is acutely toxic in different animal species, causing hemorrhages in several organs. The possible involvement of TA in the etiology of onyalai, a human hematological disorder occurring in Africa, has been suggested. Altertoxins (ALXs) have been found to be more potent mutagens and acutely toxic to mice than AOH and AME. Other metabolites, such as TEN, are reported to be phytotoxins, and their toxicity on animals has not been demonstrated up to now. Vegetable foods infected by Alternaria rot are obviously not suitable for consumption. Thus, whole fresh fruits are not believed to contribute significantly with Alternaria toxins to human exposure. However, processed vegetable products may introduce considerable amounts of these toxins to the human diet if decayed or moldy fruit is not removed before processing. The taxonomy of the genus is not well defined yet, which makes it difficult to establish an accurate relationship between the contaminant species and their associated mycotoxins. Great efforts have been made to organize taxa into subgeneric taxonomic levels, especially for the small-spored, food associated species, which are closely related and constitute the most relevant food pathogens from this genus. Several crops of agricultural value are susceptible to infection by different Alternaria species and can contribute to the entry of Alternaria mycotoxins in the food chain. The distribution of Alternaria species was studied in different commodities grown in Argentina. These food populations were characterized through a polyphasic approach, with special interest in their secondary metabolite profiles, to understand their full chemical potential. Alternaria species associated with tomato, bell pepper, blueberry, apples and wheat cultivated in Argentina showed a surprisingly high metabolomic and mycotoxigenic potential. The natural occurrence of Alternaria toxins in these foods was also investigated. The results here presented will provide background for discussion on regulations for Alternaria toxins in foods.

• 한국식품위생안전성학회지
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• 제29권2호
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• pp.85-91
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• 2014

Deoxynivalenol (DON) and related trichothecene mycotoxins are extensively distributed in the cereal-based food and feed stuffs worldwide. Recent climate changes and global grain trade increased chance of exposure to more DON and related toxic metabolites in poorly managed production systems. Monitoring the biological and environmental exposures to the toxins are crucial in protecting human and animals from toxicities of the hazardous contaminants in food or feeds. Exposure biomarkers including urine DON itself are prone to shift to less harmful metabolites by intestinal microbiota and liver metabolic enzymes. De-epoxyfication of DON by gut microbes such as Eubacterium strain BBSH 797 and Eubacterium sp. DSM 11798 leads to more fecal secretion of DOM-1. By contrast, most of plant-derived DON-glucoside is also easily catabolized to free DON by gut microbes, which produces more burden to body. Phase 2 hepatic metabolism also contributes to the glucuronidation of DON, which can be useful urine biomarkers. However, chemical modification could be very typical depending on the anthropologic or genetic background, luminal bacteria, and hepatic metabolic enzyme susceptibility to the toxins in the diet. After toxin exposure, effect biomarkers are also important in estimating the linkage and mechanisms of foodborne diseases in human and animal population. Most prominent adverse effects are demonstrated in the DON-induced immunological and behavioral disorders. For instance, acutely elevated interleukin-8 from insulted gut exposed to dietaty DON is a dominant clinical biomarker in human and animals. Moreover, subchronic exposure to the toxins is associated with high levels of serum IgA, a biological mediator of IgA nephritis. In particular, anorexia monitoring using mouse models are recently developed to monitor the biological activities of DON-induced feed refusal. It is also mechanistically linked to alteration of serotoin and peptide YY, which are promising biomarkers of neurological disorders by the toxins. As animal-alternative biomonitoring, huamn enterocyte-based assay has been developed and more realistic gut mimetic models would be useful in monitoring the effect biomarkers in resposne to toxic contaminants in the future investigations.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2007년도 International Meeting of the Microbiological Society of Korea
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• pp.120-122
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• 2007

All living organisms use numerous signal-transduction pathways to sense and respond to their environments and thereby survive and proliferate in a range of biological niches. Molecular dissection of these signalling networks has increased our understanding of these communication processes and provides a platform for therapeutic intervention when these pathways malfunction in disease states, including infection. Owing to the expanding availability of sequenced genomes, a wealth of genetic and molecular tools and the conservation of signalling networks, members of the fungal kingdom serve as excellent model systems for more complex, multicellular organisms. Here, we employed Cryptococcus neoformans as a model system to understand how fungal-signalling circuits operate at the molecular level to sense and respond to a plethora of environmental stresses, including osmoticshock, UV, high temperature, oxidative stress and toxic drugs/metabolites. The stress-activated p38/Hog1 MAPK pathway is structurally conserved in many organisms as diverse as yeast and mammals, but its regulation is uniquely specialized in a majority of clinical Cryptococcus neoformans serotype A and D strains to control differentiation and virulence factor regulation. C. neoformans Hog1 MAPK is controlled by Pbs2 MAPK kinase (MAPKK). The Pbs2-Hog1 MAPK cascade is controlled by the fungal "two-component" system that is composed of a response regulator, Ssk1, and multiple sensor kinases, including two-component.like (Tco) 1 and Tco2. Tco1 and Tco2 play shared and distinct roles in stress responses and drug sensitivity through the Hog1 MAPK system. Furthermore, each sensor kinase mediates unique cellular functions for virulence and morphological differentiation. We also identified and characterized the Ssk2 MAPKKK upstream of the MAPKK Pbs2 and the MAPK Hog1 in C. neoformans. The SSK2 gene was identified as a potential component responsible for differential Hog1 regulation between the serotype D sibling f1 strains B3501 and B3502 through comparative analysis of their meiotic map with the meiotic segregation of Hog1-dependent sensitivity to the fungicide fludioxonil. Ssk2 is the only polymorphic component in the Hog1 MAPK module, including two coding sequence changes between the SSK2 alleles in B3501 and B3502 strains. To further support this finding, the SSK2 allele exchange completely swapped Hog1-related phenotypes between B3501 and B3502 strains. In the serotype A strain H99, disruption of the SSK2 gene dramatically enhanced capsule biosynthesis and mating efficiency, similar to pbs2 and hog1 mutations. Furthermore, ssk2, pbs2, and hog1 mutants are all hypersensitive to a variety of stresses and completely resistant to fludioxonil. Taken together, these findings indicate that Ssk2 is the critical interface protein connecting the two-component system and the Pbs2-Hog1 pathway in C. neoformans.

• Journal of Microbiology and Biotechnology
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• 제21권1호
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• pp.14-19
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• 2011

A cyclic undecapeptide-family natural product, cyclosporin A (CyA), which is one of the most valuable immunosuppressive drugs, is produced nonribosomally by a multifunctional cyclosporin synthetase enzyme complex in a filamentous fungal strain named Tolypocladium niveum. Previously, structural modifications of cyclosporins such as a regionspecific hydroxylation at the $4^{th}$ N-methyl leucine in a rare actinomycetes called Sebekia benihana were reported to lead to dramatic changes in their bioactive spectra. However, the reason behind this change could not be determined since a system to genetically manipulate S. benihana has not yet been developed. To address this limitation, in this study, we utilized the most commonly practiced gene manipulation techniques including conjugation-based foreign gene transfer-and-expression as well as targeted gene disruption to genetically manipulate S. benihana. Using these optimized genetic manipulation systems, a putative cytochrome P450 hydroxylase (CYP) gene named CYP506, which is involved in CyA hydroxylation in S. benihana, was specifically disrupted and genetically complemented. The S. benihana${\Delta}$CYP506 exhibited a significantly reduced CyA hydroxylation yield as well as considerable yield restoration by functional complementation of the S. benihana CYP506 gene, suggesting that the genetically manipulated S. benihana CYP mutant strains may serve as a more efficient bioconversion host for various valuable metabolites including CyA.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.102.1-102
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• 2003

A bacterial strain YC4963 with antifungal activity against Colletotrichum orbiculare, a causal organism of cucumber anthracnose was isolated from the rhizosphere soil of Siegesbeckia pubescens (Siegesbeckia pubescens Makino；Family：Compositae) in Korea. Based on physiological and biochemical characteristics and 16S ribosomal DNA sequence analysis, the bacterial strain was identified as Pseudomonu aureofaciens. The bacteria also inhibited mycelial growth of several plant fungal pathogens such as Botrytis cinerea, Fusarium oxysporum and Rhizoctonia solani on PDA and 0.1 TSA media. The antibiotic activity was found from the culture filtrate of TSB(tryptic soy broth) and its active compounds were quantitatively bound to XAD adsorber resin. The antibiotic spectrum was broad and growth of C. orbiculare and F. oxysporum, B. cinerea were inhibited at very low concentration. The chemical data from various chromatographic procedures showed that active fraction consisted of at least two phenazine derivatives. However, the metabolites had no inhibitory effect on Pythium ultimum which was reported to be sensitive to phenazine antibiotics. The compounds responsible for the activity are now under investigation.

• 한국식품영양과학회지
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• 제14권3호
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• pp.305-313
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• 1985

There is a trend that the total number of cancer cases is steadily increasing as the population grows. It has been estimated that 85% of the cancer rate in the U.S. is attributed to environmental factors. Among the environmental factors, diet and nutrition appear to be related to the largest number of human cancers. Diet and nutrition might be related to cancer by several mechanisms. Food may contain a direct carcinogen or precursors that become carcinogens by spontanous reactions, or by host metabolism, or through the actions of microbial flora. Chemicals that cause cancers generally have reactive electrophilic centers which can combine with electron-rich atoms in nucleic acids and cause cancers by changing the genetic activity of the cells. A variety of factors in foods might be involved in the etiology of carcinogenesis. Chemicals in food that cause cancers include carcinogens of plants and animal origin and also those in drinking water. Other then these, fungal metabolites alcohol, asbestos, heavy metals, pesticides, and food additives might be included as food carcinogenesis. The method of cooking foods also might contribute to carcinogenesis. Some chemicals in foods act as promoters in carcinogenesis. Prevention of cancers by dietary practises have received much interest. Consumption of certain vegetables or cellulose can reduce carcinogenic activity of several compounds. A variety of antioxidants or micronutrients may be effective anticarciongens. Glutathione in the soluble fraction of the cells, is a major defense against oxidative and alkylating carcinogens. Recently anticarcinogenic activity of chlorophyll was demonstrated. Daily consumption of milk appears to effectively reduce stomach cancer.