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Overproduction of Laccase by the White-Rot Fungus Pleurotus ostreatus Using Apple Pomace as Inducer.

Young Jin PARK ; Dae Eun YOON ; Hong Il KIM ; O Chul KWON ; Young Bok YOO ; Won Sik KONG ; Chang Soo LEE

Mycobiology.2014;42(2):193-197.

Laccase activity of Pleurotus ostreatus is significantly increased by the addition of apple pomace. Among various conditions, the best concentration of apple pomace and cultivation time for the production of laccase by P. ostreatus was 2.5% and 9 days, respectively. Reverse transcription polymerase chain reaction analyses of laccase isoenzyme genes, including pox1, pox3, pox4, poxc, poxa3, and poxa1b, revealed a clear effect of apple pomace on transcription induction. Our findings reveal that the use of apple pomace can be a model for the valuable addition of similar wastes and for the development of a solid-state fermenter and commercial production of oyster mushroom P. ostreatus.
Fungi* ; Laccase* ; Pleurotus* ; Polymerase Chain Reaction ; Reverse Transcription

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Neuraminidase Inhibitors from the Fermentation Broth of Phellinus linteus.

Byung Soon HWANG ; Myeong Seok LEE ; Seung Woong LEE ; In Kyoung LEE ; Geon Sik SEO ; Hwa Jung CHOI ; Bong Sik YUN

Mycobiology.2014;42(2):189-192.

During a search for neuraminidase inhibitors derived from medicinal fungi, we found that the fermentation broth of Phellinus linteus exhibited potent neuraminidase inhibitory activity. Through bioassay-guided fractionation, two active compounds were purified from the ethyl acetate-soluble portion of the fermentation broth of P. linteus. These structures were identified as inotilone (1) and 4-(3,4-dihydroxyphenyl)-3-buten-2-one (2) by spectroscopic methods. Compounds 1 and 2 inhibited H1N1 neuraminidase activity with IC50 values of 29.1 and 125.6 microM, respectively, in a dose-dependent manner. They also exhibited an antiviral effect in a viral cytopathic effect reduction assay using MDCK cells. These results suggest that compounds 1 and 2 from the culture broth of P. linteus would be good candidates for the prevention and therapeutic strategies towards viral infections.
Cytopathogenic Effect, Viral ; Fermentation* ; Fungi ; Inhibitory Concentration 50 ; Madin Darby Canine Kidney Cells ; Neuraminidase*

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Lipoxygenase Inhibitory Activity of Korean Indigenous Mushroom Extracts and Isolation of an Active Compound from Phellinus baumii.

Seung Woong LEE ; Ja Gyeong SONG ; Byung Soon HWANG ; Dae Won KIM ; Yoon Ju LEE ; E Eum WOO ; Ji Yul KIM ; In Kyoung LEE ; Bong Sik YUN

Mycobiology.2014;42(2):185-188.

We investigated a total of 335 samples of Korean native mushroom extracts as part of our lipoxygenase (LOX) inhibitor screening program. Among the mushroom-methanolic extracts we investigated, 35 exhibited an inhibitory activity greater than 30% against LOX at a concentration of 100 microg/mL. Especially, Collybia maculata, Tylopilus neofelleus, Strobilomyces confusus, Phellinus gilvus, P. linteus, P. baumii, and Inonotus mikadoi exhibited relatively potent LOX inhibitory activities of 73.3%, 51.6%, 52.4%, 66.7%, 59.5%, 100.0%, and 85.2%, respectively. Bioassay-guided fractionation led to the isolation of inoscavin A from the methanolic extract of P. baumii, which showed the most potent activity and was identified by spectroscopic methods. Specifically, inoscavin A exhibited potent LOX inhibitory activity with an IC50 value of 6.8 microM.
Agaricales* ; Inhibitory Concentration 50 ; Lipoxygenase* ; Mass Screening ; Methanol

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Isolation and Analysis of the Enzymatic Properties of Thermophilic Fungi from Compost.

Hanbyul LEE ; Young Min LEE ; Yeongseon JANG ; Sangjoon LEE ; Hwanhwi LEE ; Byoung Jun AHN ; Gyu Hyeok KIM ; Jae Jin KIM

Mycobiology.2014;42(2):181-184.

To the best of our knowledge, this is the first report on thermophilic fungi isolated in Korea. Three species of thermophiles were isolated from compost and were identified as Myriococcum thermophilum, Thermoascus aurantiacus, and Thermomyces lanuginosus. They can grow at temperatures above 50degrees C and produce high levels of cellulolytic and xylanolytic enzymes at high temperatures. Notably, the considerable thermostability of the endo-glucanase produced by T. aurantiacus has made the fungus an attractive source of industrial enzymes.
Cellulase ; Fungi* ; Korea ; Soil* ; Thermoascus

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Genetic Diversity and Pathogenicity of Cylindrocarpon destructans Isolates Obtained from Korean Panax ginseng.

Jeong Young SONG ; Mun Won SEO ; Sun Ick KIM ; Myeong Hyeon NAM ; Hyoun Sub LIM ; Hong Gi KIM

Mycobiology.2014;42(2):174-180.

We analyzed the genetic diversity of Cylindrocarpon destructans isolates obtained from Korean ginseng (i.e., Panax ginseng) roots by performing virulence tests and nuclear ribosomal gene internal transcribed spacer (ITS) and mitochondrial small subunit (mt SSU) rDNA sequence analysis. The phylogenetic relationship analysis performed using ITS DNA sequences and isolates from other hosts helped confirm that all the Korean C. destructans isolates belonged to Nectria/Neonectria radicicola complex. The results of in vivo and ex vivo virulence tests showed that the C. destructans isolates could be divided into two groups according to their distinctive difference in virulence and the genetic diversity. The highly virulent Korean isolates in pathogenicity group II (PG II), together with foreign isolates from P. ginseng and P. quinquefolius, formed a single group. The weakly virulent isolates in pathogenicity group I, together with the foreign isolates from other host plants, formed another group and exhibited a greater genetic diversity than the isolates of PG II, as confirmed by the mt SSU rDNA sequence analysis. In addition, as the weakly virulent Korean isolates were genetically very similar to the foreign isolates from other hosts, they were likely to originate from hosts other than the ginseng plants.
Base Sequence ; DNA, Ribosomal ; Genetic Variation* ; Panax* ; Sequence Analysis ; Virulence*

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Cloning and Molecular Characterization of beta-1,3-Glucan Synthase from Sparassis crispa.

Yun Hui YANG ; Hyeon Woo KANG ; Hyeon Su RO

Mycobiology.2014;42(2):167-173.

A beta-glucan synthase gene was isolated from the genomic DNA of polypore mushroom Sparassis crispa, which reportedly produces unusually high amount of soluble beta-1,3-glucan (beta-glucan). Sequencing and subsequent open reading frame analysis of the isolated gene revealed that the gene (5,502 bp) consisted of 10 exons separated by nine introns. The predicted mRNA encoded a beta-glucan synthase protein, consisting of 1,576 amino acid residues. Comparison of the predicted protein sequence with multiple fungal beta-glucan synthases estimated that the isolated gene contained a complete N-terminus but was lacking approximately 70 amino acid residues in the C-terminus. Fungal beta-glucan synthases are integral membrane proteins, containing the two catalytic and two transmembrane domains. The lacking C-terminal part of S. crispa beta-glucan synthase was estimated to include catalytically insignificant transmembrane alpha-helices and loops. Sequence analysis of 101 fungal beta-glucan synthases, obtained from public databases, revealed that the beta-glucan synthases with various fungal origins were categorized into corresponding fungal groups in the classification system. Interestingly, mushrooms belonging to the class Agaricomycetes were found to contain two distinct types (Type I and II) of beta-glucan synthases with the type-specific sequence signatures in the loop regions. S. crispa beta-glucan synthase in this study belonged to Type II family, meaning Type I beta-glucan synthase is expected to be discovered in S. crispa. The high productivity of soluble beta-glucan was not explained but detailed biochemical studies on the catalytic loop domain in the S. crispa beta-glucan synthase will provide better explanations.
Agaricales ; Cell Wall ; Classification ; Clone Cells* ; Cloning, Organism* ; DNA ; Efficiency ; Exons ; Glycogen Synthase ; Humans ; Introns ; Membrane Proteins ; Open Reading Frames ; RNA, Messenger ; Sequence Analysis

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Biocontrol of Fusarium Crown and Root Rot and Promotion of Growth of Tomato by Paenibacillus Strains Isolated from Soil.

Sheng Jun XU ; Byung Sup KIM

Mycobiology.2014;42(2):158-166.

In this study, bacterial strains were isolated from soils from 30 locations of Samcheok, Gangwon province. Of the isolated strains, seven showed potential plant growth promoting and antagonistic activities. Based on cultural and morphological characterization, and 16S rRNA gene sequencing, these strains were identified as Paenibacillus species. All seven strains produced ammonia, cellulase, hydrocyanic acid, indole-3-acetic acid, protease, phosphatase, and siderophores. They also inhibited the mycelial growth of Fusarium oxysporum f. sp. radicis-lycopersici in vitro. The seven Paenibacillus strains enhanced a range of growth parameters in tomato plants under greenhouse conditions, in comparison with non-inoculated control plants. Notably, treatment of tomato plants with one identified strain, P. polymyxa SC09-21, resulted in 80.0% suppression of fusarium crown and root rot under greenhouse conditions. The plant growth promoting and antifungal activity of P. polymyxa SC09-21 identified in this study highlight its potential suitability as a bioinoculant.
Ammonia ; Cellulase ; Crowns* ; Fusarium* ; Gangwon-do ; Genes, rRNA ; Hydrogen Cyanide ; Lycopersicon esculentum* ; Paenibacillus* ; Plants ; Plasmodiophorida ; Siderophores ; Soil*

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A Ferroxidase, Cfo1, Regulates Diverse Environmental Stress Responses of Cryptococcus neoformans through the HOG Pathway.

Kyung Tae LEE ; Jang Won LEE ; Dohyun LEE ; Won Hee JUNG ; Yong Sun BAHN

Mycobiology.2014;42(2):152-157.

The iron uptake and utilization pathways play a critical role in allowing human pathogens, including Cryptococcus neoformans, the causative agent of fatal meningoencephalitis, to survive within the mammalian body by competing with the host for iron. Here we show that the iron regulon is also required for diverse environmental stress responses and that in C. neoformans, it is regulated by the high-osmolarity glycerol response (HOG) pathway. Between CFO1 and CFO2, two ferroxidase genes in the iron regulon, CFO1 but not CFO2 was induced during oxidative and osmotic stress. Interestingly, we found that the HOG pathway repressed basal expression of both CFO1 and CFO2. Furthermore, when the HOG pathway was blocked, CFO2 also responded to oxidative and osmotic stress and the response of CFO1 was increased. We also established that CFO1 plays a major role in responding and adapting to diverse environmental stresses, including oxidative and genotoxic damage, osmotic fluctuations, heavy metal stress, and stress induced by cell membrane destabilizers. Therefore, our findings indicate that in C. neoformans, the iron uptake and utilization pathways are not only required for iron acquisition and survival, but also play a significant role in the environmental stress response through crosstalk with the HOG pathway.
Cell Membrane ; Ceruloplasmin* ; Cryptococcus neoformans* ; Glycerol ; Humans ; Iron ; Meningoencephalitis ; Osmotic Pressure ; Regulon

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Diversity of Endophytic Fungi Isolated from Korean Ginseng Leaves.

Ju Kyeong EO ; Min Seok CHOI ; Ahn Heum EOM

Mycobiology.2014;42(2):147-151.

We investigated the diversity of the foliar endophytes of Korean ginseng. Endophytic fungi were isolated from healthy leaves of mountain-cultivated ginseng (MCG) and field-cultivated ginseng (FCG) at 4 sites in Chungbuk Province. A total of 24 species of fungal endophytes were identified using molecular approaches. Additionally, the diversity of these endophytic fungi was compared between MCG and FCG. The major isolated endophytes were Edenia gomezpompae and Gibberella moniliformis in the MCG and FCG samples, respectively. The results suggest that ginseng endophytes have different community structures in different environments, and this understanding may prove useful in ginseng cultivation.
Chungcheongbuk-do ; Endophytes ; Fungi* ; Gibberella ; Moniliformis ; Panax*

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Wood Decay Fungi in South Korea: Polypores from Seoul.

Yeongseon JANG ; Seokyoon JANG ; Jaejung LEE ; Hwanhwi LEE ; Hanbyul LEE ; Young Min LEE ; Joo Hyun HONG ; Mihee MIN ; Young Woon LIM ; Changmu KIM ; Jae Jin KIM

Mycobiology.2014;42(2):140-146.

In Seoul, a majority of plant communities have undergone significant changes over the last few decades; however, how wood decay fungi have responded and adapted to the changes in vegetation remains unknown. Through an ongoing investigation of Korean indigenous fungi, ca. 300 specimens with poroid basidiocarp were collected in Seoul during 2008~2012. Morphological examination and molecular analysis using the internal transcribed spacer and nuclear large subunit ribosomal DNA region sequences helped identify 38 species belonging to 28 genera, 10 families, and 5 orders in this area. Among them, three polypores, Abundisporus pubertatis, Coriolopsis strumosa, and Perenniporia maackiae were found to be new to South Korea.
Basidiomycota ; Classification ; DNA, Ribosomal ; Fruiting Bodies, Fungal ; Fungi* ; Humans ; Korea ; Maackia ; Phylogeny ; Plants ; Seoul ; Wood*

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