• Title/Summary/Keyword: fungal cell culture

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Expression of Murine GM-CSF in Recombinant Aspergillus niger

  • Kim, Nyoung-Ji;Kwon, Tae-Ho;Jang, Yong-Suk;Yang, Moon-Sik;Kim, Dae-Hyuk
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.287-292
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    • 2000
  • Recombinant Aspergillus niger was constructed to express and secrete a biologically active murine granulaocyte macrophage-colony stimulating factor (mGM-CSF). A 500 bp fragment encoding the signal peptide and terminator of glyceraldehyde-3-phosphate dehydrogenase (gpd). The hygromycin phosphotrasferase gene (hph) was used as a selection marker for the fungal transformants. An expression vector was introduced into A. niger ATCC 9642, and a Northern blot analysis indicated the presence of a considerable amount of transcripts from the introduced mGM-CSF. The biological activity of recombinant mGM-CSF (rmGM-CSF) isolated from the culture filtrate was confirmend by measuring the proliferationof the GM-CSF dependent FDC-P1 cell line. It appeared that rmGM-CSF was amenable to the proteolytic activity produced by A. niger, since biological actibity was only observed when the transformants were grown in a protease-repressing medium, and the activity of rmGM-CSF dramatically decreased with an increase of age of the culture. The yield of rmGM-CSF, as determined by ELISA. was 640 ng/l of culture filtrate. Accordingly, its specific activity is estimated to be approximately two-and-a-half times higher than that of a commercial preparation from E. coli.

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Antagonistic Activity against Dirty Panicle Rice Fungal Pathogens and Plant Growth-Promoting Activity of Bacillus amyloliquefaciens BAS23

  • Saechow, Sukanya;Thammasittirong, Anon;Kittakoop, Prasat;Prachya, Surasak;Thammasittirong, Sutticha Na-Ranong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1527-1535
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    • 2018
  • Bacterial strain BAS23 was isolated from rice field soil and identified as Bacillus amyloliquefaciens. Based on dual culture method results, the bacterium BAS23 exhibited potent in vitro inhibitory activity on mycelial growth against a broad range of dirty panicle fungal pathogens of rice (Curvularia lunata, Fusarium semitectum and Helminthosporium oryzae). Cell-free culture of BAS23 displayed a significant effect on germ tube elongation and mycelial growth. The highest dry weight reduction (%) values of C. lunata, H. oryzae and F. semitectum were 92.7%, 75.7%, and 68.9%, respectively. Analysis of electrospray ionization-mass spectrometry (ESI-MS) and $^1H$ nuclear magnetic resonance (NMR) spectroscopy revealed that the lipopeptides were iturin A with a C14 side chain (C14 iturinic acid), and a C15 side chain (C15 iturinic acid), which were produced by BAS23 when it was cultured in nutrient broth (NB) for 72 h at $30^{\circ}C$. BAS23, the efficient antagonistic bacterium, also possessed in vitro multiple traits for plant growth promotion and improved rice seedling growth. The results indicated that BAS23 represents a useful option either for biocontrol or as a plant growth-promoting agent.

Effects of Morphology and Rheology on Neo-fructosyltransferase Production by Penicillium citrinum

  • Lim, Jung-Soo;Lee, Jong-Ho;Kim, Jung-Mo;Park, Seung-Won;Kim, Seung-Wook
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.100-104
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    • 2006
  • In this study, we investigated the relationship between the morphology and the rheological properties of Penicillium citrinum to improve the production of neo-fructosyltransferase (neo-FTase). In a 2.5 L bioreactor culture of P. citrinum, it was observed that agitation speed and aeration rate had significant effects on the production of neo-FTase and that maximum cell mass and neo-FTase production obtained at 500 rpm and 1.5vvm were 8.14 g/L and $53.2{\times}10^{-3} U/mL$, respectively. Cell mass and neo-FTase production increased to 91.53 and 25.17%, respectively. In the morphology and rheology studies, P. citrinum showed a typical pellet morphology that was explained by a shaving mechanism; this phenomenon was significantly affected by carbon sources. The rheology of neo-FTase fermentation by P. citrinum was dependent on cell growth and fungal morphology.

Evaluation and Genome Mining of Bacillus stercoris Isolate B.PNR1 as Potential Agent for Fusarium Wilt Control and Growth Promotion of Tomato

  • Rattana Pengproh;Thanwanit Thanyasiriwat;Kusavadee Sangdee;Juthaporn Saengprajak;Praphat Kawicha;Aphidech Sangdee
    • The Plant Pathology Journal
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    • v.39 no.5
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    • pp.430-448
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    • 2023
  • Recently, strategies for controlling Fusarium oxysporum f. sp. lycopersici (Fol), the causal agent of Fusarium wilt of tomato, focus on using effective biocontrol agents. In this study, an analysis of the biocontrol and plant growth promoting (PGP) attributes of 11 isolates of loamy soil Bacillus spp. has been conducted. Among them, the isolates B.PNR1 and B.PNR2 inhibited the mycelial growth of Fol by inducing abnormal fungal cell wall structures and cell wall collapse. Moreover, broad-spectrum activity against four other plant pathogenic fungi, F. oxysporum f. sp. cubense race 1 (Foc), Sclerotium rolfsii, Colletotrichum musae, and C. gloeosporioides were noted for these isolates. These two Bacillus isolates produced indole acetic acid, phosphate solubilization enzymes, and amylolytic and cellulolytic enzymes. In the pot experiment, the culture filtrate from B.PNR1 showed greater inhibition of the fungal pathogens and significantly promoted the growth of tomato plants more than those of the other treatments. Isolate B.PNR1, the best biocontrol and PGP, was identified as Bacillus stercoris by its 16S rRNA gene sequence and whole genome sequencing analysis (WGS). The WGS, through genome mining, confirmed that the B.PNR1 genome contained genes/gene cluster of a nonribosomal peptide synthetase/polyketide synthase, such as fengycin, surfactin, bacillaene, subtilosin A, bacilysin, and bacillibactin, which are involved in antagonistic and PGP activities. Therefore, our finding demonstrates the effectiveness of B. stercoris strain B.PNR1 as an antagonist and for plant growth promotion, highlighting the use of this microorganism as a biocontrol agent against the Fusarium wilt pathogen and PGP abilities in tomatoes.

Analysis of Microflora Profile in Korean Traditional Nuruk

  • Song, Sang Hoon;Lee, Chunghee;Lee, Sulhee;Park, Jung Min;Lee, Hyong-Joo;Bai, Dong-Hoon;Yoon, Sung-Sik;Choi, Jun Bong;Park, Young-Seo
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.40-46
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    • 2013
  • A variety of nuruk were collected from various provinces in Korea, and their microflora profiles were analyzed at the species level. A total of 42 nuruk samples were collected and when the viable cell numbers in these nuruk were enumerated, the average cell numbers of bacteria, fungi, yeast, and lactic acid bacteria from all nuruk were 7.21, 7.91, 3.49, and 4.88 log CFU/10 g, respectively. There were no significant differences in viable cell numbers of bacteria or fungi according to regions collected. Bacillus amyloliquefaciens and B. subtilis were the predominant bacterial strains in most samples. A significant portion, 13 out of 42 nuruk, contained foodborne pathogens such as B. cereus or Cronobacter sakazakii. There were various species of lactic acid bacteria such as Enterococcus faecium and Pediococcus pentosaceus in nuruk. It was unexpectedly found that only 13 among the 42 nuruk samples contained Aspergillus oryzae, the representative saccharifying fungi in makgeolli, whereas a fungi Lichtheimia corymbifera was widely distributed in nuruk. It was also found that Pichia jadinii was the predominant yeast strain in most nuruk, but the representative alcohol fermentation strain, Saccharomyces cerevisiae, was isolated from only 18 out of the 42 nuruk. These results suggested that a variety of species of fungi and yeast were distributed in nuruk and involved in the fermentation of makgeolli. In this study, a total of 64 bacterial species, 39 fugal species, and 15 yeast species were identified from nuruk. Among these strains, 37 bacterial species, 20 fungal species, and 8 yeast species were distributed less than 0.1%.

Penicillium sp.-L4의 균성장 및 효소작용을 억제하는 $\beta$-glucosidase inhibitor의 분리 및 특성

  • Kim, Moo-Sung;Ha, Sung-Yoon;Jeon, Gi-Boong;Lim, Dal-Taek;Park, Byung-Hwa;Lee, Bo-Seop;Lee, Sang-Rin;Choi, Yong-Keel
    • Microbiology and Biotechnology Letters
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    • v.25 no.2
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    • pp.189-196
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    • 1997
  • A producer of inhibitor against ${\beta}-glucosidase$ of Penicillium sp.-L4 was screened from Actinomycetes, and the isolated strain was identified as Streptomyces sp. The inhibitor produced was very stable against heat, acidic and alkaline conditions, proteolytic and amylolytic enzymes. The inhibotor was purified from culture broth through activated carbon treatment, ultrafiltration, anion and cation exchange, activated carbon columm, acetone precipitation and preparative HPLC. It showed inhibitory activities against a variety of dissacharide hydrolyzing enzymes produced by P.sp.-L4, and the mode of inhibition was competitive. Its structure and molecular formular was elucidated by IR, $^1H\;and\;^{13}C$ NMR and FAB/Mass spectrometry, which was identified as 1-deoxynojirimycin (dNM). dNM showed inhibitory effects on the cell growth and hydrolytic enzyme action of P.sp.-L4 on agar plate and infected lemon peel.

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Baicalin Production in Transformed Hairy Root Clones of Scutellaria baicalensis

  • Hwang, Sung-Jin
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.105-109
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    • 2006
  • A transformed hairy root clone of Scutellaria baicalensis was established following infection with Agrobacterium rhizogenes ATCC15834. Three root clones of S baicalensis were selected by growth habit and baicalin content. The most active strain-the SR-03 clone-was examined for its growth and baicalin content under various culture conditions. The root growth and baicalin content were maximized in a Schenk and Hildebrandt medium supplemented with 4 and 6% sucrose, respectively. The accumulation of baicalin in transformed hairy roots was enhanced through exposure to various elicitors. Elicitation was attained by the addition of methyl jasmonate, salicylic acid, and various concentrations of fungal cell wall elicitors to the medium. The accumulation of baicalin in the elicited cultures ranged from 10.5 to 18.3 mg/g dry weight of the roots, which was 1.5- to 3-fold the amount attained in controls.

Taxonomic Characteristics of Six Species of Entomopathogenic Fungi Isolated from the Silkworm, Bombyx mori

  • Nguyen, Mau Tuan;Nam, Sung Hee;Park, Hyun Ro;Han, Myung Sae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.2
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    • pp.229-234
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    • 2004
  • Six entomopathogenic fungus isolates, Beauveria bassiana J57A, Nomuraea rileyi J125A, Paecilomyces farinosus J3A, Paecilomyces fumosoroseus J50A, Metarhizium anisopliae J88, Aspergilius sp. J64A, causing muscardine disease and aspergillosis in the silkworm, Bombyx mori were investigated for their cultural and morphological characteristics (on PDA culture media within 14 days at $24^{\circ}C$). The results showed that they differ each other from the features of cultural characteristics (colony elevation, colony color, colony growth rate) or morphological characteristics (conidiogenous cell structure, phialides, conidia size and shape). Among cultural characteristics, colony color is the easiest recognizable character between isolates. The morphological characteristics of each fungal isolate correspond to the descriptions of current system of classification.

Antiviral, Antimicrobial, and Cytotoxic Properties of Peptavirins A and B Produced by Apiocrea sp.14T

  • Kim, Young-Ho;Yeo, Woon-Hyung;Yun, Bong-Sik;Kim, Young-Sook;Lee, Sang-Jun;Yoo, Ik-Dong;Kim, Kab-Sig;Park, Eun-Kyung;Lee, Jong-Chull
    • The Plant Pathology Journal
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    • v.18 no.1
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    • pp.18-22
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    • 2002
  • Two peptaibol antibiotics, peptavirins A and B, which exhibited strong inhibitory effect against Tobacco mosaic vials (TMV) infection, were isolated from steam-cooked rice culture of Apiocrea sp.14T. The peptavirins were identified as new derivatives of chrysospermins, which are 19-mer and have been reported to be produced in a fungal isolate. The physicochemical properties of the peptavirins were mostly identical with chrysospermins A through D except for the UV absorption spectrum. The peptavirins inhibited the growths of the Grampositive bacteria tested, including the plant pathogenic bacterium, Corynebacterium lilium, and the fungus, Aspergillus niger. Peptavirin A was somewhat cytotoxic to cancer cell lines, especially K562 (leukemia) and UACC 62 (melanoma), whereas peptavirin B only exhibited slight cytotoxicity.

Electron Microscopic Observations of Mouse Liver Cell Treated with Fungal Culture Filtrates Isolated from Foodstuffs II. Results of Isolated Strains (각종 식품에서 분리한 진균 배양액으로 처리된 마우스 간세포의 전자현미경적 관찰 II. 분리균주에 대한 실험성적)

  • Deung, Young-Kun;Choi, Choon-Keun;Koh, Choon-Myung
    • Applied Microscopy
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    • v.3 no.1
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    • pp.45-54
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    • 1973
  • The present study is to determine the toxicity of the fungi isolated from foodstuffs by observing the ultrastructural changes in the mouse liver cells. The results as follows: 1. The toxin-producing fungi were screened by the methods of toxin-screening test(cyto-toxicity test against to HeLa cells and thin layer chromatography). 2. All of the experimental animals treated with isolated fungi were observed the focal necrosis and inflammatory infiltration of liver parenchymal cells. 3. It showed the cytoplasmic changes, such as dilatation of rough endoplasmic reticulum (RER), swelling of mitochondria (mi). increased number of lipid droplet (li) and glycogen (gl), detachment of ribosomes (ri) by observing the electron microscopy. 4. Nuclear and nucleolar alteration were also noted the segregation of nucleolar element and irregularity of nuclear envelopes. 5. As a mass screening, the cytotoxicity test using HeLa cells and thin layer chromatography are feasible methods to detection of the mycotoxin producing fungi from various sources.

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