• Title/Summary/Keyword: yeast form

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Invesigation of Functional Roles of a Protein Kinase in a Fungal Plant Pathogen, Magnaporthe oryzae

  • Han, Joon-Hee;Shin, Jong-Hwan;Kim, Kyoung Su
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.43-43
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    • 2014
  • The rice blast disease caused by of Magnaporthe oryzae is one of the most destructive diseases of rice. By the microarray analysis, we profiled expression changes of genes during conidiation and found out many putative genes that are up-regulated. Among those, we first selected MGG_06399 encoding a dual-specificity tyrosine-regulated protein kinase (DYRK), homologous to YAK1 in yeast. To investigate functional roles of MoYAK1, We made ${\Delta}Moyak1$ mutants by homology dependent gene replacement. The deletion mutant showed a remarkable reduction in conidiation and produced abnormally shaped conidia smaller than those of wild type. The conidia form ${\Delta}Moyak1$ were able to develop a germ tube, but failed to form apppressoria on a hydrophobic coverslip. The ${\Delta}Moyak1$ formed appressria on a hydrophobic cover slip when exogenous cAMP was induced, but the appressoria shape was abnormal. The ${\Delta}Moyak1$ also formed appressoria abberent in shape on onion epidermis and rice sheaths and failed to penetrate the surface of the plants. These data indicate that MoYAK1 is associated with cAMP/PKA pathway and important for conidiation, appressorial formation and pathogenic development in Magnaporthe oryzae. Detailed characterization of MoYAK1 will be presented.

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Removal of Endotoxins and Nucleic Acids Using Submicron-sized Polymeric Particles

  • Kim, Chan Wha;Chokyun Rha
    • Journal of Microbiology and Biotechnology
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    • v.6 no.3
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    • pp.189-193
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    • 1996
  • Submicron-sized polymeric particles (SSPP) were used to remove nucleic acids and endotoxins from cell lysates. The positively charged SSPP selectively adsorb nucleic acids and endotoxins and form complexes with them. The complexes can be easily removed by sedimentation or centrifugation. The removal of nucleic acids and endotoxins using SSPP also can be accomplished in the presence of cell and cell debris. Therefore, nucleic acids and endotoxins can be removed in an initial step of the down-stream processes. In bakers yeast and E. coli lysate systems, the level of DNA could be reduced more than three orders of magnitudes and endotoxins more than seven orders of magnitudes concurrently willi the cell debris removal process using SSPP.

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Some Factors of Effect on Formation of Higher Alcohols during Alcoholic Fermentation in Wine (포도주의 알콜 발효 중 고급 알콜 생성에 영향을 미치는 요인)

  • 최진상;한준표;이용수
    • Food Science and Preservation
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    • v.6 no.1
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    • pp.92-98
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    • 1999
  • Contents of formed higher alcohols did not change about at 20 ppm of total nitrogen, but the contents were most at 200 ppm of total nitrogen especially in iso-amyl alcohol, and the contents showed decrease above at 400 ppm, greatly. Higher alcohols formation were high content at pH 4.0, but the contents were increase according to the condition of glucose and sucrose much. The formation of higher alcohols showed less in fermented condition of no elimination sample than in eliminated a mineral in each. Contents of higher alcohols were less in eliminated sample of biotine and inositol than in control, but the contents were higher than the others, and the contents showed especially high in eliminated nicotinic acid and thiamine. Higher alcohols formation were most at the content of SO\ulcorner in 20 ppm of them. The formation of higher alcohols showed more in Saccharomyces cerevisiae was form more in higher alcohols than Saccharomyces bayanus of two yeast strains.

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MTT 방법에 의한 항진균성 활성효과의 측정

  • Lee, Dong Gun;Lee, Sung Gu;Kim, Kil Lyong;Hahm, Kyung-Soo
    • Microbiology and Biotechnology Letters
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    • v.25 no.3
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    • pp.335-337
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    • 1997
  • In this study, we show a convenient MTT assay for detect the susceptibility of yeast-like form of Trichosporon beigelii against antifungal agents. This assay was developed based on mitocondrial respiration by determining reduction of 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) to formazan. Cells of T beigelii are seeded into 96-well microtiter plates, and antifungal agents, amphotericin B, magainin and CA-ME hybrid peptide were added with various concentrations. After 24 hr incubation, MTT was added, then incubations were continued for 4 hr. Formazan formation was quantified photometrically after extraction of the formazan with acid sodium dodesyl sulfate (SDS). From this assay, we could obtained MICs of antifungal agents against T. beigelii. The presented method can easily be used as an effective methods to assess the antiftingal action of various agents on yeasts with minimal amounts of antifungal agents.

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Hyphal Growth Inhibition by Deer Antler Extract Mimics the Effect of Chitin Synthase Deletion in Candida albicans

  • Park, Hyun-Sook;Jhon, Gil-Ja;Choi, Won-Ja
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.422-425
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    • 1998
  • Chitin synthase null-mutants propagate in yeast form in RPMI medium with suppression of hyphal growth. This hyphal suppression is also observed in the wild type culture grown in RPMI medium supplemented with deer antler extract. To identify the possible target of deer antler extract, the enzymatic activities of chitin synthases were examined. The enzymatic activities of three chitin synthases, CAChsl, CAChs2, and CAChs3, were found to be differentially inhibited by deer antler extract. Of them, CAChsl, was the most sensitive to the extract. These results indicate that deer antler extract causes hyphal suppression, which resembles the effects of chitin synthase deletion, probably through direct inhibition of chitin synthases.

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Recombinant Expression, Isotope Labeling and Purification of the Vitamin D Receptor Binding Peptide

  • Chae, Young-Kee;Singarapu, Kiran;Westler, W. Milo;Markley, John L.
    • Bulletin of the Korean Chemical Society
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    • v.32 no.12
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    • pp.4337-4340
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    • 2011
  • The vitamin D receptor binding peptide, VDRBP, was overexpressed as a fused form with the ubiquitin molecule in Rosetta(DE3)pLysS, a protein production strain of Escherichia coli harboring an induction controller plasmid. The fusion protein was bound to the immobilized metal ions, and the denaturation and renaturation of the fusion protein were performed as a part of the purification procedure. After the elution of the fusion protein, the peptide hormone was released from its fusion partner by using yeast ubiquitin hydrolase (YUH), and subsequently purified by reverse phase chromatography. The purity of the resulting peptide fragment was checked by MALDI-TOF mass and NMR spectroscopy. The final yields of the target peptide were around 5 and 2 mg per liter of LB and minimal media, respectively. The recombinant expression and purification of this peptide will enable structural and functional studies using multidimensional NMR spectroscopy and X-ray crystallography.

Purification of YPTP1 with Immobilized Phosphonomethylphenylalanine-Containing Peptide as an Affinity Ligand

  • Han, Jun-Pil;Kwon, Mi-Yun;Cho, Hyeong-Jin
    • BMB Reports
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    • v.31 no.2
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    • pp.135-138
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    • 1998
  • A previous study on a yeast protein tyrosine phosphatase, YPTP1, using synthetic phosphotyrosine-containing peptides with various sequences as substrates revealed that DADEpYDA exhibits high affinity ($K_m=4{\mu}M$) toward the enzyme. A modified version of this peptide, GDADEpmFDA, immobilized on a resin, was used in this study as an affinity ligand for the purification of YPTP1. Phosphonomethyl-phenylalanine (pmF) was used as a nonhydrolyzable analog of the phosphotyrosine (pY) residue, with properties similar to pY. A protected form of pmF, $Fmoc-pmF(^{t}Bu)_{2}-OH$, was chemically synthesized and introduced during solid-phase peptide sythesis. YPTP1 was onrexpressed in an E. coli strain carrying a plasmid pT7-7-ptpl. Affinity chromatography of the crude lysate afforded PTPI (39 kDa) of about 50% purity.

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use of immobilized cells

  • Jeong, B.C.;Macaskie, L.E.
    • The Microorganisms and Industry
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    • v.19 no.4
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    • pp.2-13
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    • 1993
  • Some microorganisms, including actinomycetes, cyanobacteria, and other bacteria, algae, fungi, and yeast, can accumulate and retain relatively high quantities of heavy metals and radionuclides from their external environments (1-4). Both living and dead cells can be used for biosorptive metal/radionuclide removal from solution. Thus microorganisms and products excreted by or derived from microbial cells (2) may provide an alternative or adjunct to conventional techniuqes of metal removal and recovery. Recent approaches have separated the microbial growth and metal removal process to manipulate production of metal-adsorptive capacity of bacteria and metal removal process. If pre-grown cells are immobilized and used for metal removal, mathematical modeling can be applied to predict immobilized cell reactor behavior under specific process conditions. Waste and microbial adsorbent could be separated from the treated flow in one step. Once treated, the metal waste is concentrated in a small volume of sorbed form for easy metal disposal or recovery.

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생균제로서 Pichia anamala의 pilot scale배양조건 및 동결건조의 최적화

  • Yun, Ji-Yong;Lee, Yong-Ho;Choe, Yu-Jin;Seong, Chang-Geun;Han, Gyu-Beom
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.157-160
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    • 2002
  • Probiotics can form profitab1e microorganisms in human or animal internal organs. When it is fed at domastic animals, can bring the rise of weight and prevention of disease. Pichia anonwla by new probiotics have antibiotic effect and oder reducing effect. we researched about optimal condition of pilot scale fermentation and freezed dry of pichia anonwla.

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High Density Cell Culture of Mucor sp. KCTC 8405P for Production of ${\gamma}-Linolenic$ Acid in Fed-Batch Culture

  • Park, Jong-Hyun;Shin, Hyun-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.1 no.2
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    • pp.126-129
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    • 1991
  • In order to produce ${\gamma}-linolenic$ acid by Mucor sp. KCTC 8405P. the fungus was cultivated in fed-batch culture with two phases. i.e., growth in yeast-like form and induction to hyphal growth by pH shift of the culture medium during cultivation. The synchronous growth of the fungus into the appropriate sizes was important for the high density cell culture of this dimorphic fungus. Dissolved oxygen concentration in the medium did not affect degree of unsaturation of fatty acids and ${\gamma}-linolenic$ acid content. Under the culture conditions applied in this experiment. the fungus was found to produce 100 g/l dry mycelia containing 40% of the lipids, where ${\gamma}-linolenic$ acid comprised about 9% of the total extractable fatty acids.

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