• Title/Summary/Keyword: 효모 세포

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Breeding of Ethanol-producing and Ethanol-tolerant Saccharomyces cerevisiae using Genome Shuffling (Genome shuffling을 이용한 에탄올 생산 및 내성 효모 균주의 육종)

  • Park, A-Hwang;Kim, Yeon-Hee
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1192-1198
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    • 2013
  • To improve yeast strains for bioethanol production, yeasts with ethanol tolerance, thermotolerance, and ${\beta}$-1,3-glucanase activity were bred using yeast genome shuffling. Saccharomyces cerevisiae $BY4742{\Delta}exg1$/pAInu-exgA, which has extracellular ${\beta}$-1,3-glucanase activity, and the Aspergillus oryzae and S. cerevisiae YKY020 strains, which exhibit ethanol tolerance and thermotolerance, were fused by yeast protoplast fusion. Following cell fusion, four candidate cells (No. 3, 9, 11, and 12 strains) showing thermotolerance at $40^{\circ}C$ were selected, and their ethanol tolerance (7% ethanol concentration) and ${\beta}$-1,3-glucanase activity were subsequently analyzed. All the phenotypes of the two parent cells were simultaneously expressed in one (No. 11) of the four candidate cells, and this strain was called BYK-F11. The BYK-F11 fused cell showed enhanced cell growth, ethanol tolerance, ${\beta}$-1,3-glucanase activity, and ethanol productivity compared with the $BY4742{\Delta}exg1$/pAInu-exgA and YKY020 strains. The results prove that a new yeast strain with different characters and the same mating type can be easily bred by protoplast fusion of yeasts.

Isolation and Identification of Yarrowia lipolytica 504D producing Alkaline Proteinase (Alkaline Proteinase를 생산하는 Yarrowia lipolytica 504D의 분리 동정)

  • Kim, Chang-Hwa;Jin, Ingnyol;Yu, Choon-Bal
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.75-81
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    • 1998
  • The yeast strain 504D, isolated from salted shrimp soup, showed the best proteolytic activity under alkaline condition. The yeast formed vegetative cells in almost optimal media for yeasts, but formed only pseudohyphae in the MM medium containing citric acid and true hyphae in the MM medium containing N-acetylglucosamin and ${\beta}$-D-glucose. The yeast was classified as hemiascomycetes to form ascospores by sexual reproduction, and formed blastospores and athrospores by asexual reproduction. The yeast strain did not assimilate almost of the carbon sources, nitrate and nitrite, but some organic acids and alcohols. The fatty acids of whole cells were composed of 53.67% unsaturated fatty acids and 14.58% saturated, and, especially, C17:1 was observed in this strain but not in two control yeasts. However, almost of all results were very similar to the morphological and physiological characteristics of Yarrowia lipolytica KCCM 12495 and KCCM 35426, except for a little differences which are the composition of fatty acids and the manner of mycellium formation. Therefore, the isolated yeast strain 504D is identified as a Yarrowia lipolytica.

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Cultural Characteristics of a Recombinant Saccharomyces cerevisiae for the Improved Production of a Antibacterial Peptide Defensin of Fleshfly (쉬파리 유래 항균텝티드 Defensin의 생산 증진을 위한 재조합 Saccharomyces cerevisiae의 배양학적 특성)

  • 안종석;강대욱;이준원;김민수;김보연;오원근;민태익
    • Korean Journal of Microbiology
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    • v.36 no.3
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    • pp.236-241
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    • 2000
  • A defensin is an inducible antibacterial peptide from a fleshfly and contains 40 residues basic peptide with six cysteines. For the consiruction of recombinant S cerevisiae expressing defensin, the structural gene coding for active defensin was chemically synthesized and fused in fiam to GAP promoter, MFul preprosequence and the GAL7 transcription terminator, generating a recombinant plasnlid pGMD18. S. ce~evisine 2805 Gells were transror~ned to uracil prototroph by the pGMDl8 arid the transformed cells showing antibacterial activity against 111. luteus TAM1056 were selected by growth inhibition zone assay. The optimal culture conditions for the unprovement of the defensin production of a selected tmdonnant were investigated. The optirmzed medium containing 0.4% yeast extract, 2% corn steep liquor, 2.5% glucose and 0.05% $C_2CO_3$, could be determined and the optimum lemperature. and initial pH could be detennnied as $28^{\circ}C$ and pH 3, ~mpectively. The optimized conditioiis revealed the trvofold Increase in the cell growth and the fourfold in the antibaclerial activity. coinpar-ed with tllc Yl'D medium.

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Construction of spSac3 Null Mutants Defective in mRNA Export (mRNA의 핵에서 세포질로의 이동에 관여하는 spSac3 유전자의 결실돌연변이 제조와 특성 조사)

  • Kang Sook-Hee;Yoon Jin-Ho
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.153-155
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    • 2006
  • We constructed the null mutants of fission yeast Schizosaccharomyces pombe spSac3 gene that is homologous to budding yeast Saccharomyces cerevisiae SAC3 involved in mRNA export out of nucleus. Tetrad analysis showed that the spSac3 is essential for vegetative growth. The spSac3 mutants harboring pREP81X-spSac3 plasmid showed poly(A)+ RNA export defect in the presence of thiamine. These results suggest that spSac3 is also involved in mRNA export from the nucleus.

Cytochemical Observation of Volutin Granules and Activities of Tripolyphosphatase and Polyphosphatase in Saccharomyces uvarum (효모 세포의 Tripolyphosphatase와 Polyphosphatase 활성도 및 Volutin 과립의 세포학적 관찰)

  • Lee, Ki-Sung;Choi, Yong-Keel
    • The Korean Journal of Mycology
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    • v.13 no.3
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    • pp.141-148
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    • 1985
  • To investigate cellular regulation of phosphate metabolism between catabolically repressed and derepressed states in Saccharomyces uvarum, the activities of polyphosphatases, the analysis of polyphosphate and cytochemical observation of volutin granules were examined according to the culture phase and under various phosphate concentrations. As the results, tripolyphosphatase activity was increased more than six-fold during catabolic repression as compared with those of catabolic derepression and the polyphosphatase activity increased at the time of maximal accumulation of acid insoluble polyphosphate 'B'. Of the low molecular weight polyphosphates, tripolyphosphate was mainly detected by thin layer chromatography. When the synthesis of volutin granules in derepressed cells was observed cytochemically, acid insoluble polyphosphate localizing at the cell wall was primarily synthesized and then transferred into the cytoplasm, nucleus and/or vacuole.

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A pattern of cell death induced by 40 kHz ultrasound in yeast cell model (40 kHz 초음파에 의해 유도된 효모세포 모델에서 세포사멸 패턴)

  • Kim, Ji Wook;Kong, Hee Jeong;Kim, Young H.;Kang, Kwang Il
    • The Journal of the Acoustical Society of Korea
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    • v.36 no.3
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    • pp.172-178
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    • 2017
  • Ultrasound has been widely used for biological and medical applications including induction of cell death, but a precise mechanism of induced cell death by ultrasound is controversial. In this study, an irradiation system with 40 kHz ultrasound was developed for a suitable cell death test of a representative unicellular organism, yeast, and used to study the biological effect of ultrasound on inducing cell death. Potassium Iodide (KI) dosimetry was used to devise an optimal system that successfully delivers 40 kHz ultrasound and produces reactive oxygen species in a 1.5 ml Eppendorf tube. Cell death was observed in an ultrasound transmission time-dependent fashion in this system. Thermal effect during irradiation was not observable in ultrasound induced cell death. Co-treatment of 40 kHz ultrasound and hydrogen peroxide showed a synergistic effect in inducing cell death. This finding suggests that 40 kHz ultrasound is related to reactive oxygen species formation. However, NAC (N-acetyl-L-cysteine) oxygen scavenger slightly inhibited the cell death by 40 kHz ultrasound. It was also found that 40 kHz ultrasound induced cell death was slightly inhibited by inhibitors of necrosis or apoptosis (glycyrrhizin or zVAD-fmk). This study suggests that cell death induced by 40 kHz ultrasound may not be exclusively related to reactive oxygen species formation and thermal effects in irradiated yeast cells.

The Distribution of Selenium in Proteins of Saccaromyces Cerevisia and Analysis of Selenomethionine in Specific Protein (효모단백질내의 셀레늄 분포 및특정단백질내의 셀레노메티오닌 분석)

  • Shim Heeyoung Shim;Sangwook Ahn;Yonghyun Ahn
    • Journal of the Korean Chemical Society
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    • v.47 no.4
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    • pp.363-369
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    • 2003
  • Selenized yeast (Se yeast) containing $0.1{\%}$(w/w) of selenium was obtained when the yeast was incubated at a selenium concentration of 1$1.14{\times}10_-3 M$ in rich medium. After washing several times, the inorganic selenium on the cell wall was confirmed with MBRT. There was no indication of inorganic selenium on the cell wall when the blue color in MBRT was stayed for 15 minutes. The selenized yeast was sonicated, then the selenium contained protein was obtained after salting out by ammonium sulfate at the concentration $80{\%}$ saturation. The seven protein bands were seperated by SDS-PAGE and the selenium concentration in protein was measured by ICP-AES. Analytical data showed that the large expressed protein band contained a relatively large amount of selenium. The proteins of the 47kDa was contained the concentrations of 69.5 ${\mu}$ Se/g of most many content. The protein (47 kDa) was seperated from PVDF membrane by tank-electroblotting. The isolated protein was hydrolyzed under acid condition and reacted with PITC. The derivatives of amino acids were analyzed by HPLC and compared with the data obtained from regular yeast. The resulting selenium-yeast was analyzed with the selenomethionine concentration of $2{\%}$ comparaed with general amino acids. The goal of this study is to analyze the selenium concentration in protein bands and measure the degree of biotransformation of selenomethionine in a specific protein.

Screening of Anti-inflammatory Compound-producing Wild Yeasts and Their Microbiological Characteristics (항염증 물질 생산 능력이 우수한 야생효모의 선별 및 이들의 균학적 특성)

  • Bae, Sang-Min;Han, Sang-Min;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.45 no.3
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    • pp.212-223
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    • 2017
  • To screen for potent anti-inflammatory compound-producing yeasts, we evaluated nitric oxide production inhibitory activities in RAW 264.7 macrophage cells using cell-free extracts from 182 non-pathogenic yeasts. Rhodotorula graminis YJ36-1 and Meyerozyma guilliermondii YJ34-2 showed high inhibitory activities of 57.4% and 47.0%, respectively. The microbiological characteristics of these yeasts were investigated. Rhodotorula graminis YJ36-1 formed ascospores and pseudomycelium. This species grew well at $25^{\circ}C$ in yeast extract-peptone-dextrose (YPD) medium, vitamin-free medium, and 5% NaCl-containing YPD medium. Meyerozyma guilliermondii YJ34-2 was an asporogenous yeast and did not form pseudomycelium. This strain also grew well at $30^{\circ}C$ in YPD medium, vitamin-free medium, and 5% NaCl-containing YPD medium.