• Title/Summary/Keyword: Yeast Cell

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A. study on Environmental Tolerance of Yeast S. cevevisiae (효모의 환경내성에 대하여 2)

  • 임억규;김준호
    • Korean Journal of Microbiology
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    • v.16 no.3
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    • pp.103-110
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    • 1978
  • Saccharomyces cerevisiae strain M. was cultured in a molasses-containing media with different amounts of phosphorous and nitrogen sources. The effects of constituents of the cell on the functional activity as well as sensitivity of it were investigated, the results obtained being summarised as follows : Both the thermotolerance and dry tolerance of the yeast cell were higher when the more carbohydrate and thehalose were present in the yeast cell. During the drying, the rate of dead cell was noted increasing and the fermentability decreasing, but it was more remarkable at early stage of the decreasing rate of drying, and at the same time increasing rate of dead cell and decrease of fermentability were more remarkable in the yeast cell containing much protein. In this case the speed of drying was slower. The trehalose content in the yeast cell increased during early stage of the drying and this increase was higher when content of trehalose and carbohydrate in the initial yeast cell was relatively high.

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Yeast two-hybrid assay with fluorescence reporter (형광 리포터를 활용한 효모 단백질 잡종 기법 개발)

  • Park, Seong Kyun;Seo, Su Ryeon;Hwang, Byung Joon
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.199-205
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    • 2019
  • Yeast two-hybrid (Y2H) technique has been used to study protein-protein interactions, but its application particularly to a large-scale analysis of protein interaction networks, is limited by the fact that the technique is labor-intensive, based on scoring colonies on plate. Here, we develop a new reporter for the measurement of the protein-protein interactions by flow cytometry. The yeast harboring interacting proteins can also be enriched by fluorescence-activated cell sorting (FACS) or magnetic-activated cell sorting (MACS). When two interacting proteins are present in the same yeast cell, a reporter protein containing 10 tandem repeats of c-myc epitope becomes localized on the surface of the cell wall, without affecting cell growth. We successful measured the surface display of c-myc epitope upon interacting p53 with SV40 T antigen by flow cytometry. Thus, the newly developed Y2H assay based on the display of c-myc repeat on yeast cell wall could be used to the simultaneous analysis of multiple protein-protein interactions without laborious counting colonies on plate.

Preparation of Corncob Grits as a Carrier for Immobilizing Yeast Cells for Ethanol Production

  • Lee, Sang-Eun;Lee, Choon Geun;Kang, Do Hyung;Lee, Hyeon-Yong;Jung, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1673-1680
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    • 2012
  • In this study, DEAE-corncobs [delignified corncob grits derivatized with 2-(diethylamino)ethyl chloride hydrochloride ($DEAE{\cdot}HCl$)] were prepared as a carrier to immobilize yeast (Saccharomyces cerevisiae) for ethanol production. The immobilized yeast cell reactor produced ethanol under optimized $DEAE{\cdot}HCl$ derivatization and adsorption conditions between yeast cells and the DEAE-corncobs. When delignified corncob grit (3.0 g) was derivatized with 0.5M $DEAE{\cdot}HCl$, the yeast cell suspension ($OD_{600}$ = 3.0) was adsorbed at >90% of the initial cell $OD_{600}$. This amount of adsorbed yeast cells was estimated to be 5.36 mg-dry cells/g-DEAE corncobs. The $Q_{max}$ (the maximum cell adsorption by the carrier) of the DEAE-corncobs was estimated to be 25.1 (mg/g), based on a Languir model biosorption isotherm experiment. When we conducted a batch culture with medium recycling using the immobilized yeast cells, the yeast cells on DEAE-corncobs produced ethanol gradually, according to glucose consumption, without cells detaching from the DEAE-corncobs. We observed under electron microscopy that the yeast cells grew on the surface and in the holes of the DEAE-corncobs. In a future study, DEAE-corncobs and the immobilized yeast cell reactor system will contribute to bioethanol production from biomass hydrolysates.

Comparative Study on Red Blood Cell Hemolysis and Yeast Test by Photosensitizing Compounds (광예민성화합물에 의한 적혈구 용혈현상과 Yeast 시험 비교연구)

  • 김봉희;백광흠
    • Environmental Analysis Health and Toxicology
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    • v.5 no.1_2
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    • pp.45-50
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    • 1990
  • In order to investigate the phototoxicity of five phenothiazine derivatives and one thioxanthen derivative were examined by using in vitro method based on growth inhibition of Candida albicans and red blood cell hemolysis. Effects of the test compounds on RBCs were monitored with a spectrophotometer and a drug PI in the Candida albicans was calculated on the basis of the lowest concentration giving a yeast-free zone. All phenothiazines phototoxic in the red blood cell hemolysis method were positive in the yeast test except promethazine. It was also observed that toxic photo-products were formed by perphenazine and chlorpromazine in the red blood cell hemolysis.

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Property of Yeast Cell Protein from Rice Bran Oil (미강유를 이용한 효모균체 단백질의 특성)

  • 안태영
    • The Korean Journal of Food And Nutrition
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    • v.3 no.1
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    • pp.35-38
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    • 1990
  • For the purpose of the production of single cell protein from rice bran oil, yeast was isolating from soil. It was belonging to Candida albicans Species. These experiments were conducted to find out the property on yeast cell from rice bran oil Molecular weight for the main protein on yeast cell protein from rice bran oil separated by 1% SDS polyacrylamide gel electrophorosis was 22, 000.

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Cultivation of the Hyperthermophilic Archaeon Sulfolobus solfataricus in Low-Salt Media

  • Park, Chan-Beum;Lee, Sun-Bok
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.4 no.1
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    • pp.21-25
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    • 1999
  • Two low-salt complex media, bactopeptone and desalted yeast extract, were used for high density cultivation of the hyperthermophilic archaeon Sulfolobus solfataricus (DSM 1617). Bactopeptone, which has low mineral ion content among various complex media, was good for cell growth in batch cultures; the maximal cell density in bactopeptone was comparable to that in yeast extract. However, cell growth was rather poor when bactopeptone was added by the fed-batch procedure. Since several vitamins are deficient in abctopeptone, the effect of vitamins on cell growth was examined. Among the vitamins tested, pyridoxine was found to improve the growth rate of S. solfataricus. To reduce the growth inhibition caused by mineral ions, yeast extract was dialyzed against distilled water and then fed-batch cultures were carried out using a fed medium containing desalted yeast extract. Although the concentrations of mineral ions in yeast extract were significantly lowered by the dialysis whether low molecular weight solutes in yest extract are crucial for cell growth, we investigated the effect of trehalose, a most abundant compatible solute in yeast extract, on the growth pattern. Cell densities were increased and the length of the lag phase was markedly shortened by the presence of trehalose, indicating that trehalose plays an important role in the growth of S. solfataricus.

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리보핵산 관련물질을 함유한 Yeast Extracts 제조에 Streptomyces faecalis MSF 배양액의 이용

  • 임억규
    • Microbiology and Biotechnology Letters
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    • v.25 no.5
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    • pp.512-519
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    • 1997
  • RNA accumulating strain of Torulopsis versatilis MT-1 was cultured in molasses medium for higher contents of RNA in cell. Yeast cells were harvested at logarithmic phase on synchronous culture. Yield of cells on dry base to input sugar was 59.5%. Crude protein content was 55.1% in cell. RNA content was 13.9%. Some problems found in the process for the preparation of yeast extracts were improved by the addition of culture broth of Streptomyces faecalis MSF which secrete RNase (5' nuclease and 5' adenylic acid deaminase). When the culture broth of S. faecalis MSF was added in autolysis process 46% of RNA in cell was converted to I and G(5' inosinic acid and 5' guanylic acid) in extract. By addition of 3-7% culture broth of S.faecalis MSF in autolysis or enzymolysis process at the start or early stage, RNA in extract was converted easily to I and G and protein in cells was easily extracted and hydrolyzed to amino acid. Taste of those yeast extracts was delicious. The yeasty smell in yeast extracts was removed. And cell debris was easily removed from extract.

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효모 세포벽 분해효소 생산균의 탐색 및 효소생산 최적조건의 조사

  • Cha, Seong-Kwan;Choi, Hea-Suk;Kim, Wang-June;Yoon, Suk-Hoo;Kim, Young-Bae
    • Microbiology and Biotechnology Letters
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    • v.24 no.2
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    • pp.143-148
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    • 1996
  • Thousand actinomycetes and 50 soil samples were used for the isolation of microorganisms producing yeast cell wall lytic enzymes. Among 493 strains producing large clear zones on autolysed washed yeast (AWY), 117 strains were selected on living yeast cell agar plates. With the method of lytic activity, one strain (St-1702) was selected, which was temporarily identified as Streptomyces eurythermus. The optimal condition for enzyme production of this strain was partially determined as follows: incubation of the strain for 3 days at 30$\circ$C in the medium containing 2% freeze dried yeast cell, 1% glucose, 1% K$_{2}$HPO$_{4}$, 0.01% MgSO$_{4}$'7H$_{2}$O, 0.5% peptone, and 0.2% (NH$_{4}$)$_{2}$CO$_{3}$ with pH 7.0. The protoplast formation of yeast by using the enzyme produced by this strain was compared with commercial enzymes.

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Dicyma sp. YCH-37이 생산하는 효모세포벽 용해효소 I. 생산균주의 분리 및 효소의 정제

  • Chung, Hee-Chul;Hahm, Byoung-Kwon;Bai, Dong-Hoon;Hasegawa, Toru;Yu, Ju-Hyun
    • Microbiology and Biotechnology Letters
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    • v.24 no.4
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    • pp.445-451
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    • 1996
  • The strain YCH-37, which produces yeast cell wall lytlc enzyme, was isolated from soil. From the microscopic observation, morphological and cultural characteristics, this strain was identified to fungus, Dicyma sp. So, we named this strain as Dicyma sp. YCH-37. The lytic enzyme effectively lysed Salmonella typhimurium among intact living bacteria and Torulopsis, Hansenula, Zygosaccharomyces among intact living yeast, as well as autoclaved yeast strains. The yeast cell wall lytic enzyme was succesively purified to 204 folds with 13% yields through yeast glucan affinity adsorption and DEAE-cellulose column chromatography. The enzyme was identified to monomeric protein with molecular weight of 25,000 daltons from the results of SDS-PAGE and gel filtration. The optimum pH and temperature for the yeast lytic activity were 8.0 and 50$\circ$C, respectively. The enzyme was stable up to 40$\circ$C, and between pH 4.0-pH 10.0.

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Incorporation of Nasutitermes takasagoensis Endoglucanase into Cell Surface-Displayed Minicellulosomes in Pichia pastoris X33

  • Ou, Jingshen;Cao, Yicheng
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1178-1188
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    • 2014
  • In this study, the yeast Pichia pastoris was genetically modified to assemble minicellulosomes on its cell surface by the heterologous expression of a truncated scaffoldin CipA from Clostridium acetobutylicum. Fluorescence microscopy and western blot analysis confirmed that CipA was targeted to the yeast cell surface and that NtEGD, the Nasutitermes takasagoensis endoglucanase that was fused with dockerin, interacted with CipA on the yeast cell surface, suggesting that the cohesin and dockerin domains and cellulose-binding module of C. acetobutylicum were functional in the yeasts. The enzymatic activities of the cellulases in the minicellulosomes that were displayed on the yeast cell surfaces increased dramatically following interaction with the cohesin-dockerin domains. Additionally, the hydrolysis efficiencies of NtEGD for carboxymethyl cellulose, microcrystal cellulose, and filter paper increased up to 1.4-fold, 2.0-fold, and 3.2-fold, respectively. To the best of our knowledge, this is the first report describing the expression of C. acetobutylicum minicellulosomes in yeast and the incorporation of animal cellulases into cellulosomes. This strategy of heterologous cellulase incorporation lends novel insight into the process of cellulosome assembly. Potentially, the surface display of cellulosomes, such as that reported in this study, may be utilized in the engineering of S. cerevisiae for ethanol production from cellulose and additional future applications.