• Title/Summary/Keyword: S. cerevisiae cells

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Assessment of Ethanol Fermentation with Rice Bran by Yeasts (米糠배지에서 酵母에 의한 에탄올 발효액의 평가)

  • 손경현;윤종수;성용분;이강표;김재철;이재흥
    • Microbiology and Biotechnology Letters
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    • v.20 no.1
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    • pp.85-90
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    • 1992
  • Rice bran was employed as a main medium component for ethanol fermentation by Saccharomyces species. Among the several strains of .Saccharomyces yeasts. S. cerevisiae IF0 2346 was selected as the hest strain in view of the interest in the production of ethanol and amino acids. It was found that S. cerevisiae IF0 2346 showed $3\times 10^8$cells/d and 4.7% (v/v) ethanol production after 72 hr cultivation. Although total amount of free amino acids was decreased from 1.099 mg/l to 829 mg/l during the fermentation, glutamic acid. histidine, and isoleucine were increased considerably. With the supplement of 5% glucose to the ferrnentation medium, both ethanol and amino acid production were increased up to 134% and 264%, respectively. compared to the control case. Glutamic. acid, leucine, alanine. phenylalanint:, and valine were the major amino acids in the fermentation broth.

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Role of Osmotic and Salt Stress in the Expression of Erythrose Reductase in Candida magnoliae

  • Park, Eun-Hee;Lee, Ha-Yeon;Ryu, Yeon-Woo;Seo, Jin-Ho;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1064-1068
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    • 2011
  • The osmotolerant yeast, Candida magnoliae, which was isolated from honeycomb, produces erythritol from sugars such as fructose, glucose, and sucrose. Erythrose reductase in C. magnoliae (CmER) reduces erythrose to erythritol with concomitant oxidation of NAD(P)H. Sequence analysis of the 5'-flanking region of the CmER gene indicated that one putative stress response element (STRE, 5'-AGGGG-3'), found in Saccharomyces cerevisiae, exists 72 nucleotides upstream of the translation initiation codon. An enzyme activity assay and semiquantitative reverse transcription polymerase chain reaction revealed that the expression of CmER is upregulated under osmotic and salt stress conditions caused by a high concentration of sugar, KCl, and NaCl. However, CmER was not affected by osmotic and oxidative stress induced by sorbitol and $H_2O_2$, respectively. The basal transcript level of CmER in the presence of sucrose was higher than that in cells treated with fructose and glucose, indicating that the response of CmER to sugar stress is different from that of GRE3 in S. cerevisiae, which expresses aldose reductase in a sugarindependent manner. It was concluded that regulation of CmER differs from that of other aldose reductases in S. cerevisiae.

Biomass Production of Saccharomyces cerevisiae KFCC 10823 and Its Use in Preparation of Doenjang

  • Yoo, Jin-Young;Kim, Hyeon-Gyu;Kwon, Dong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.7 no.1
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    • pp.75-80
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    • 1997
  • An ethanolic fermentation process was developed for preparing Doenjang with high ethanol. Higher and efficient viable cell production of salt-tolerant ethanolic yeast is a prerequisite for the successful commercial-scale process of ethanol production during Doenjang fermentation. Culture conditions of salt-tolerant yeast, S. cerevisiae KFCC 10823, was studied in terms of the effect of several environmental and nutritional factors. Viable cell numbers were the highest in a medium containing the following components per liter of water: soysauce, 300ml; dextrose, 50 g; beef extract, 5 g; yeast extract, 5 g; $KH_2PO_4$, 5 g; NaCl, 50 g. The optimal culture conditions of S. cerevisiae KFCC 10823 were pH 5.5, $25^{\circ}C$, 200 rpm and 0.5 vvm. Yeast viability during batch fermentation was gradually decreased to a level less than $90{\%}$ after 35 hours. The maximum cell number was $2.2{\times}10_7$ cells/ml at the optimal condition. Doenjang prepared with ethanolic yeast was ripened after 45 days at $30^{\circ}C$. This Doenjang contains 470 mg% of amino nitrogen and 2.5% ethanol. The shelf-life at $30^{\circ}C$ was theoretically estimated as 444 days.

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Photosterilization effects of microbial cells by titanium oxide catalyzer ($TiO_2$ 촉매를 이용한 광살균 효과)

  • Joo, Hyun-Kyu
    • Applied Biological Chemistry
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    • v.35 no.4
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    • pp.294-299
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    • 1992
  • In order to examine the effect of $TiO_2$ on photosterilization of microorganism. Proper content of $TiO_2$, illumination time, wave length specificity and cell concentration were investigated for photosterilization of E. coli, B. subtilis and S. cerevisiae. The amount of $TiO_2$ for photosterilization of E. coli was effective in the range of $5{\sim}20\;mg%$. The sterilization time was reduced when the low wave length below 400 nth was not cut off by glass, but the catarizing effect of $TiO_2$ was similar to overall wave length. The photosterilization effects by $TiO_2$ addition was recognized among S. cerevisiae, E. coli and B. subtilis in that order. The photosterilization time of S. cerevisiae was considerably reduced at $10^4/ml$ cell concentration compared to $10^5/ml$.

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Sterilization of Bacteria, Yeast, and Bacterial Endospores by Atmospheric-Pressure Cold Plasma using Helium and Oxygen

  • Lee Kye-Nam;Paek Kwang-Hyun;Ju Won-Tae;Lee Yeon-Hee
    • Journal of Microbiology
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    • v.44 no.3
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    • pp.269-275
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    • 2006
  • Atmospheric-pressure cold plasma (APCP) using helium/oxygen was developed and tested as a suitable sterilization method in a clinical environment. The sterilizing effect of this method is not due to UV light, which is known to be the major sterilization factor of APCP, but instead results from the action of reactive oxygen radicals. Escherichia coli, Staphylococcus aureus, and Saccharomyces cerevisiae deposited on a nitrocellulose filter membrane or Bacillus subtilis spores deposited on polypropylene plates were exposed to helium/oxygen plasma generated with AC input power at 10 kHz, 6 kV. After Plasma treatment, nitrocellulose filter membranes were overlaid on fresh solid media and CFUs were counted after incubation overnight. D-values were 18 sec for E. coli, 19 sec for S. aureus, 1 min 55 sec for S. cerevisiae, and 14 min for B. subtilis spores. D-values of bacteria and yeast were dependent on the initial inoculation concentration, while the D-value of B. subtilis spores showed no correlation. When treated cells were observed with a scanning electron microscope, E. coli was more heavily damaged than S. aureus, S. cevevisiae exhibited peeling, and B. subtilis spores exhibited shrunken morphology. Results showed that APCP using helium/oxygen has many advantages as a sterilization method, especially in a clinical environment with conditions such as stable temperature, unlimited sample size, and no harmful gas production.

Characterization of RAD3 Homologous Gene from Coprinus cinereus (균류 Coprinus cinereus에서 DNA 회복에 관여하는 RAD3 유사유전자의 분리와 특성)

  • Choi In Soon
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.1023-1027
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    • 2004
  • The RAD3 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. An yeast RAD3 gene has been previously isolated by functional complementation. In order to identify the RAD3 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD3 DNA, and then isolated RAD3 homologous DNA from C. cinereus chromosome. The RAD3 homolog DNA was contained in 3.2 kb DNA fragment. Here, we report the results of characterization of a fungus C. cinereus homolog to the yeast RAD3 gene. Southern blot analysis confirmed that the C. cinereus chromosome contains the RAD3 homolog gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from the C. cinereus cells were hybridized with the 3.4 kb PvuII DNA fragment of the S. cerevisiae RAD3 gene, transcripts size of 2.8 kb were detected. In order to investigate whether the increase of the amount of transcripts by DNA damaging agent, transcript levels were examined after treating agents to the cells. The level of transcripts were not increased by untraviolet light (UV). This result indicated that the RAD3 homologous gene is not UV inducible gene. Gene deletion experiments indicate that the HRD3 gene is essential for viability of the cells and DNA repair function. These observations suggest an evolutionary conservation of other protein components with which HRD3 interacts in mediating its DNA repair and viability functions.

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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Analysis for Regulatory Elements in Yeast MGMT Gene Transcription

  • Joo, Jae-Hoon;Kim, Woo-Jae;Rho, Jae-Kyun;Choe, Jae-Hyun;Choe, Soo-Young;Sang-Dai
    • Animal cells and systems
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    • v.2 no.2
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    • pp.287-295
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    • 1998
  • The Saccharomyces cerevisiae MGMT gene encodes a O6-methylguanine DNA methyltransferase that protects cells from mutation or death by DNA alkylating agents. Using an in vitro transcription system, we analyzed its promoter region to find regulatory elements for transcription initiation. DNase I footprinting and a transcription assay showed that a functional TATA box, 5'-TGATATAGCA-3', is located in the region spanning from -25 to -34. We also found one upstream repressing sequence (URS), -333 to -213, by promoter deletion and competition analysis. Gel mobility shift assays and Southwestern blot analysis using URS region indicate specific complex formations. These results indicate that several cis-acting and trans-acting elements might be involved in the transcriptional regulation of the S. cerevisiae MGMT gene.

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Effect of Pulsed Electric Fields upon Accumulation of Zinc in Saccharomyces cerevisiae

  • Pankiewicz, Urszula;Jamroz, Jerzy
    • Journal of Microbiology and Biotechnology
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    • v.21 no.6
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    • pp.646-651
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    • 2011
  • Cultures of Saccharomyces cerevisiae were treated with pulsed electric fields to improve accumulation of zinc in the biomass. Under optimized conditions, that is, on 15 min exposure of the 20 h grown culture to PEFs of 1500 V and 10 ${\mu}s$ pulse width, accumulation of zinc in the yeast biomass reached a maximum of 15.57 mg/g d.m. Under optimum zinc concentration (100 ${\mu}g$/ml nutrient medium), its accumulation in the cells was higher by 63% in comparison with the control (without PEFs). That accumulation significantly correlated against zinc concentration in the medium. Neither multiple exposure of the cultures to PEFs nor intermittent supplementation of the cultures with zinc increased the zinc accumulation. The intermittent supplementation of the cultures with zinc and multiple exposures on PEFs could even reduce the accumulation efficiency, respectively, by 57% and 47%.

Production of Cellulosic Ethanol in Saccharomyces cerevisiae Heterologous Expressing Clostridium thermocellum Endoglucanase and Saccharomycopsis fibuligera β-glucosidase Genes

  • Jeon, Eugene;Hyeon, Jeong-eun;Suh, Dong Jin;Suh, Young-Woong;Kim, Seoung Wook;Song, Kwang Ho;Han, Sung Ok
    • Molecules and Cells
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    • v.28 no.4
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    • pp.369-373
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    • 2009
  • Heterologous secretory expression of endoglucanase E (Clostridium thermocellum) and ${\beta}$-glucosidase 1 (Saccharomycopsis fibuligera) was achieved in Saccharomyces cerevisiae fermentation cultures as an ${\alpha}$-mating factor signal peptide fusion, based on the native enzyme coding sequence. Ethanol production depends on simultaneous saccharification of cellulose to glucose and fermentation of glucose to ethanol by a recombinant yeast strain as a microbial biocatalyst. Recombinant yeast strain expressing endoglucanase and ${\beta}$-glucosidase was able to produce ethanol from ${\beta}$-glucan, CMC and acid swollen cellulose. This indicates that the resultant yeast strain of this study acts efficiently as a whole cell biocatalyst.