• Title/Summary/Keyword: natural yeast

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Construction of the recombinant yeast strain with transformation of rice starch-saccharification enzymes and its alcohol fermentation (유전자 형질전환을 통한 쌀 전분 분해효소 재조합 효모균주의 개발과 발효특성조사)

  • Lee, Ja-Yeon;Chin, Jong-Eon;Bai, Suk
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.220-225
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    • 2016
  • To improve antioxidant glutathione (GSH) content and saccharification ability in sake yeasts of Saccharomyces cerevisiae, the ${\gamma}$-glutamylcysteine synthetase gene (GSH1) from S. cerevisiae, glucoamylase gene (GAM1) and ${\alpha}$-amylase gene (AMY) from Debaryomyces occidentalis were co-expressed in sake yeasts for manufacturing a refreshing alcoholic beverage abundant in GSH from rice starch. The extracellular GSH content of the recombinant sake yeasts increased 1.5-fold relative to the parental wide-type strain. The saccharification ability by glucoamylase of the new yeast strain expressing both GAM1 and AMY genes was 2-fold higher than that of the yeast strain expressing only GAM1 gene when grown in the culture medium containing 2% (w/v) rice starch. It generated 11% (v/v) ethanol from 20% (w/v) rice starch and consumed up to 90% of the starch content after 7 days of fermentation.

Development of Recombinant Human Growth Hormone in Yeast: Efficacy Evaluation and Safety Assessment (Human growth hormone의 개발과 이에 따른 효능 및 안전성 평가)

  • Lee Sangkyun;Park Soon Jae
    • Proceedings of the Korean Society of Toxicology Conference
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    • 1997.10a
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    • pp.38-42
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    • 1997
  • Human growth hormone is known as one of the peptide hormones which is consisted of 191 amino acids derived from the pituitary gland in humans. The objectives of this study were to supply inexpensive recombinant methionyl human growth hormones (rHGH) synthesized by the DNA technology in a yeast cell line and followed by the establishement of protein purification techniques. The next steps of the research were to study its physic-chemical properties and biological properties, and to evaluate various preclinical aspcts including pharmacokinetics sutdy, general pharmacology study, general toxicity test, and specific toxicity tests. Clinical phase I, II, III studies were also done against growth hormone dficient children to reveal that growth promoting effects were similar compared with the natural HGH extracted from pituitary glands and commercially available rHGHs. The results could be summarized that (I) this yeast dervied rHGH have had excellent physico-chemical and biological properties in comparison with a natural HGH and other synthesized rHGHs, (2) we could not see any toxic side effects when very high doses were administered to the experimental animals, and (3) this growth hormone showed effectiveness in the growth stimulating to growth hormone deficient patients.

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Quality Characteristics of Rye Mixed Bread Prepared with Substitutions of Naturally Fermented Raisin Extract and Sourdough (건포도 천연 발효액과 Sourdough를 이용한 호밀 혼합빵의 품질 특성)

  • Kim, Mun-Yong;Chun, Soon-Sil
    • Journal of the East Asian Society of Dietary Life
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    • v.18 no.1
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    • pp.87-94
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    • 2008
  • In this study, a nanatural fermentation starter formulation was developed for manufacturing bread products by substituting baker's yeast with naturally fermented raisin extract and sourdough. Four experimental groups containing 2.5, 5.0, 7.5, 10% naturally fermented raisin extract per 2,000 g of flour were compared based on quality characteristics, including the fermentation power on dough expansion, specific volume, baking loss, water activity, color, textural characteristics, and internal surface appearance. The activities of the naturally fermented raisin extract were examined in terms of pH changes, total titratable acidity, brix, and viable yeast counts. The raisin extract, which was cultured for 7 days at 30$^{\circ}C$, smelled of alcohol and produced $CO_2$. Yeast were also found in the extract after separation. As the incubation time of the raisin extract and sourdough increased, pH decreased, while total titratable acidity increased. The brix of the raisin extract increased until the $2^{nd}$ day of fermentation, and viable yeast counts increased until the $5^{th}$ day however, these gradually decreased by the $7^{th}$ day. The fermenting power on dough expansion increased in the bread with increasing incubation time. The bread samples containing 7.5% and 10% raisin extract had significantly higher specific volumes than the other samples. Baking loss was minimal with the 2.5% extract substitution. In analyzing the crumb, water activity, redness, and yellowness were highest in the 10.0% raisin extract bread samples, and lightness was maximal in the 5.0% group. In terms of textural characteristics, hardness was lowest with the 2.5% extract substitution. Gumminess, springiness, and chewiness were not significantly different among the bread samples. Cohesiveness was highest at the 7.5% extract substitution level, and resilience was lowest at the 10% level. In conclusion, based on the results, a natural fermentation starter formulated with 2.5% naturally fermented raisin extract (1 part raisins and 1.5 parts water) and 70% sourdough (1 part rye flour and 1 part water) has high potential as a baker's yeast substitute for making naturally fermented bread.

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The Fission Yeast Gene Encoding Monothiol Glutaredoxin 5 Is Regulated by Nitrosative and Osmotic Stresses

  • Kim, Hong-Gyum;Park, Eun-Hee;Lim, Chang-Jin
    • Molecules and Cells
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    • v.20 no.1
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    • pp.43-50
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    • 2005
  • Glutaredoxin (Grx) is a small, heat-stable redox protein acting as a multi-functional glutathione (GSH)-dependent disulfide oxidoreductase. We have cloned the monothiol Grx5 gene from the genomic DNA of the fission yeast Schizosaccharomyces pombe. It has 1,904 bp, with one intron, and encodes a putative protein of 146 amino acids with a molecular mass of 16.5 kDa. Recombinant Grx5 produced functional Grx in S. pombe cells. NO-generating sodium nitroprusside (SNP, 1.0 and 2.0 mM) and potassium chloride (KCl, 0.2 and 0.5 M) increased the synthesis of ${\beta}$-galactosidase from a Grx5-lacZ fusion gene, and transcription of Grx5 was also enhanced by SNP and KCl. Synthesis of ${\beta}$-galactosidase from the Grx5-lacZ fusion was lower in Pap1-negative TP108-3C cells than in wild type KP1 cells, and when Pap1 was overproduced in KP1 cells, the level of ${\beta}$-galactosidase increased. We also found that Pap1 is involved in the induction of Grx5 by SNP and KCl. S. pombe Grx5 may play a crucial role in responses to nitrosative and osmotic stresses.

Immunoactivity of Ginsenosides Re and Rg1 that Enhances Resistance of Mice Against Experimental Disseminated Candidiasis

  • Han, Yong-Moon;Jin, Byung-Suk;Ko, Sung-Kwon;Lee, Jue-Hee
    • Natural Product Sciences
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    • v.10 no.3
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    • pp.134-139
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    • 2004
  • In this study, an immunoactivity of panaxtriol ginsenosides Re and Rg1 against infection due to Candida albicans was investigated. The ginsenosides were extracted from Red Ginseng with 85% ethanol and heat-treatment and were analyzed by HPLC on water-acetonitrile as a mobile phase. The HPLC analysis revealed that the extract contained ginsenosides Re and Rg1, which were eluted as a combined peak. By agar diffusion susceptibility, the mixture of Re and Rg1 had no growth-inhibitory activity on C. albicans yeast cells. However, in animal tests BALB/c mice given the mixture of Re and Rg1 intraperitoneally (Lp.) before intravenous (Lv.) infection with live C. albicans yeast cells had longer mean survival times (MST) than MST of control mice groups that received only buffer solution instead of Re and Rg1. In experiments 60% of the ginsenosides-treated mice survived the entire duration of the 50-day observation. The Re and Rg1 mixture induced production of nitric oxide when interacted with RAW 264.7 macrophage cell line. In addition, the mixture caused morphological change of the macrophages. These data indicate that immunostimulation by the Re and Rg1 may be responsible for the protection of mice against disseminated candidiasis.

Mapping of the Interaction Domain of DNA Topoisomerase $II{\alpha}$ and $II{\beta}$ with Extracellular Signal-Regulated Kinase 2

  • Park, Gye-Hwa;Bae, Young-Seuk
    • BMB Reports
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    • v.34 no.1
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    • pp.85-89
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    • 2001
  • Both topoisomerase $II{\alpha}$ and $II{\beta}$ east as phosphoproteins in the cells. Recently it was reported that DNA topoisomerase $II{\alpha}$ associates with and is phosphorylated by the extracellular signal-regulated kinase 2 (ERK2). Also, ERK2 stimulates the activity of topoisomerase II by a phosphorylation-independent manner [Shapiro et al., (1999) Mol. Cell. Biol. 19, 3551-3560]. In this study, a yeast two-hybrid system was used to investigate the binding site between topoisomerase $II{\alpha}$ or $II{\beta}$ and ERK2. The two-hybrid test clearly showed that topoisomerase $II{\beta}$ residues 1099-1263, and topoisomerase $II{\alpha}$ residues 1078-1182, mediate the interaction with ERK2, and that the leucine zipper motifs of topoisomerase $II{\alpha}$ and $II{\beta}$ are not required for its physical binding to ERK2. Our results suggest that topoisomerase $II{\beta}$ residues 1099-1263, and topoisomerase $II{\alpha}$ residues 1078-1182, may be common binding sites for activator proteins.

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Expression and Secretion of Foreign Proteins in Yeast Using the ADH1 Promoter and 97 K Killer Toxin Signal Sequence

  • Hong, Seok-Jong;Kang, Hyen-Sam
    • BMB Reports
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    • v.31 no.2
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    • pp.123-129
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    • 1998
  • Foreign proteins, $endo-{\beta}-1,4-glucanase$ of Bacillus subtilis, preS1+S2 region of hepatitis B virus large surface antigen, human ${\beta}_2-adrenergic$ receptor ($h{\beta}_{2}AR$), and bovine growth hormone (bGH) were expressed in Saccharomyces cerevisiae and secreted into the medium. These proteins were expressed using the alcohol dehydrogenase I (ADH1) promoter of Saccharomyces cerevisiae and secreted by signal sequence of the 97 K killer toxin gene of doublestranded linear DNA plasmid (pGKL1) of S. cerevisiae. All these proteins underwent severe modifications; in particular, N-glycosylation in the case of $endo-{\beta}-1,4-glucanase$, $h{\beta}_2AR$, and preS1+S2. Seventy four percent of the expressed $endo-{\beta}-1,4-glucanase$ was secreted into the culture medium. Highly modified proteins were detected in the culture medium and in the cell. Expressed $h{\beta}_2AR$, which has seven transmembrane domains, remained in the cell. The degrees of secretion and modification and the states of proteins in the culture medium and in the cell were quite different. These results indicated that the nature of the protein has a critical role in its secretion and modifications.

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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.

Production of High Viscous Hyaluronic Acid Complex from Klebsiella sp. L-10 (Klebsiella sp. L-10에 의한 고점성 히아루론산 복합체의 생산)

  • Lee, Hyang-Suk;Choe, Yeong-Jun;Lee, Jong-Su
    • The Journal of Natural Sciences
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    • v.7
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    • pp.59-65
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    • 1995
  • A bacterium which produce hyaluronic acid complex was isolated from soil, and identified as Klebsiella sp. L-10. The maximal hyaluronic acid complex production was obtained when the strain was cultured at $37^{\circ}C$ for 18hrs with shaking in the optimal medium containing 0.1% yeast ext., 3% tryptone, 5% glucose, 10mM $K_2HPO_4$and $KH_2PO_4$, respectively and initial pH6.5 and the final hyaluronic acid complex production under the above condition was 1,400mg per liter of cultures.

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Isolation of Yeasts utilizing Phthalic Compounds as a Sole Carbon Source (프탈산 화합물을 탄소원으로 이용하는 효모의 분리 및 동정)

  • Lee, Young-Nam;Yu, Jeong-Hi;Lee, Sang-Sun
    • The Korean Journal of Mycology
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    • v.19 no.1
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    • pp.41-46
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    • 1991
  • Three isolates of yeast utilizing phthalic compounds as a sole carbon source were obtained from the surface waters exposed to the industrial effluents near Cheong Ju city. On the basis of microscopic observations on morphology and various biochemical characterizations, the three isolates were identifed as a species of Rhodotorula. Candida or Torulopsis. A number of aroma­tic chemicals including phthalic compounds would support the growth of these yeasts as a sole carbon source, Thus, the yeast isolates would have potentials in reduction of environmental burden due to industrial wastes of aromatic hydrocarbons.

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