• Title/Summary/Keyword: yeast treatment

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Trehalose Protects Activity of Anti-Yeast Substance Produced by Rahnella aquatilis Strain AY2000 (Trehalose에 의한 Rahnella aquatilis AY2000 균주가 생산하는 항효모성 물질의 활성보호)

  • Kang, Min-Jung;Lee, Jong-Hwan;Lee, Bok-Kyu;Kim, Kwang-Hyeon
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.361-364
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    • 2009
  • Rahnella aquatilis strain AY2000 produces an anti-yeast substance (AYS), however activity of the AYS tends to be decreased by heat. To investigate whether trehalose can protect AYS activity against heat, comparative studies on the AYS with and without trehalose were performed. After heat treatment at high temperatures ($50^{\circ}C-70^{\circ}C$), the AYS with trehalose had higher activity than the AYS without trehalose had. In case of AYS with trehalose (0.3-0.9M), activity of the AYS could be determined at ranging from $7.8\;{\mu}g/mL$ to $31.3\;{\mu}g/mL$ on S. cerevisiae by MTT method. Consequently, activity of the AYS after heating was well maintained by trehalose.

Two Flavonoid-Based Compounds from Murraya paniculata as Novel Human Carbonic Anhydrase Isozyme II Inhibitors Detected by a Resazurin Yeast-Based Assay

  • Sangkaew, Anyaporn;Samritsakulchai, Nawara;Sanachai, Kamonpan;Rungrotmongkol, Thanyada;Chavasiri, Warinthorn;Yompakdee, Chulee
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.552-560
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    • 2020
  • Human carbonic anhydrase (CA) isozyme II has been used as protein target for disorder treatment including glaucoma. Current clinically used sulfonamide-based CA inhibitors can induce side effects, and so alternatives are required. This study aimed to investigate a natural CA inhibitor from Murraya paniculata. The previously developed yeast-based assay was used to screen 14 compounds isolated from M. paniculata and identified by NMR analysis for anti-human CA isozyme II (hCAII) activity. Cytotoxicity of the compounds was also tested using the same yeast-based assay but in a different cultivation condition. Two flavonoid candidate compounds, 5, 6, 7, 8, 3', 4', 5'-heptamethoxyflavone (4) and 3, 5, 7, 8, 3', 4', 5'-heptamethoxyflavone (9), showed potent inhibitory activity against hCAII with a minimal effective concentration of 10.8 and 21.5 μM, respectively, while they both exhibited no cytotoxic effect, even at the highest concentration tested (170 μM). The results from an in vitro esterase assay of the two candidates confirmed their hCAII inhibitory activity with IC50 values of 24.0 and 34.3 μM, respectively. To investigate the potential inhibition mechanism of compound 4, in silico molecular docking was performed using the FlexX and SwissDock software. This revealed that compound 4 coordinated with the Zn2+ ion in the hCAII active site through its methoxy oxygen at a distance of 1.60 Å (FlexX) or 2.29 Å (SwissDock). The interaction energy of compound 4 with hCAII was -13.36 kcal/mol. Thus, compound 4 is a potent novel flavonoid-based hCAII inhibitor and may be useful for further anti-CAII design and development.

Expression of Fungal Phytase on the Cell Surface of Saccharomyces cerevisiae

  • Mo, Ae-Young;Park, Seung-Moon;Kim, Yun-Sik;Yang, Moon-Sik;Kim, Dae-Hyuk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.576-581
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    • 2005
  • Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. We have engineered the cell surface of the yeast. Saccharomyces cerevisiae, by anchoring active fungal phytase on its cell wall, in order to apply it as a dietary supplement containing bioconversional functions in animal foods and a whole cell bio-catalyst for the treatment of waste. The phytase gene (phyA) of Aspergillus niger with a signal peptide of rice amylase 1A (Ramy1A) was fused with the gene encoding the C-terminal half (320 amino acid residues from the C-terminus) of yeast ${\alpha}-agglutinin$, a protein which is involved in mating and is covalently anchored to the cell wall. The resulting fusion construct was introduced into S. cerevisiae and expressed under the control of the constitutive glyceraldehydes-3-phosphate dehydrogenase (GPD) promoter. Phytase plate assay revealed that the surface-engineered cell exhibited a catalytically active opaque zone which was restricted to the margin of the colony. Additionally, the phytase activity was detected in the cell fraction, but was not detected in the culture medium when it was grown in liquid. These results indicate that the phytase was successfully anchored to the cell surface of yeast and was displayed as its active form. The amount of recombinant phytase on the surface of yeast cells was estimated to be 16,000 molecules per cell.

Analysis of Biological Effect of DBD-type Non-thermal Atmospheric Pressure Plasma on Saccharomyces Cerevisiae

  • Park, Gyung-Soon;Baik, Ku-Yeon;Kim, Jung-Gil;Kim, Yun-Jung;Lee, Kyung-Ae;Choi, Eun-Ha;Uhm, Hwan-Sup;Jung, Ran-Ju;Cho, Kwang-Sup
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.08a
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    • pp.337-337
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    • 2011
  • Application of plasma technology on microbial sterilization has been frequently studied. In spite of accumulating number of studies, many have been focused on bacteria. Reports on eukaryotic yeasts and filamentous fungi are limited. In addition, mechanism of plasma effect still needs to be clarified. In this study, we analyzed the effect of non-thermal atmospheric pressure plasma on the budding yeast, Saccharomyces cerevisiae using DBD-type device. When yeast cells were exposed to plasma (at 2 mm distance) and then cultured on YPD-agar plate, number of cells survived (shown as colony) were reduced proportionally to exposure time. More than 50% reduction in number of colonies were observed after twice exposure of 5min. each. Colonies much smaller than those of control (no plasma exposure) were appeared after twice exposure of 5 min. each. It seems that small colonies are resulted from delayed cell growth due to the damage caused by plasma treatment. Microscopic analysis demonstrates that yeast cells treated with plasma for 5 min. twice have more rough and shrinked shape compared to oval shape with smooth surface of control.

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Immunological Features of Macrophages Induced by Various Morphological Structures of Candida albicans

  • Han, Kyoung-Hee;Park, Su Jung;Choi, Sun Ju;Park, Joo Young;Lee, Kyoung-Ho
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.1031-1040
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    • 2013
  • Candida albicans is a dimorphic fungus that commensally colonizes human mucosal surfaces. The aim of this study was to assess the role of different C. albicans morphologies in inducing pattern recognition receptors (PRRs) and cytokines in macrophages. Macrophages may respond to pathogen-associated molecular patterns via TLR2 and TLR4 by expressing cytokines. The hyphal transition of C. albicans was induced by 20% serum (S), RPMI-1640 (R), or $39^{\circ}C$ culture (H). Macrophages were then challenged with either yeast (Y) or different hyphae cultures of C. albicans, followed by RT-PCR and FACS analysis of PRRs expression. In addition, macrophages were stimulated with either yeast or different hyphae cultures of C. albicans used by RT-PCR and Bio-Plex analysis of cytokines production. Macrophages expressed high levels of TLR4 and dectin-1 after stimulation with Y cells. In contrast, stimulation with H or R cells strongly increased the expression of TLR2 and dectin-2. Stimulation with Y cells significantly enhanced the expression of IL-$1{\beta}$ and weakly increased the expression of IL-6 and IL-12. Stimulation with hyphal cells (S, R, and H) strongly increased IL-10 expression, but weakly reduced IL-$1{\beta}$ expression. The phagocytosis activity and NO production of macrophages were decreased upon treatment with hyphal cells compared with yeast, and depended on the length of hyphae. In summary, the yeast and hyphae forms of C. albicans resulted in an induction of different PRRs, with accompanying differences in immune cell cytokine profiles.

A Modified Process for Producing High Quantities of Bio-Germanium in Yeast and a Study of Its Oral Toxicity

  • Park, So-Young;Joo, Seong-Soo;Won, Tae-Joon;Chung, Jin-Woong;Lee, Sung-Hee;Oh, Sun-Woo;Lee, Do-Ik;Hwang, Kwang-Woo
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.78-82
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    • 2007
  • Since germanium has been shown to be beneficial for the treatment of diseases such as cancer and rheumatic arthritis, we developed an adapted process of bio-germanium preparation using inorganic germanium. In the present study we determined the optimal conditions for culturing yeast Saccharomyces cerevisiae (KCTC-1199), and the best concentrations of inorganic germanium for the adaptation process. The resulting method was successful at producing high quantities of germanium yeasts. The following are the culture conditions that obtained the highest level of productivity: an inorganic germanium concentration of 3,000-5,000 ppm, a pH of 6.5, a temperature of $35^{\circ}C$, and 20 hr of incubation time. In addition to this high-yield quantity study, we observed the acute oral toxicity of mice treated with Geranti Bio-Ge $Yeast^{(R)}$. We found no changes in body weight, or in the mortality between the control groups and the bio-germanium yeast group. There were also no digestive problems such as diarrhea that occurred in either group.

Microbiological Studles of Red pepper paste Fermentation (II) (고추장의 발효미생물에 관한 연구 2)

  • 이택수;신보규;이석건;유주현
    • Korean Journal of Microbiology
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    • v.9 no.2
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    • pp.55-60
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    • 1971
  • Among 50 yeasts of red pepper paste isolated and identified in the previous report, 12 strains of yeast which possess osmophilic and non-film characteristics were examined their growth rates and alcohol fermentation ability in red pepper medium. 5 strains of yeast possessing powerful growth rates and excellent alcohol fermentation ability were selected as excellent yeasts in this experiment. And their cultural conditions, salt resistance and red pepper concentration upon the growth were examined in this report. The results obtained were as follows ; 1) Among 12 strains of yeast, Saccharomyces cerevisiae group II(strain $D_1$), Saccharomyces oviformis($D_2$), Saccharomyces steineri($M_3$), Saccharomyces rouxii($D_9$) and Saccharomyces mellis($G_8$) were selected as excellent yeasts. 2) Among 5 strains of yeast selected, the strain $D_1$ and $D_2$showed the best alcohol fermentation ability and the best fermentative flavour. 3) The optimum temperature for growth of the strain $D_1$ and $D_2$ was $25^{\circ}C$$M_3$, $D_9$ and $G_8$ $30^{\circ}C$$D_1$ and $D_2$ was $62^{\circ}C$$M_3$,$D_9$ and $G_8$ ws 60.deg.C with treatment for 10 minutes. 4) The optimum pH for growth of all strains was around 5.0. 5) The strain $D_1$ and $M_3$were grown exceedingly well on the emdia containing 3 percent of NaCl and $D_2$,$D_9$ and $G_8$ on the media containing 6 percent of NaCl. 6) In the case of 5 percent of red pepper liquor added 5 percent of glucose was more nutritional than in the case of 5 percent glucose only. However, the growth rates of yeasts were restrained by the increase of red pepper concentration.

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Discovery and Functional Study of a Novel Genomic Locus Homologous to Bα-Mating-Type Sublocus of Lentinula edodes

  • Lee, Yun Jin;Kim, Eunbi;Eom, Hyerang;Yang, Seong-Hyeok;Choi, Yeon Jae;Ro, Hyeon-Su
    • Mycobiology
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    • v.49 no.6
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    • pp.582-588
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    • 2021
  • The interaction of mating pheromone and pheromone receptor from the B mating-type locus is the first step in the activation of the mushroom mating signal transduction pathway. The B mating-type locus of Lentinula edodes is composed of Bα and Bβ subloci, each of which contains genes for mating pheromone and pheromone receptor. Allelic variations in both subloci generate multiple B mating-types through which L. edodes maintains genetic diversity. In addition to the B mating-type locus, our genomic sequence analysis revealed the presence of a novel chromosomal locus 43.3 kb away from the B mating-type locus, containing genes for a pair of mating pheromones (PHBN1 and PHBN2) and a pheromone receptor (RCBN). The new locus (Bα-N) was homologous to the Bα sublocus, but unlike the multiallelic Bα sublocus, it was highly conserved across the wild and cultivated strains. The interactions of RcbN with various mating pheromones from the B and Bα-N mating-type loci were investigated using yeast model that replaced endogenous yeast mating pheromone receptor STE2 with RCBN. The yeast mating signal transduction pathway was only activated in the presence of PHBN1 or PHBN2 in the RcbN producing yeast, indicating that RcbN interacts with self-pheromones (PHBN1 and PHBN2), not with pheromones from the B mating-type locus. The biological function of the Bα-N locus was suggested to control the expression of A mating-type genes, as evidenced by the increased expression of two A-genes HD1 and HD2 upon the treatment of synthetic PHBN1 and PHBN2 peptides to the monokaryotic strain of L. edodes.

Glutathione Content and the Activities of Glutathione-Synthesizing Enzymes in Fission Yeast are Modulated by Oxidative Stress

  • Lee, Yuk-Young;Kim, Su-Jung;Park, Eun-Hee;Lim, Chang-Jin
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.248-251
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    • 2003
  • Glutathione (GSH) is an important factor in determining tolerance against oxidative stress in living organisms. It is synthesized in two sequential reactions catalyzed by ${\gamma}$-glutamylcysteine synthetase (GCS) and glutathione synthetase (GS) in the presence of ATP. In this work, the effects of three different oxidative stresses were examined on GSH content and GSH-related enzyme activities in the fission yeast Schizosaccharomyces pombe. GSH content in S. pombe was significantly enhanced by treatment with hydrogen peroxide, ${\beta}$-naphthoflavone (BNF) and tert-butylhydroquinone (BHQ). Simultaneously, they greatly induced GCS and GS activity. However, they did not have any effects on glutathione reductase activity. These results suggest that GCS and GS activities in S. pombe are up-regulated by oxidative stress.

Grape Seed Extract Protects Mice against Disseminated Candidiasis

  • Han, Yong-Moon
    • Natural Product Sciences
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    • v.9 no.2
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    • pp.60-63
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    • 2003
  • Effect of grape seed extract (GSE) against Candida albicans was examined under in-vitro and in-vivo conditions. The GSE was extracted in ethanol. In-vitro results from an agar diffusion susceptibility assay showed the GSE inhibited C. albicans growth. This anticandidal effect was at dose-dependency. In experiments with animals, mice that received the GSE (0.5 mg per mice), intravenously (i.v.), before i.v.-infection wish viable C. albicans yeast cells survived longer than diluent (buffer)-received control mice. In contrast, when GSE was given to mice after the mice were infected with the yeast cells, these mice showed a similar survival rate as compared to control mice that received no treatment with the GSE. Taken together, these data indicate that GSE has prophylactic effect but not therapeutic effect against disseminated candidiasis.