• Title/Summary/Keyword: yeast cells

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Improved Production of Live Cells of Lactobacillus rhamnosus by Continuous Cultivation using Glucose-yeast Extract Medium

  • Ling Liew Siew;Mohamad Rosfarizan;Rahim Raha Abdul;Wan Ho Yin;Ariff Arbakariya Bin
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.439-446
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    • 2006
  • In this study, the growth kinetics of Lactobacillus rhamnosus and lactic acid production in continuous culture were assessed at a range of dilution rates $(0.05 h^{-1}\;to\;0.40h^{-1})$ using a 2L stirred tank fermenter with a working volume of 600ml. Unstructured models, predicated on the Monod and Luedeking-Piret equations, were employed to simulate the growth of the bacterium, glucose consumption, and lactic acid production at different dilution rates in continuous cultures. The maximum specific growth rate of L. rhamnosus, ${\mu}_{max}$, was estimated at $0.40h^{-1}$I, and the Monod cell growth saturation constant, Ks, at approximately 0.25g/L. Maximum cell viability $(1.3{\times}10^{10}CFU/ml)$ was achieved in the dilution rate range of $D=0.28h^{-1}\;to\;0.35h^{-1}$. Both maximum viable cell yield and productivity were achieved at $D=0.35h^{-1}$. The continuous cultivation of L. rhamnosus at $D=0.35h^{-1}$ resulted in substantial improvements in cell productivity, of 267% (viable cell count) that achieved via batch cultivation.

Application of Saccharomyces rouxii for the Production of Non-alcoholic Beer

  • Sohrabvandi, Sarah;Razavi, Seyed Hadi;Mousavi, Seyed Mohammad;Mortazavian, Amir;Rezaei, Karamathollah
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1132-1137
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    • 2009
  • Successive application of Saccharomyces cerevisiae DSM 70424 and Saccharomyces rouxii DSM 2535 or DSM 2531 in the production of non-alcoholic beer was investigated. The aim of the study was to consider the impact of the 2 mentioned strains of S. rouxii on the reduction of alcohol content in wort fermented at 12 or $24^{\circ}C$ for 96 hr, applying periodic aeration. The 2 S. rouxii strains were added at the $48^{th}$ hr of fermentation after thermal inactivation of S. cerevisiae cells. The greatest alcohol decrease rate was observed for the treatment containing S. rouxii DSM 2535-fermented at $24^{\circ}C$ (from 1.56 to 0.36%). The concentration of acetaldehyde, diacetyl, and 2,3-pentandione, that have a key role in appearance of 'wort' and 'buttery' off flavors, were significantly lower in S. rouxii-containing treatments fermented at $24^{\circ}C$. S. rouxii-containing treatment fermented at $24^{\circ}C$ showed slightly lower overall flavor acceptability compared to S. cerevisiae-containing treatment fermented at the same temperature. Such score was improved for the products obtained at $12^{\circ}C$.

Immunostimulating Activity of Beta-Glucan Isolated from the Cell Wall of Mutant Saccharomyces cerevisiae, and Its Anti-Tumor Application in Combination with Cisplatin (Saccharomyces cerevisiae 변이주 세포벽 유래의 베타글루칸의 면역활성 및 Cisplatin과의 병용에 의한 항암 상승작용)

  • Kim, Wan-Jae;Yoon, Taek-Joon;Kim, Dong-Woo;Moon, Won-Kook;Lee, Kwang-Ho
    • The Korean Journal of Food And Nutrition
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    • v.23 no.2
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    • pp.141-146
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    • 2010
  • Cisplatin(cis-diamminedichloroplatium) is one of the most effective anti-cancer drugs being clinically used in the treatment of solid tumors. Despite its therapeutic benefits, its use in clinical practice is often limited because of dose related toxicity. It is known that yeast cell wall beta-glucans possess immuno-modulating properties, which allows for their application in antitumor therapy. IS2 is a kind of beta-glucan derived from the cell wall of mutated Saccharomyces cerevisiae, which exhibits anti-cancer activity in vitro and in vivo. The present study explored the possibility of combination therapy of IS2 with cisplatin. In experimental metastasis of colon26-M3.1 cells, prophylactic intravenous administration of IS-2 in combination with cisplatin effectively inhibited tumor metastasis compared with cisplatin alone or IS-2 treatment in vivo. IS-2 effectively enhanced Th1 type cytokines including IFN-$\gamma$, IL-2, IL-12 and GM-CSF. Simultaneously, this combined treatment inhibited production of Th2 type cytokines compared with control. These results suggested that IS-2 can be applied in combination therapy with anti-cancer drugs to minimize their side effects.

Enhanced Production, Purification, and Partial Characterization of Lacticin BH5, a Kimchi Bacteriocin Produced by Lactococcus lactis BH5

  • Paik, Hyun-Dong;Hyun, Hyung-Hwan;Pyun, Yu-Ryang;Ahn, Cheol;Hur, Ji-Woon;Kim, Tae-Seok;Yeo, Ick-Hyun
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.53-60
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    • 2000
  • Strain BH5 was isolated from naturally fermented Kimchi and identified as a bacteriocin producer, which has bactericidal activity against Micrococcus flavus ATCC 10240. Strain BH5 was identified tentatively as Lactococcus lactis by the API test and some characteristics. Lactococcus lactis BH5 showed a broad spectrum of activity against most of the non-pathogenic and pathogenic microorganisms tested by the modified deferred method. The activity of lacticin BH5, named tentatively as the bacteriocin produced by Lactococcus lactis BH5, was detected at the mid-log growth phase, reached its maximum during the early stationary phase, and decreased after the late stationary phase. Lacticin BH5 also showed a relatively broad spectrum of activity against non-pathogenic and pathogenic microorganisms as tested by the spot-on-lawn method. Its antimicrobial activity on sensitive indicator cells was completely disappeared by protease XIV or ${\alpha}$-chymotrypsin. The inhibitory activities of lacticin BH5 were detected during treatments up to 100$^{\circ}C$ for 30 min. Lacticin BH5 was very stable over a pH range of 2.0 to 9.0 and was stable with all the organic solvents examined. The cell concentration and bacteriocin production in strain BH5 were maximum when grown at 30$^{\circ}C$ in a modified MRS medium supplemented with 0.5% tryptone, 1.0% yeast extract, and 0.5% beef extract as nitrogen sources. It demonstrated a typical bactericidal mode of inhibition against Micrococcus flavus ATCC 10240. Lacticin BH5 was purified through ammonium sulfate precipitation, ethanol precipitation, and CM-Sepharose column chromatography. The apparent molecular mass of lacticin BH5 was estimated to be in the region of 3.7 kDa, by the direct detection of bactericidal activity after SDS-PAGE. Mutant strain NO141 which was isolated by nitrosoguanidine mutagenesis produced about 4 fold more bacteriocin than the wild type.

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PtdIns(3,5)P2 5-phosphatase Fig4 Interacts with Kinesin Superfamily 5A (KIF5A) (PI(3,5)P2 5-phosphatase Fig4와 Kinesin superfamily 5A (KIF5A)의 결합)

  • Jang, Won Hee;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.24 no.1
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    • pp.14-19
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    • 2014
  • Kinesin-1 consists of two heavy chains (KHCs), also called KIF5s, and two light chains (KLCs) that form a heterotetrameric complex. Here, we demonstrate the binding of a neuronal KHC, KIF5A, to the carboxyl (C)-terminal tail region of Fig4 (also known as Sac3), a phosphatase that removes the 5-phosphate from phosphatidylinositol-3,5-bisphosphate ($PtdIns(3,5)P_2$). Fig4 bound to the C-terminal region of KIF5A but not to other KHCs (KIF5B and KIF5C) and KLC1 in yeast two-hybrid assays. The interaction was further confirmed in a glutathione S-transferase pull-down assay and by co-immunoprecipitation. Anti-KIF5A antibody co-immunoprecipitated Fig4 with KIF5A from mouse brain extracts. These results suggest that kinesin-1 could transport the Fig4-associated protein complex or cargo in cells.

Study on the Potential Hazard Analysis by Physicochemical Quality and Microbiological Safety Comparison of Commercial Sauces in South Korea (시판 소스의 이화학적, 미생물적 품질 특성 조사를 통한 잠재적 위해도 연구)

  • Kang, Hyun Seok;Kim, Hyun Soo;Ahn, Joung Jwa;Yun, Tae Mi;Hwang, Tae Young
    • Journal of the East Asian Society of Dietary Life
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    • v.24 no.6
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    • pp.845-852
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    • 2014
  • The quality of commercial sauce products was evaluated through pH, soluble solid content, salinity, water activity and microbial analyses. The pH of sauces was 2.38~5.30, soluble solids were between 6.03 and 71.67, and distributions of salt were 0.23~5.00% in 32 commercial sauce products. In addition, water activity of vinegar red pepper sauce and spicy soft tofu stew stock were determined 0.773 and 0.988, respectively. Yeast, mold, Staphylococcus aureus, E. coli and coliform were not detected in any sauces. Higher level of total viable cells (TVC) resulted in pH over 4.2. TVC of shelf-stable sauces was 1.0~3.6 log CFU/g. TVC of seven sauce products was classified as non-potentially hazardous foods by temperature controlled for safety standard (TCS), even though levels were over acceptable guidelines of the USDA (3 log CFU/g). These results indicate that the standard and classification of commercial sauce products should be modified and controlled strictly.

Production of Aminoglycoside-3'-Phosphotransferase by the Fed-Batch Cultivation of Mutant Obtained from E. coli ATCC 21990 (E.coli ATCC 21990 변이주의 유가배양법에 의한 Aminoglycoside-3'-Phosphotransferase 생산)

  • 김기태;김학주;김계원;나규흠;양중익;김수일
    • Microbiology and Biotechnology Letters
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    • v.19 no.5
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    • pp.491-496
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    • 1991
  • To maximize the production of aminoglycoside-3'-phosphotransferase of E. coli ATCC 21990 carrying R factor which encodes aminoglycoside-3'-phosphotransferase (APH(3')) phosphorylating the 3'-hydroxyl group of aminoglycoside, mutants M1 and M2, media composition and several factors affecting the enzyme production during fermentation were studied. Although the specific activity of APH(3') produced by a mutant M1 was increased as much as four times than that of E. coii ATCC 21990, the growth rate was decreased. The increase of the enzyme production was obtained by increased biomass during fermentation. A mutant M2 was obtained to increase the cell growth rate. Mutant M2 cells were cultivated with optimal media and pure oxygen gas in a fed-batch mode of fermentor operation. The specific activity of APH(3') was decreased, but total enzyme activity of APH(3') was increased as much as two point five times than that of mutant MI.

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Physiological Effects of GLT1 Modulation in Saccharomyces cerevisiae Strains Growing on Different Nitrogen Sources

  • Brambilla, Marco;Manuela Adamo, Giusy;Frascotti, Gianni;Porro, Danilo;Branduardi, Paola
    • Journal of Microbiology and Biotechnology
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    • v.26 no.2
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    • pp.326-336
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    • 2016
  • Saccharomyces cerevisiae is one of the most employed cell factories for the production of bioproducts. Although monomeric hexose sugars constitute the preferential carbon source, this yeast can grow on a wide variety of nitrogen sources that are catabolized through central nitrogen metabolism (CNM). To evaluate the effects of internal perturbations on nitrogen utilization, we characterized strains deleted or overexpressed in GLT1, encoding for one of the key enzymes of the CNM node, the glutamate synthase. These strains, together with the parental strain as control, have been cultivated in minimal medium formulated with ammonium sulfate, glutamate, or glutamine as nitrogen source. Growth kinetics, together with the determination of protein content, viability, and reactive oxygen species (ROS) accumulation at the single cell level, revealed that GLT1 modulations do not significantly influence the cellular physiology, whereas the nitrogen source does. As important exceptions, GLT1 deletion negatively affected the scavenging activity of glutamate against ROS accumulation, when cells were treated with H2O2, whereas Glt1p overproduction led to lower viability in glutamine medium. Overall, this confirms the robustness of the CNM node against internal perturbations, but, at the same time, highlights its plasticity in respect to the environment. Considering that side-stream protein-rich waste materials are emerging as substrates to be used in an integrated biorefinery, these results underline the importance of preliminarily evaluating the best nitrogen source not only for media formulation, but also for the overall economics of the process.

Changes of Microbiological and General Quality Characteristics of Gamma Irradiated Kochujang(Fermented Hot Pepper Paste) (감마선 조사 고추장의 미생물 및 일반품질 특성 변화)

  • Kim, Dong-Ho;Yook, Hong-Sun;Youn, Kyu-Chun;Sohn, Cheon-Bae;Byun, Myung-Woo
    • Korean Journal of Food Science and Technology
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    • v.33 no.1
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    • pp.72-77
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    • 2001
  • The effect of gamma-irradiation on the microbiological and general quality changes of the kochujang(Korean hot pepper paste) was studied. Kochujang was prepared, irradiated at 5, 10 and 20 kGy, and then stored at $25^{\circ}C$ for twelve weeks. The results showed that the Bacillus were decreased by 5 log cycles with dose of 20 kGy with 3.94 kGy of $D_{10}$ value. Yeast and Lactobacillus cells were nearly eliminated by 10 kGy. The formation of $NH_2-nitrogen$ and reducing sugar, enzyme activity, acid production and browning were not affected by gamma irradiation. Therefore, it was considered that the kochujang treated with gamma irradiation maintained better microbial quality than that of the control with storage.

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Characterization of a novel protein interacting with rat large-conducatance $Ca^{2+}$-actived $K^+$ channel $a\lpha$-subunit rSlo

  • Aegyoung Cho;Lee, Kwang-Hee;Sungmin Song;Bongwoon Hwang;Jung, Yong-Keun;Park, Chul-Seung
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.43-43
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    • 2003
  • Large-conductance $Ca^{2+}$-actived $K^{+}$ channels ($BK_{Ca}$ channels) play a key role in setting the pace of contractile activity in muscle and are involved in the regulation of neurotransmitter release in neuron. $BK_{Ca}$ channels are activated by depolarizing membrane potential and the elevated level of intracellular calcium. Using yeast-two hybrid assay, we have identified a novel protein interacting with the cytosolic carboxyl terminus of rSlo, the brain isoform of rat large-conductance $Ca^{2+}$-activated $K^{+}$ channel $\alpha$-subunit. The novel gene encodes 51 kDa protein and is named as SIRK(rSlo-interacting RGS-like protein). SIRK is expressed in various tissues and localized in the cytosolic and the membrane fraction. Biochemical and immunological studies indicated that SIRK physically interacted with the cytosolic region of rSlo. To investigate whether SIRK can modulate the activity of rSlo, GFP-fused SIRK and rSlo were transiently transfected into COS-7 cells and the effects of SIRK was studied using electrophysiological means. We concluded that the overexpression of SIRK alters the surface expression of rSlo channel with only a limited effect on the biophysical characteristics of the channel.the channel.

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