• Title/Summary/Keyword: yeast cells

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Effect of soybean meal on the alcohol fermentation of sugar-alcohol-tolerant Saccharomyces cerevisiae (내당 내알콜성 Saccharomyces cerevisiae의 알콜 발효에 미치는 soybean meal의 영향)

  • Rho, Min-Jeong;Park, Keung-Ho;Paik, Un-Hwa;Yu, Ju-Hyun
    • Applied Biological Chemistry
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    • v.34 no.1
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    • pp.61-66
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    • 1991
  • In order to improve the productivity of ethanol by sugar-alcohol-tolerant Saccharomyces cerevisiae D1, the effect of addition of soybean meal on the alcohol fermentation was investigated. The addition of soybean meal led tn the increase of the ethanol productivity and viable cell concentration. Increasing the mont of soybean meal increased the number of viable cells and the consumption percentage of glucose. The water-soluble fraction of soybean meal was nearly as effective as whole-soybean meal, whereas the lipidic fraction had no positive effect. The addition of 4% soybean meal increased the rate of ethanol production regardless of the initial concentrations of glucose. The rate of glucose consumption fermenting a soybean meal supplemented medium was higher than possible in a non-supplemented medium, either in the absence or in the presence of ethanol. But the percentage of ethanol inhibition of the glucose consumption rate was identical for supplemented md unsupplemented media. The increase of final ethanol concentration could not be attributed In an increase of ethanol tolerance of yeast cells but to the satisfaction of nutritional deficiencies.

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PRIP, a Novel Ins(1,4,5)P3 Binding Protein, Functional Significance in Ca2+ Signaling and Extension to Neuroscience and Beyond

  • Kanematsu, Takashi;Takeuchi, Hiroshi;Terunuma, Miho;Hirata, Masato
    • Molecules and Cells
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    • v.20 no.3
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    • pp.305-314
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    • 2005
  • Investigation of chemically synthesized inositol 1,4,5-trisphosphate [$Ins(1,4,5)P_3$] analogs has led to the isolation of a novel binding protein with a molecular size of 130 kDa, characterized as a molecule with similar domain organization to phospholipase C-${\delta}1$ (PLC-${\delta}1$) but lacking the enzymatic activity. An isoform of the molecule was subsequently identified, and these molecules have been named PRIP (PLC-related, but catalytically inactive protein), with the two isoforms named PRIP-1 and -2. Regarding its ability to bind $Ins(1,4,5)P_3$ via the pleckstrin homology domain, the involvement of PRIP-1 in $Ins(1,4,5)P_3$-mediated $Ca^{2+}$ signaling was examined using COS-1 cells overexpressing PRIP-1 and cultured neurons prepared from PRIP-1 knock-out mice. Yeast two hybrid screening of a brain cDNA library using a unique N-terminus as bait identified GABARAP ($GABA_A$ receptor associated protein) and PP1 (protein phosphatase 1), which led us to examine the possible involvement of PRIP in $GABA_A$ receptor signaling. For this purpose PRIP knock-out mice were analyzed for $GABA_A$ receptor function in relation to the action of benzodiazepines from the electrophysiological and behavioral aspects. During the course of these experiments we found that PRIP also binds to the b-subunit of $GABA_A$ receptors and PP2A (protein phosphtase 2A). Here, we summarize how PRIP is involved in $Ins(1,4,5)P_3$-mediated $Ca^{2+}$ signaling and $GABA_A$ receptor signaling based on the characteristics of binding molecules.

Microarray analysis of gene expression in raw cells treated with scolopendrae corpus herbal-acupuncture solution (蜈蚣(오공) 약침액(藥鍼液)이 LPS로 처리된 RAW 세포주(細胞柱)의 유전자(遺傳子) 발현(發顯)에 미치는 영향(影響))

  • Bae, Eun-Hee;Lee, Kyung-Min;Lee, Bong-Hyo;Lim, Seong-Chul;Jung, Tae-Young;Seo, Jung-Chul
    • Korean Journal of Acupuncture
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    • v.23 no.3
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    • pp.133-160
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    • 2006
  • Objectives : Scolopendrae Corpus has a broad array of clinical applications in Korean medicine, including treatment of inflammatory conditions such as arthritis. To explore the global gene expression profiles in human Raw cell lines treated with Scolopendrae Corpus herbal-acupuncture solution (SCHAS), cDNA microarray analysis was performed. Methods : The Raw 264.7 cells were treated with lipopolysaccharide (LPS), SCHAS, or both. The primary data was normalized by the total spots of intensity between two groups, and then normalized by the intensity ratio of reference genes such as housekeeping genes in both groups. The expression ratio was converted to log2 ratio. Normalized spot intensities were calculated into gene expression ratios between the control and treatment groups. Greater than 2 fold changes between two groups were considered to be of significance. Results : Of the 8 K genes profiled in this study, with a cut-off level of two-fold change in the expression, 20 genes (BCL2-related protein A1, MARCKS-like 1, etc.) were upregulated and 5 genes (activated RNA polymerase II transcription cofactor 4, calcium binding atopy-related autoantigen 1, etc.) downregulated following LPS treatment. 139 genes (kell blood group precursor (McLeod phenotype), ribosomal protein S7, etc.) were upregulated and 42 genes (anterior gradient 2 homolog (xenopus laevis), phosphodiesterase 8B, etc.) were downregulated following SCHAS treatment. And 10 genes (yeast saccharomyces cerevisiae intergeneic sequence 4-1, mitogen-activated protein kinase 1, etc.) were upregulated and 8 genes (spermatid perinuclear RNA binding protein, nuclear receptor binding protein 2, etc.) were downregulated following co-stimulation of SCHAS and LPS. Discussions : It is thought that microarrays will play an ever-growing role in the advance of our understanding of the pharmacological actions of SCHAS in the treatment of arthritis. But further studies are required to concretely prove the effectiveness of SCHAS.

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A putative prolyl tRNA synthetase is involved in pheromone induction in Schizosaccharomyces pombe (Schizosaccharomyces pombe의 pheromone 유도와 연관된 prolyl tRNA synthetase)

  • Kim, Daemyung
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.309-319
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    • 2018
  • Previously, six Schizosaccharomyce pombe mutants that induce pheromone even in the presence of nitrogen source were isolated from a bank of temperature sensitive mutants. In this report, one of these mutants, pws6 was further characterized. The pheromone induction in pws6 mutant cells was specific to nutrient: the M-factor pheromone was induced without nitrogen starvation but not without glucose starvation. This result suggests that the pws6 mutant might have a specific defect in the pathway for nitrogen starvation. The pws6 mutant induces P-factor pheromone as well as M-factor without starvation of nitrogen in temperature sensitive mode, suggesting that the pheromone induction phenotype of pws6 mutation is not cell-type specific. From cloning of the $pws6^+$ gene by complementation of the temperature sensitive growth defect, three plasmids containing 8.1 kb, 3.3 kb, and 4.8 kb yeast DNA were recovered. These plasmids complement the growth defect of the pws6 mutant by 100%, 70%, and 10~20%, respectively. The abilities of these plasmids to complement pheromone induction phenotype of pws6 mutant cells were correlated well with the efficiencies of complementation of the growth defect. With comparison of their open reading frames to the complementation efficiencies, it is concluded that the open reading frame, SPBC19C7.06 is responsible for the complementation of temperature sensitive phenotype of the pws6 mutant. This open reading frame, named prs1, contains one long exon with no intron and encodes a putative prolyl tRNA synthetase. The putative Prs1 protein exhibits significant similarities to the prolyl tRNA synthetases of other species.

Effect of High Hydrostatic Pressure Treatment on Extension of Shelf Life of Draft Makgeolli (초고압처리가 생막걸리의 저장기간 연장에 미치는 영향)

  • Choi, H.S.;Kang, C.S.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.22 no.2
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    • pp.5-15
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    • 2020
  • To extend the shelf life of draft makgeolli, UV irradiation and high hydrostatic pressure were conducted. UV irradiation did not reduce the total cell number in makgeolli, but high hydrostatic pressure treated drastically decreased. The number of viable cells in makgeolli was 2.2-5.8 × 107 CFU/mL, but when treated with 300 MPa pressure for 1 minute, it decreased to 1.4-10 × 103 CFU/mL, with 400 MPa to 4-68 CFU/mL, and with 500 MPa to under 40 CFU/mL. Yeast died at 400 MPa pressure, but Bacillus amyloliquefaciens and Rummeliibacillus stabekisii survived at 500 MPa pressure. The makgeolli treated with 400 and 500 MPa pressure did not increase the number of cells even when stored at room temperature (25℃) for 30 days, and changes in alcohol, acidity, and amino acid vlaue were also suppressed. However, when stored for more than 30 days at room temperature, the acidity and amino acid levels of makgeolli increased, making it difficult to drink. At low temperature (4℃) storage, the quality change of makgeolli was suppressed until 70 days of storage, so it had a taste similar to that of fresh makgeolli.

Medium optimization for growth of Bacillus amyloliquefaciens ISP-5 strain and evaluation of plant growth promotion using lettuce (Bacillus amyloliquefaciens ISP-5 균주의 배지 최적화 및 상추를 이용한 식물 생장 촉진 평가)

  • Kang-Hyun Choi;Sun Il Seo;Haeseong Park;Ji-hwan Lim;Pyoung Il Kim
    • Journal of Plant Biotechnology
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    • v.49 no.4
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    • pp.356-361
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    • 2022
  • Bacillus sp. is a useful strain for agriculture because it promotes plant growth and controls plant pathogens through a variety of mechanisms. In this study, we obtained a microbial preparation with a high number of viable cells by culturing newly isolated soil bacteria on an optimized medium. Subsequently, we applied this preparation to lettuce to enhance its growth and yield. First, B. amyloliquefaciens ISP-5 was isolated from soil. Next, optimization of culture medium was carried out using 5 L scale fermenters. When culturing B. amyloliquefaciens ISP-5 on this optimized medium, the number of viable cells was approximately 1000 times higher than that obtained from culturing on the commercial medium. Afterwards, the plant growth promotion properties of the ISP-5 strain were evaluated using lettuce as a test plant. Foliar spray treatment of lettuce was carried out by inoculating half the standard concentration suspension (0.5 × 107 cfu/ml). As a result, leaf width increased by 8.6% and leaf length increased by 12.9% compared to the control group. Live weight also increased by 24.2% and dry weight by 23.9%. Considering the results from field test, B. amyloliquefaciens ISP-5 showed potential as a plant growth-promoting bacteria.

Isolation and Characterization of mas1+ of Schizosaccharomyces pombe, a Homologue of Human CIP29/Hcc-1 Involved in the Regulation of Cell Division (세포분열에 관여하는 인간의 CIP29/Hcc1 유전자와 상동성을 가지는 분열형 효모의 새로운 유전자 mas1+의 특성분석)

  • Cha, Jae-Young;Shin, Sang-Min;Ha, Se-Eun;Lee, Jung-Sup;Park, Jong-Kun
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1666-1677
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    • 2011
  • The regulation of gene expression plays an important role in cell cycle controls. In this study, a novel gene, the $mas1^+$($\underline{mi}$tosis $\underline{as}$sociated protein) gene, a homolog of human CIP29/Hcc1, was isolated and characterized from fission yeast Schizosaccharomyces pombe (S. pombe) using a gene-specific polymerase chain reaction. The isolated gene contained a complete open reading frame capable of encoding 245 amino acid residues with a typical promoter, as judged by nucleotide sequence analysis. It was also found that a PCB ($\underline{p}$ombe cell $\underline{c}$ycle $\underline{b}$ox) is located in the promoter region, which controls M-$G_1$ specific transcription in S. pombe. The quantitative analysis of the $mas1^+$ transcript against $adh1^+$ showed that the pattern of expression is similar to that of the septation index. Cytokinesis of mas1 mutant was greatly delayed at $25^{\circ}C$ and $36^{\circ}C$, and a large number of multi-septate cells were produced. The mas1 mutant had 2C, 4C and 6C DNA contents, as determined by FACS analysis. In addition, the number of multi-septate cells significantly increased. When cells were cultured in nitrogen starvation medium to increase proliferation, the abnormal phenotypes of mas1 mutant dramatically increased. These phenotypes could be rescued by an overexpression of the $mas1^+$ gene. The mas1 protein localized in the nuclei of S. pombe and human HeLa cells, as evidenced by Mas1-EGFP signals. The abnormal growth pattern and the morphology of mas1 mutant were complemented by a plasmid carrying human CIP29/Hcc-1cDNA. In addition, CIP29 /Hcc-1 transcript level increased in active cell proliferation stages in the developing mouse embryos. These results indicate that the $mas1^+$ ishomologous to the human CIP29/Hcc1 gene and is involved in cytokinesis and cell shape control.

PDZ Domain-containing Proteins at Autotypic Junctions in Myelinating Schwann Cells (수초화 슈반세포 autotypic 세포연접의 PDZ 도메인 보유 단백질)

  • Han, Seongjohn;Park, Hyeongbin;Hong, Soomin;Lee, Donghyun;Choi, Maro;Cho, Jeongmok;Urm, Sang-Hwa;Jang, Won Hee;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.25 no.1
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    • pp.101-112
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    • 2015
  • A type of cell junction that is formed between different parts within the same cell is called autotypic cell junction. Autotypic junction proteins form tight junctions found between membrane lamellae of a cell, especially in myelinating glial cells. Some of them have postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domains, which interact with the carboxyl (C)-terminal PDZ-binding motif of other proteins. PDZ domains are protein-protein interaction modules that play a role in protein complex assembly. The PDZ domain, which is widespread in bacteria, plants, yeast, metazoans, and Drosophila, allows the assembly of large multi-protein complexes. The multi-protein complexes act in intracellular signal transduction, protein targeting, and membrane polarization. The identified PDZ domain-containing proteins located at autotypic junctions include zonula occludens-1 (ZO-1), ZO-2, pals-1-associated tight junction protein (PATJ), multi-PDZ domain proteins (MUPPs), membrane-associated guanylate kinase inverted 2 (MAGI2), and protease-activated receptor (PAR)-3. PAR-3 interacts with atypical protein kinase C and PAR-6, forming a ternary complex, which plays an important role in the regulation of cell polarity. MAGI2 interacts with ${\alpha}$-amino-3-hydroxyl-5-methyl-4-isoxazole propionate (AMPA) receptor at excitatory synapses. PATJ is detected in paranodal loops associated with claudin-1. On the other hand, MUPP1 is found in mesaxons and Schmidt-Lanterman incisures with claudin-5. ZO-1, ZO-2, and PAR-3 are found at all three sites. Different distributions of PDZ domain-containing proteins affect the development of autotypic junctions. In this review, we will describe PDZ domain-containing proteins at autotypic tight junctions in myelinating Schwann cells and their roles.

Characterizations of Kefir Grains in Fermented Whey and Their Effects on Inflammatory Cytokine Modulation in Human Mast Cell-1 (HMC-1) (Kefir grain에 의한 유청발효액의 특성과 human mast cell-1 (HMC-1)에서 염증 cytokine 조절에 미치는 영향)

  • Son, Ji Yoon;Park, Young W.;Renchinkhand, Gereltuya;Han, Jung Pil;Bum, Jin Woo;Paik, Seung-Hee;Lee, Jo Yoon;Nam, Myoung Soo
    • Journal of Life Science
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    • v.26 no.6
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    • pp.689-697
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    • 2016
  • Kefir is an acidic-alcoholic fermented milk product originating from the Caucasian mountains. Kefir has long been known for its probiotic health benefits, including its immunomodulatory effects. The objectives of this study were to investigate the properties of a fermented whey product and to examine the effects of kefir grains on the in vitro immune-modulation of human mast cell-1 (HMC-1). The results showed that the whey fermented by kefir grains contained the maximum lactic acid bacteria and yeast for 16 hr by 1.83×108 and 6.5×105 CFU/ml, respectively, and lactose and whey proteins were partially hydrolyzed. The experimental whey fermented by kefir grains exhibited an in vitro anti-inflammatory effect on the HMC-1 line for 8, 16, and 24 hr, and this effect induced the expression of interleukin (IL)-4 as a pro-inflammatory cytokine, but not for 48 hr by RT-PCR in HMC-1 cells. In addition, the same phenomenon was observed for the expression of IL-8 as a pro-inflammatory cytokine by the kefir-fermented whey during the same periods of 8-48 hr under the same conditions. These cytokines resulted in the production of IL-4 at 20-25 ng in HMC-1 cells for 8, 16, and 24 hr, whereas 5 ng was produced for 48 hr by the fermented whey. In contrast, IL-8 was produced at 15-20 ng in HMC-1 cells during 4, 8, 16, and 24 hr, while 7 ng was produced at 48 hr. It was concluded that the whey fermented by kefir grains possesses a potential anti-inflammatory function, which could be used for an industrial application as an ingredient of functional foods and pharmaceutical products.

Anti-inflammatory Activity of Antimicrobial Peptide Protaetiamycine 2 Derived from the Protaetia brevitarsis seulensis (흰점박이꽃무지 유래 항균 펩타이드 프로테티아마이신 2의 항염증활성)

  • Lee, Joon Ha;Baek, Minhee;Lee, Hwa Jeong;Kim, In-Woo;Kim, Sun Young;Seo, Minchul;Kim, Mi-Ae;Kim, Seong Hyun;Hwang, Jae Sam
    • Journal of Life Science
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    • v.29 no.11
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    • pp.1218-1226
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    • 2019
  • The white-spotted flower chafer Protaetia brevitarsis seulensis is a medicinally beneficial and important edible insect species. We previously performed an in silico analysis of the Protaetia brevitarsis seulensis transcriptome to identify putative antimicrobial peptides and then tested their antimicrobial and hemolytic activities. These peptides had potent antimicrobial activities against bacteria and yeast without inducing hemolysis. In the present study, the cationic antimicrobial peptide, protaetiamycine 2, was selected for further assessment of its anti-inflammatory properties in mouse macrophage Raw264.7 cells. Protaetiamycine 2 treatment of Raw264.7 cells suppressed LPS-induced nitric oxide production and reduced the expression of inducible nitric oxide synthase and cyclooxygenase-2, as determined by real-time PCR and western blotting. The expression of proinflammatory cytokines ($TNF-{\alpha}$, IL-6, and $IL-1{\beta}$) was also attenuated through the MAPKs and $NF-{\kappa}B$ signaling. We also confirmed that protaetiamycine 2 bound to bacterial cell membranes by a specific interaction with LPS. Collectively, these data obtained from LPS-induced Raw264.7 cells indicated that protaetiamycine 2 could have both antimicrobial and anti-inflammatory properties.