• Title/Summary/Keyword: culture supernatants

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고삼투압이 재조합 Erythropoietin의 생산과 당쇄구조에 미치는 효과

  • Jeong, Yeon-Tae;Kim, Jeong-Hoe
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.221-224
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    • 2001
  • Effect of hyperosmotic pressure on growth of recombinant Chinese hamster 。 vary cells and Erythropoietin (EPO) production was investigated. Cells were cultivated in batch modes at various osmolalities. When the osmolality increased from 314 to 463mOsm/Kg, specific EPO productivity (qp) was increased up to 1.6-fold but cell growth was inhibited. EPO has a complex oligosaccharide structure that plays an important role in biological activity in vivo. To investigate the influence of hypoerosmotic pressure on the glycosylation, structural analysis of oligosaccharide was calTied out. Recombinant human EPO was produced by CHO cells grown under various osmotic pressure and purified from culture supernatants by heparin-sepharose affinity column and immunoaffinity column. N-linked oligosaccharides were released enzymatically and isolated by paper chromatography. The isolated oligosaccharides were labeled with fluorescent dye, 2-aminobenzamide and analyzed with MonoQ anion exchange chromatography and GlycosepN amide chromatography for the assignment of GU (glucose unit) value. Glycan analysis by HPLC showed that neutral (asialo) oligosaccharide was increased slightly with an increase in osmolality. In portion of sialylated glycan, total relative amount of mono- and di-sialyated glycan was increased but that of tri- and tetra-sialylated glycan decreased as osmolality was increased.

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젓갈 유래 프로바이오틱스의 항산화 및 콜레스테롤 저하 효과

  • Kim, Hyeon-Uk;Lee, Na-Gyeong;Choe, Sin-Yang;Baek, Hyeon-Dong
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.697-700
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    • 2003
  • Lactococcus lactis NK24, Lactobacillus plantarum NK181, and strain HW161 were previously isolated from Jeot-gal. For antioxidative activity, the culture supernatants of there probiotic strains were shown to be 78, 77, and 81% by DPPH method. Also, the peroxidation of linoleic acid of them was investigated and they were about 3.5, 4.6, and 2.0%. And cholesterol-lowering activity of Jeot-gal probiotic strains were about 40, 33, and 60%, respectively.

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Antibacterial Activity and Characteristics of Bacteriocin Produced by Lactobacillus plantarum LMG 7945 (Lactobacillus plantarum LMG 7945가 생산하는 bacteriocin의 항균력과 그 특성)

  • 김상현;이명숙;장동석
    • Journal of Food Hygiene and Safety
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    • v.10 no.2
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    • pp.65-71
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    • 1995
  • Bacteriocins from lactic acid bacteria have attracted much attention in recent years because of their useful worth in increasing safety and extending shelf life of foods. These substances show an inhibitory effect against some food spoilage bacteria and food-borne pathogens. The inhibitory effect fo the bacteriocin produces by lactic acid bacteria against Listeria monocytogenes(L. monocytogenes) was examined in this study. The culture supernatants of 5 kinds of bacteria among the 10 kinds of testes lactic acid bacteria had the inhibitory activity against Listeria sp., various Gram positive and Gram negative bacteria. Bacteriocin produced by Lactobacillus plantarum(Lact. plantarum) LMG 7945 was the most active toward L. monocytogenes. Bacteriocin production of the Lact. plantarum LMG 7945 cultured on MRS broth was increased late logarithmic phase over early stationary phase. This bacteriocin was stable at heat treatment and acidic pH relatively; The activity was retained after heating at 121$^{\circ}C$ for 15min and was active in the pH range of 2~4 but was lost above pH 5.

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Profiling Pyocins and Competitive Growth Advantages of Various Pseudomonas aeruginosa Strains

  • Heo YUN-JEONG;KO KWAN SOO;SONG JAE-HOON;CHO YOU-HEE
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1368-1376
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    • 2005
  • Pseudomonas aeruginosa produces a variety of bacteriocidal substances including pyocins that are active against the same species, but their physiological roles are relatively unknown. Here, we profiled the bacteriocidal activities in the culture supernatants of various P. aeruginosa isolates and describe the competitive growth advantages of strains PAO1 and PA14 over some strains including PAK, which are sensitive to their bacteriocidal activities. These findings suggest that the factors governing the production of pyocins and the resistance to them play important roles in controlling P. aeruginosa populations in its local environments.

The Regulation of Alpha-Amylase Synthesis in Bacillus subtilis

  • Won, Mi-Sun
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.256-260
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    • 1991
  • In B. subtilis, $\alpha$-amylase synthesis is regulated by amyR located directly on the upstream of amyE. Three different amyR alleles have been reported, amyR1, amyR2 and amyR3. Strains bearing the gra-10 mutation which confers derepression for catabolite repression has GlongrightarrowA transition mutation at +5 of amyR1. S1 nuclease mapping demonstrated that transcription initiated at 8 bases downstream from the -10 region of putative E$\sigma^{A}$ promoter P1 in amyR1 and gra-10. In amyR2, the major transcription initiatd at the same place and the minor, 10 bases downstream from -10 of P2. The transcript from P2 contributed approximately 15-20% of total amyE mRNA. S1 nuclease protection experiment indicated that amyE mRNA levels corresponded to the rate of synthesis assumed by specific activities of $\alpha$-amylase in culture supernatants, suggesting that $\alpha$-amylase synthesis is regulated at the level of transcription.n.

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Antitumor Activity of Phytol Identified from Perilla Leaf and its Augmentative Effect on Cellular Immune Response (들깻잎에서 동정한 Phytol의 항암 및 면역활성증강 효과)

  • 김광혁
    • Journal of Nutrition and Health
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    • v.26 no.4
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    • pp.379-389
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    • 1993
  • Several studies have shown that extracts from yellow-green vegetables reveal antitumor activities. In the present study we investigated the effect of phytol in order to elucidate the immunological mechanism of antitumor activity of this substance. The results obtained from the experiment as follows: 1) Phytol showed cytotoxic effect on sarcoma 180 cells in vitro. 2) When phytol was injected into the peritoneal cavity of mice transplanted with sarcoma 180 cells, the average survival time (24.0 days) tended to increase as compared with the nontreated control (19.2 days). 3) When sarcoma 180 cells were injected subcutaneously into the right groin of mice, and then phytol was injected into the peritoneal cavity, the tumor inhibition ratio was 33%. 4) The natural killer(NK) cell activity was significantly augmented by phytol in vitro and in vivo. Similar augmentations of NK cell activity were obtained with culture supernatants of phytol exposed spleen cells and peripheral blood mononuiclear cells. 5) Phytol on the macrophage from peritoneal cavity showed a higher effectiveness in vivo than in vitro. These results indicate that phytol shows the inhibitory effect for growth of sarcoma 180 cells in vitro, also it can augment macrophage and NK cell activities in vivo.

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Size Heterogeneity of Murine Tumor Necrosis Factors Induced from Mouse Peritoneal Macrophages

  • Baik, Na-Gyoung;Jeong, Jee-Yeong;Kim, Soung-Soo
    • BMB Reports
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    • v.28 no.1
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    • pp.46-50
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    • 1995
  • Three kinds of mouse tumor necrosis factor (TNF), which have molecular weights of 35 kDa, 45 kDa, and 18 kDa on SDS-PAGE, were partially purified from serum-free culture supernatants of mouse peritoneal macrophages induced with lipopolysaccharide. Analysis of the native molecular weights by gel filtration indicated that the 18 kDa and 45 kDa TNFs aggregate into 50 kDa and 100 kDa molecules, respectively, while the 35 kDa TNF is contained in high molecular weight aggregates of approximately 200 kDa. The three kinds of cytotoxic factors all elicited tumor reducing responses.

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Isolation and Characterization of Soil Streptomyces Involved in 2,4-Dichlorophenol Oxidation

  • Kang, Min-Jin;Kang, Ja-Kyoung;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.877-880
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    • 1999
  • Over 50 morphologically distinctive soil Streptomyces were isolated from various Jocations in the Yongin area in Korea and visually screened for dye-decoloring activities on an agar plate. Two Streptomyces species (AD001 and ND002) showed strong dye-decoloring activities on the plate containing congo-red and new-fuchin dyes, respectively. Also, the liquid culture supernatants of these species showed 2,4-dicholophenol (DCP) oxidation activities only in the presence of hydrogen peroxide, a characteristic of Actinomycetes lignin-peroxidase (ALiP)-P3 isoform found in dye-degrading S. viridosporus T7A and S. badius 252. Based on their dye-decoloring capabilities and the 2,4-DCP oxidation kinetic data, it is suggested that these Streptomyces secrete not-yet-characterized extracelluar enzyme(s), whose activities are very similar to the ALiP-P3 enzyme.

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Pathogenic effects of porcine reproductive and respiratory syndrome virus isolates in swine tracheal ring culture

  • Park, Bong-kyun;Collins, James E.;Goyal, Sagar M.;Pijoan, Carlos;Joo, Han-soo
    • Korean Journal of Veterinary Research
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    • v.39 no.2
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    • pp.311-317
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    • 1999
  • Pathogenic effects of 29 different porcine reproductive and respiratory syndrome(PRRS) virus isolates were investigated in swine tracheal ring(STR) cultures by examining their effects on the ciliary activity of STR. Inhibition of ciliary movement and destruction of the tracheal epithelium were seen between 72 and 96 hours postinoculation(PI). Virus replication was demonstrated by examining viral infectivity of the supernatants from the STR cultures. PRRS virus antigen in macrophages was detected by a streptavidin-biotin complex(ABC) immunoperoxidase method. Of the 29 PRRS virus isolates, 8 isolates were classified into pathogenic, and the remaining 21 isolates were determined as mildly pathogenic or apathogenic viruses. These results suggest that STR examination may be used as a method for predicting pathogenic variability of PRRS virus isolates.

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Antibody productivity of HBsAg containing both preS2 and S regions expressed in Chinese hamster ovary cells (Chinese hamster ovary세포에서 발현된 pres2 및 S부위 함유 HBsAg의 항체유발능)

  • 정성균;박정민;이상봉;박동우;김동연;김기호;김홍진
    • YAKHAK HOEJI
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    • v.45 no.6
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    • pp.708-714
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    • 2001
  • Many studies have provided evidences that hepatitis B surface antigen (HBsAg) including preS region could be an ideal candidate for a new hepatitis B virus (HBV) vaccine with higher efficacy. We established CHO cell lines, IY-CHO-2 and IY-CHO-11 expressing high levels of HBsAg containing preS2 and S protein by stable transfection method. These cell lines expressed the correct size (about 1 kb in length) of HBsAg mRNA as expected. The purified protein from the culture supernatants of the clones showed the same sizes as those expressed in native hepatitis B virus (24 kDa, 27 kDa, 34 kDa and 36 kDa). Antibody productivity of CHO-derived HBsAg protein at lower dose challenge was higher than the protein containing S region alone expressed in yeast system. These results indicate that CHO-derived HBsAg protein containing preS2 and S region can be effectively used for a better immune response as a HBV vaccine.

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