• Title/Summary/Keyword: extracellular production

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Fucoidan Increases Porcine Neutrophil Extracellular Trap Formation through TNF-α from Peripheral Blood Mononuclear Cells

  • Changwoo Nahm;Yoonhoi Koo;Taesik Yun;Hakhyun Kim;Byeong-Teck Kang;Mhan-Pyo Yang
    • Journal of Veterinary Clinics
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    • v.40 no.3
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    • pp.175-181
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    • 2023
  • Fucoidan extracted from brown seaweed has a variety of biological activities. Neutrophil extracellular traps (NETs) formation is an immune response for the invasion of pathogens. Neutrophils release granule protein and chromatin that form extracellular fibers that bind microbes. These NETs degrade virulence factors and kill bacteria. The aim of this study was to investigate the effect of fucoidan on NET formation of porcine peripheral blood polymorphonuclear cells (PMNs). The NET formation was determined by fluorescence emission of propidium iodide (PI) in PMNs by a fluorescence microplate reader. The production of tumor necrosis factor (TNF)-α from peripheral blood mononuclear cells (PBMCs) was measured by ELISA method. Fucoidan itself did not show any direct effect on NET formation. However, NET formation of PMNs was increased by the culture supernatant from PBMCs treated with fucoidan. The NET formation of PMNs were also enhanced by treatment with recombinant porcine (rp) TNF-α. The ability of culture supernatant from PBMCs treated with fucoidan to increase the NET formation of PMNs was inhibited by addition of goat anti-rp TNF-α polyclonal antibody (pAb) (IgG) prior to the culture. The increase of NET formation by rp TNF-α was also inhibited by goat anti-rp TNF-α pAb (IgG). The level of TNF-α in culture supernatant from PBMCs was increased by treatment with fucoidan. These results suggest that fucoidan increases porcine NET formation, which is mediated by TNF-α produced from PBMCs.

Studies on the Production of Intra- and Extra-cellular Lipids by the Strains in the Genus RHODOTORULA (Rhodotorula 속(屬) 균주(菌株)에 의(依)한 세포(細胞) 내외(內外) 지질생산(脂質生産)에 관(關)한 연구(硏究))

  • Park, Sung-Oh
    • Applied Biological Chemistry
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    • v.17 no.2
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    • pp.93-116
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    • 1974
  • A potent intracellular-lipid-producing yeast, Rhodotorula glutinis var. glutinis SW-17, was screened out from a variety of arable soils, compost heaps, and fodders, and two strains of excellent extracellular-lipid-producing yeasts, Rhodotorula glutinis var. glutinis SW-5 and Rhodotorula graminis SW-54, were screened out from the surface of many species of leaves. And then the intra- and extra-cellular lipid productions by those Rhodotorula yeasts were studied. The results were as follows: 1. During the shaking culture of 8 days at $24^{\circ}C$, both the intra- and extra-cellular lipid accumulation started almost at the stationary phase of growth, when the nitrogen source in the medium was a little more than half used up. The intracellular lipid production by Rhodotorula glutinis var. glutinis SW-17 reached 58.42% (w/w) of dried yeast, and the extracellular lipid production by Rhodotorula graminis SW-54 amounted to 2.62g per liter of the medium. 2. After the carbon and nitrogen sources in the medium were almost consumed, if the yeasts were shake-cultured further in a state of starvation, the yeast cells re-utilized the already produced intra- and extra-cellular lipids and the lipids completely disappeared in the medium in about 90 days. 3. The relative concentration of carbon and nitrogen sources in the media greatly influenced both the intra- and extra-cellular lipid production. When the nitrogen source in the medium was almost used up for the growth of yeast, and excess carbon sources were still available, the lipid production vigorously proceeded. As long as the nitrogen source concentration in the medium was high, the lipid production was greatly suppressed. 4. The optimum pH for both the intra- and extra-cellular lipid production by those yeasts was pH 5.0-6.0. 5. The fatty acid components of the intracellular lipid of Rhodotorula glutinis var. glutinis SW-17 were myristic, palmitic, palmitoleic, stearic, oleic, linoleic, and linolenic acids. The largest components of the fatty acids were palmitic acid equivalent to 30-45% of the whole fatty acids and oleic acid equivalent to 35-50%. 6. The fatty acid components of the extracellular lipid of Rhodotorula glutinis var. glutinis SW-5 and Rhodotorula graminis SW-54 were myristic, palmitic, stearic, oleic, linoleic, linolenic, 3-D-hydroxypalmitic, and 3-D-hydroxystearic acids. The largest components of the fatty acids were 3-D-hydroxypalmitic acid equivalent to 22-25% of the acids and 3-D-hydroxystearic acid equivalent to 13-17%. 7. The polyol component of the intracellular lipids was only glycerol, whereas the polyols of extracellular lipids were glycerol, mannitol, xylitol and arabitol.

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Optimum Culture Conditions for the Production of Fructosyl transferase by Aureobasidium pullulans C-23 (Aureobasidium pullulans C-23 균주에 의한 Fructosyl Transferase의 생산 최적 배양조건)

  • 조원태;임재윤
    • Microbiology and Biotechnology Letters
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    • v.18 no.4
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    • pp.417-422
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    • 1990
  • For optimal production of fructosyl transferase in AureobasidiumpuZZulane C-23, the effect of fermentation conditions for cell growth and fructosyl transferase production were investigated. Sucrose was excellent carbon source. Sucrose concentration for the optimum production of fructosyl transferase was 35%. Enzyme productivity was significantly increased by addition of ammonium oxalate and yeast extract. A time course study for the enzyme production by Aureobasidium pullutans C-23 was carried out. At 2 days incubation, the production of intracellular enzyme was maximum. The extracellular enzyme production was found to be increased up to 6 days.

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Optimal Culture Conditions for the Production of a Novel Extracellular Alkaline Lipase from Yarrowia lipolytica NRRL Y-2178

  • Lee, Geon-Ho;Bae, Jae-Han;Suh, Min-Jung;Kim, Hak-Ryul
    • Journal of Applied Biological Chemistry
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    • v.50 no.2
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    • pp.46-51
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    • 2007
  • Lipases are industrially useful versatile enzymes that catalyze numerous different reactions. Among lipases functioning under extreme conditions, alkaline lipase is useful in detergent industry. Lipase from yeast strain Yarrowia lipolytica NRRL Y-2178 was most active under alkaline condition, and initial medium pH for most lipase production was also alkaline [Lee et al., 2007, J Microbiol Biotechnol, 17(6)]. High lipase production was achieved using Y. lipolytica NRRL Y-2178. Optimal incubation time for lipase production at $25^{\circ}C$ was 72 h. Optimal temperature, when incubated for 72 h, was $27.5^{\circ}C$. Lipase production but not cell growth was very sensitive to concentrations of glucose and glycerol as efficient carbon sources, showing optimal concentrations of 1.0 and 1.5% (w/v), respectively. Lipase production was highly stimulated by $Ca^{2+},\;K^+,\;and\;Na^+$, but was inhibited by $Co^{2+},\;Cu^{2+},\;Mn^{2+},\;Na^+,\;and\;Fe^{2+}$. Maximum lipase production at 0.1 mM $Ca^{2+}$ for 72 h incubation at $27.5^{\circ}C$ was 649 units/mL.

Typha orientalis inhibits inflammatory cytokine expression through suppression of ERK phosphorylation in HMC-1 cells

  • Choi, In-Young;Na, Ho-Jeong;Um, Jae-Young;Kim, Hyung-Min;Hong, Seung-Heon;Sim, Kuk-Jin;Song, Bong-Keun;Nam, Gi-Hye;Choung, Se-Young;Jeong, Hyun-Ja
    • Advances in Traditional Medicine
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    • v.10 no.1
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    • pp.7-12
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    • 2010
  • Typha orientalis' stem (TOS) is traditionally used as an herbal medicine for difficulty in urination, galactophoritis purulenta, whooping cough, and allergic dermatitis. However, its effect in experimental models remains unknown. Here, we report the effect of TOS on the phorbol 12-myristate 13-acetate (PMA) plus calcium ionophore A23187-induced inflammatory cytokine production and extracellular signal-regulated kinase (ERK) activation in the human mast cell line, HMC-1. TOS inhibited PMA plus A23187-induced cytokines such as tumor necrosis factor-alpha (TNF-$\alpha$) and interleukin (IL)-6. Maximal inhibition rate of TNF-$\alpha$ and IL-6 production by TOS (1 mg/ml) was about 44.02%, and 45.20%, respectively (P < 0.05). In addition, TOS inhibited the expression of TNF-$\alpha$ and IL-6 mRNA under the same condition. Moreover, TOS partially blocked PMA plus A23187-induced ERK phosphorylation. These results suggested TOS could inhibit the cytokine production through blocking of ERK activity.

Isolation and Characterization of Pretense Producing Bacteria from Soil (토양으로부터 Protease 생산 세균의 분리 및 특성)

  • 김관필;이창호;우철주;김남형;배동호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.754-759
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    • 2002
  • In order to develope a new pretense applicable to industries, a bacterium which produces a remarkable amount of extracellular pretense were isolated from soil. About 10 bacterial strains producing pretense were isolated from samples of soil, and strain PANH765 showed the highest activity of pretense production among them. The strain was identified as Bacillus subtilis according to the Bergey's Manual of Systematic Bacteriology based on its morphological, cultural and physiological characteristics. B. subtilis PANH765 showed the maximal production of pro-tease in the medium containing 2.0% glucose, 1.0% yeast extract, 0.2% ammonium nitrate, 0.02% ferrous sulfate and 0.02% dipotassium hydrogen phosphate. Under the optimal condition with temperature of 3$0^{\circ}C$, initial pH of 7.0 and shaking speed of 150 rpm, the pretense production reached a maximum level with 36 hr cultivation (6.34 U).

Isolation and Enzyme Production of a Mannanase-producing Strain, Bacillus sp. WL-3. (Mannanase를 생산하는 Bacillus sp. WL-3 균주의 분리와 효소 생산성)

  • 오영필;이정민;조기행;윤기홍
    • Microbiology and Biotechnology Letters
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    • v.30 no.3
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    • pp.247-252
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    • 2002
  • A bacterium producing the extracellular mannanase was isolated from Korean formented food and has been identified as a member of the genus Bacillus from the result of the phylogenic analysis based on partial 165 rRNA sequences. The isolate, named Bacillus sp. WL-3, was shown to be similar to B. subtilis strain on the basis of its biochemical properties. The mannanase of culture supematant was the most active at $55^{\circ}C$ and pH 6.0. The additional carbohydrates including u-cellulose, avicel, oat spelt xylan, guar gum and locust bean gum (LBG) increased the mannanase productivity. Especially, the maximum mannanase productivity was reached 65.5 U/ml in LB medium supplemented with 0.5% (w/v) LBG, which was 131-folds more than that in LB medium. It was sug-gested that the increase of mannanase production was owing to induction of mannanase biosynthesis by LBG hydrolysates transported following initial hydrolysis by extracellular mannanase during the cell growth. The molec-ular weight of WL-3 mannanase was estimated to approximately 38.0 kDa by zymogram on SDS-PAGE.

Panax Ginseng inhibited HIF-1a activation and inflammatory cytokine in HMC-1 cells activated by phorbol myristate acetate and A23187

  • Choi, In-Young;Jeong, Hyun-Ja;An, Hyo-Jin;Kang, Tae-Hee;Zo, Chul-Won;Song, Bong-Keun;Park, Eun-Jeong;Kim, Eun-Cheol;Um, Jae-Young;Kim, Hyung-Min;Hong, Seung-Heon
    • Advances in Traditional Medicine
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    • v.8 no.4
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    • pp.440-447
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    • 2008
  • This study investigated the role of Panax ginseng (PG) on the phorbol myristate acetate (PMA) + calcium ionophore A23187-induced hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$) activation, phosphorylation of the extracellular signal-regulated kinase (ERK), and inflammatory cytokine production from the human mast cell line, HMC-1. HIF-$1{\alpha}$ and phosphorylation of ERK were observed by Western blotting. The inflammatory cytokine production was determined by enzyme-linked immunosorbent assay. PG inhibited the PMA+A23187-induced HIF-$1{\alpha}$ expression and the subsequent production of vascular endothelial growth factor. In addition, PG suppressed PMA + A23187-induced phosphorylation of ERK. We also show that the increased cytokines interleukin (IL)-$1{\beta}$, IL-6, and tumour necrosis factor-${\alpha}$ level was significantly inhibited by treatment of PG. In the present study, we report for the first time that PG is an inhibitor of HIF-$1{\alpha}$ and cytokines on the mast cell-mediated inflammatory responses.

Control of Catabolite Repression by Limit Feed of Cellobiose in Cellulomomas sp. (Cellulomonas sp.에 있어서 셀로비오스의 미량공급에 의한 생성물 저해의 조절)

  • Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.3 no.2
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    • pp.235-243
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    • 1976
  • As an investigation on the catabolite repression system in cellulase production by Cellulomons sp. CS1-1, the organism was tested on the avicel overlay plates containing glucose or cellobiose at a range of concentration and was grown in continuous culture vessel, supplied by cellobiose medium, aiming the enhanced production of extracellular CM-cellulase at low dilution rates. Product inhibition of cellulase action by cellobiose was also tested. The results obtained are: i) no inhibition of CM-cellulase was observed up to 10 mM(3.4mg/ml) cellobiose in the reaction mixture, however 30% inhibition was observed at 20mM and 55% at 50mM, ii) the tests of catabolite repression on the solid media were successful, and avicel degradation was markedly repressed by glucose or cellobiose, iii) at low concentrations of cellobiose, dilution rate 0.05 and $1.0hour^{-1}$, no significant increase was observed in the production of either intra or extracellular CM-cellulase.

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Production and Characterization of Antifungal Chitinase of Bacillus licheniformis Isolated from Yellow Loess (황토로부터 분리한 Bacillus licheniformis의 항진균 chitinase 생산과 효소 특성)

  • Han, Gui Hwan;Bong, Ki Moon;Kim, Jong Min;Kim, Pyoung Il;Kim, Si Wouk
    • KSBB Journal
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    • v.29 no.3
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    • pp.131-138
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    • 2014
  • In this study, we isolated two novel chitinase producing bacterial strains from yellow loess samples collected from Jullanamdo province. The chitinase producing bacteria were isolated based on the zone size of clearance in the chitin agar plates. Both of them were gram positive, rod ($2{\sim}3{\times}0.3{\sim}0.4{\mu}m$), spore-forming, and motility positive. They were facultative anaerobic, catalase positive and hydrolyzed starch, gelatin, and casein. From the 16s rRNA gene sequence analysis, the isolates were labeled as Bacillus licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02. The isolates showed higher extracellular chitinase activities than B. licheniformis ATCC 14580 as a control. The optimum temperature and pH for chitinase production were $40^{\circ}C$ and pH 7.0, respectively. Response Surface Methodology (RSM) was used to optimize the culture medium for efficient production of the chitinase. Under this optimal condition, 1.5 times higher chitinase activity of B. licheniformis KYLS-CU02 was obtained. Extracellular chitinases of the two isolates were purified through ammonium sulfate precipitation and anion-exchange DEAE-cellulose column chromatography. The specific activities of purified chitinase from B. licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02 were 7.65 and 5.21 U/mg protein, respectively. The molecular weights of the two purified chitinases were 59 kDa. Further, the purified chitinase of B. licheniformis KYLS-CU01 showed high antifungal activity against Fusarium sp.. In conclusion, these two bacterial isolates can be used as a biopesticide to control pathogenic fungi.