• Title/Summary/Keyword: $MgB_4$ addition

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Purification and Characterization of a Novel Salt-tolerant Protease Produced by Saccharomyces sp. B101 Isolated from Baker's Dough Yeast

  • Hwang, Joo-Yeon;Kim, Sang-Moo;Heo, Seok;Kim, Cheon-Jei;Lee, Chi-Ho;Lee, Si-Kyung
    • Food Science and Biotechnology
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    • v.17 no.4
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    • pp.766-771
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    • 2008
  • The proteolytic enzyme from Saccharomyces sp. B101 was purified to homogeneity by ammonium sulfate fractionation, ultrafiltration, diethyl aminoethyl (DEAE)-Sephadex A-50 ion-exchange chromatography, and Sephadex G-100 gel filtration chromatography from the culture supernatant of Saccharomyces sp. B101. The specific activity and the purification fold of the purified enzyme were 4,688.9 unit/mg and 18, respectively. The molecular weight of the purified enzyme was estimated to be 33 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH and temperature for the enzyme activity were pH 8.5 and $30^{\circ}C$, respectively. The enzyme activity was relatively stable in the pH range of 6.5-8.5 at below $35^{\circ}C$. The salt-tolerance and stability for the enzyme activity were relatively stable even at NaCl concentrations of 10 and 15%. The activity of enzyme was inhibited by $Ag^{2+}$ and $Fe^{2+}$, and activated by $Mn^{2+}$. In addition, the enzyme activity was potently inhibited by ethylenediaminetetraacetic acid (EDTA) and phenylmethyl sulfonylfluoride (PMSF). Based on these findings we concluded that the purified enzyme was a serine protease. Km and Vmax values for hammastein milk casein were 1.02 mg/mL and 278.38 unit/mL, respectively.

Healing and preventive effects of low-esterified pectin on liver injury induced by carbon tetrachloride in rats

  • Khotimchenko, Yuri S.;Kolenchenko, Elena A.;Khotimchenko, Maxim Y.;Kovalev, Valeri V.
    • Advances in Traditional Medicine
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    • v.4 no.1
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    • pp.28-36
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    • 2004
  • The purpose of this study was to investigate the pharmacological effects of low-esterified pectin on carbon tetrachloride $(CCL_4)-induced$ hepatotoxicity in rats. The study included two experiments. In the first experiment the animals were given daily $CCL_4$ through gavage for 7 days and then 10, 50, or 250 mg/kg b.w. of pectin for 21 days. At the end of experiment rats were killed within 24 hours. The increased bilirubin level, enhanced alanine aminotransferase and aspartate aminotransferase activity in plasma induced by $CCL_4$ were partly normalized by pectin administration in a dose-dependent manner. The pectin treatment also resulted in significant recovery of $CCL_4-induced$ decrease of the liver glycogen content. In addition, pectin significantly improved $CCL_4-induced$ alterations of pro-oxidant and antioxidant biochemical parameters in liver and plasma compared to those of rats administered $CCL_4$. In the second experiment the animals were given daily 10, 50 or 250 mg/ kg b.w. of pectin for 21 days before a 7-day administration of $CCL_4$. Rats were killed 24 hours after the end of experiment. Pretreatment with pectin before $CCL_4$ administration resulted in significantly inhibited increase of the blood enzymatic activities of alanine and aspartate aminotransferases and bilirubin level in a dose-dependent manner. Also, preliminary administration of pectin prevented elevation of malondialdehyde and conjugated diene levels in liver and plasma as well as a reduction of glutathione content in liver of rats given $CCL_4$. These results suggest that low-esterified pectin exert healing and preventive effects on $CCL_4-induced$ hepatotoxicity in rats.

Anti-inflammatory Effects of the Aqueous Extract of Hwangnyeonhaedok-tang in LPS-activated Macrophage Cells (LPS로 활성화된 대식세포에서 황련해독탕(黃連解毒湯) 물추출물의 염증매개물질 억제효과)

  • Kim, Dae-Hee;Park, Sook-Jahr;Jung, Ji-Yoon;Kim, Sang-Chan;Byun, Sung-Hui
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.39-47
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    • 2009
  • Objectives : Hwangnyeonhaedok-tang (Huanglian Jiedu Tang; HHT) has been widely used for purging' 'fire' and lessening virulence of any pathogenic organism. However it has been rarely conducted to evaluate the immuno-biological activity. In this study, we evaluated anti-inflammatory effects of HHT in LPS-activated Raw264.7 cells. Methods : Cells were treated with $1\;{\mu}g/ml$ of LPS 1 h prior to the addition of HHT. Cell viability was measured by MTT assay. The production of NO was determined by reacting cultured medium with Griess reagent. PGE2 and proinflammatory cytokines were detected by ELISA. Expression of iNOS, COX-2, $I{\kappa}B{\alpha}$ and NF-${\kappa}B$ were analyzed by immunoblot analysis. Results : All three doses of HHT (0.03, 0.10 and 0.30 mg/ml) had no significant cytotoxicity during the entire experimental period. The levels of NO and PGE2 were dramatically augmented by LPS compared to control. However, HHT extract dose-dependently reduced these increases. Expression of iNOS and COX-2 protein were also decreased by treatment with HHT extract. Furthermore, HHT extract significantly reduced the nuclear translocation of NF-${\kappa}B$ which is critical in regulating inflammation through transcription of iNOS and COX-2. In addition, HHT extract reduced the elevated production of inflammatory cytokines including TNF-$\alpha$, IL-$1{\beta}$ and IL-6. Conclusions : The results in this study demonstrate that HHT extract exerts anti-inflammatory activities through the inhibition of NO, PGE2 and proinflammatory cytokines production via the suppression of NF-${\kappa}B$.

DA-9201 Shows Anti-Asthmatic Effects by Suppressing NF-$\kappa$B Expression in an Ovalbumin-Induced Mouse Model of Asthma

  • Lee Seung-Ho;Seo Mi Jung;Choi Seul Min;Sohn Yong Sung;Kang Kyung Koo;Ahn Byoung Ok;Kwon Jong Won;Yoo Moohi
    • Archives of Pharmacal Research
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    • v.28 no.12
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    • pp.1350-1357
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    • 2005
  • Nuclear factor kappa B (NF-$\kappa$B) regulates the expression of multiple cytokines, chemokines, and cell adhesion molecules that are involved in the pathogenesis of asthma. We investigated the anti-asthmatic effects and the mechanism of action of DA-9201, an extract of the black rice, in a mouse model of asthma. Mice immunized with ovalbumin (OVA) were administered with DA-9201 (30, 100 or 300 mg/kg) or dexamethasone (DEXA, 3 mg/kg) for 2 weeks and challenged with aerosolized OVA during the last 3 days. Anti-asthmatic effects were assessed by means of enhanced pauses, level of total lgE and Th2 cytokines in plasma or bronchoalveolar lavage fluid (BALF), the percentage of eosinophils in BALF, and histopathological examination. The expression of NF-$\kappa$B in nuclear and cytoplasmic fraction and its DNA-binding activity in lung tissues were analyzed by means of Western blotting and electrophoretic gel mobility shift assay (EMSA), respectively. DA-9201 significantly reduced airway hyperrespon-siveness (AHR), total lgE level in plasma and BALF, IL-4, IL-5, and IL-13 levels in BALF, and the percentage of eosinophils in BALF. Tissue inflammation was significantly improved by DA­9201 treatment. In addition, DA-9201 dramatically suppressed the expression of NF-$\kappa$B and its DNA-binding activity. These results suggest that DA-9201 may be useful for the treatment of asthma and its efficacy is related to suppression of NF-$\kappa$B pathway.

The Effect of Solid Solution Heat-Treatment Temperature on the Tensile Property in Intermediate Thermo-Mechanical Treated Al-Li Alloys (중간가공열처리한 AI-Li계 합금의 인장성질에 미치는 용체화처리온도의 영향)

  • Yoo, C.Y.;Lee, K.B.
    • Journal of the Korean Society for Heat Treatment
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    • v.4 no.1
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    • pp.37-41
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    • 1991
  • In this study, the effect of solution treatment temperature on the tensile property in intermediate thermo-mechanical treated Al-Li alloys are investigated. After the intermediate thermo-mechanical treated Al-Li, Al-Li-Mg and Al-Li-Mg-Zr alloys were solution treated at various temperatures (500, 520 and $540^{\circ}C$), these were aged at $190^{\circ}C$, $240^{\circ}C$ and tested tensile properties. The results obtained from the experiment are as follows ; 1) The optimum solution heat-treatment temperature is $540^{\circ}C$ for a Al-Li alloy, and the recrystallized grain size is about $70{\mu}m$. 2) The optimum solution heat-treatment temperature is $500^{\circ}C$ for a Al-Li-Mg alloy, and the recrystallized grain size is the most coarse in all alloys. 3) The tensile property is independent of the solution treatment temperature in a Al-Li-Mg-Zr alloy, and the recrystallized grain size is the finest owing to addition of Zr.

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Quality Characteristics of Fermented Mate(ILex paraguarensis) Leaf Tea (Mate (ILex paraguarensis) 발효차의 품질특성)

  • Hong, Joo-Heon
    • Food Science and Preservation
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    • v.17 no.4
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    • pp.500-506
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    • 2010
  • This study was conducted to compare and analyze the active and general components in mate leaf tea according to degree of fermentation conditions. The contents of tannic acid of hot water extracts from #1 (mate leaf), #2 (Mate leaf after fermentation and roasting), #3 (Mate leaf after Pan-firing), and #4 (Mate leaf after final drying) were decreased according to degree of fermentation conditions. Polyphenol contents of hot water extracts were approximate in #1 and #3, with measurements of 43.45 mg/g and 38.20 mg/g, respectively. Caffeine contents were 6.78 mg/g in #1, 4.30 mg/g in #3, and 3.65 mg/g in #4. In addition, the level of total free amino acid of #1 was higher than that of #2, #3, and #4. Lightness (L) and Redness (a) values increased and yellowness (b) values decreased after fermentation. When sensory tests were conducted, mate leaf tea after fermentation had pleasant taste.

Gene cloning of β-mannanase C from Cellulosimicrobium sp. YB-43 and characterization of the enzyme (Cellulosimicrobium sp. YB-43으로부터 mannanase C 유전자의 클로닝과 효소 특성)

  • Yoon, Ki-Hong
    • Korean Journal of Microbiology
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    • v.54 no.2
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    • pp.126-135
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    • 2018
  • The characteristics of enzyme and gene for mannanase B had been reported from Cellulosimicrobium sp. YB-43 producing some kind of mannanase. A gene coding for the enzyme, named mannanase C (ManC), was expected to be located downstream of the manB gene. The manC gene was cloned by polymerase chain reaction and sequenced completely. From this nucleotide sequence, ManC was identified to consist of 448 amino residues and contain a carbohydrate binding domain CBM2 besides a catalytic domain, which was homologous to mannanase belonging to the glycosyl hydrolase family 5. The catalytic domain of ManC showed the highest amino acid sequence similarity of 55% with the mannanases from Streptomyces sp. SirexAA-E (55.8%; 4FK9_A) and S. thermoluteus (57.6%; BAM62868). The His-tagged ManC (HtManC) lacking N-terminal signal peptide with hexahistidine at C-terminus was produced and purified from cell extract of recombinant Escherichia coli. The purified HtManC showed maximal activity at $65^{\circ}C$ and pH 7.5, with no significant change in its activity at pH range from 7.5 to 10. HtManC showed more active on konjac and locust bean gum (LBG) than guar gum and ivory nut mannan (ivory nut). Vmax and Km values of the HtManC for LBG were 68 U/mg and 0.45 mg/ml on the optimal condition, respectively. Mannobiose and mannotriose were observed on TLC as major products resulting from the HtManC hydrolysis of mannooligosacharides. In addition, mannobiose and mannose were commonly detected as the hydrolyzed products of LBG, konjac and ivory nut.

Quality of Persimmon Jelly by Various Ratio of Dried Persimmon Extract (곶감추출물 첨가비율에 따른 곶감젤리의 품질)

  • Kim, Jun-Han;Kim, Jong-Kuk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.7
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    • pp.1091-1097
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    • 2005
  • This study was carried out to develop jelly food using dried persimmon. Jelly products were prepared with the ratio of 5, 10, 15, 20 and $25\%$ of dried persimmon extract. Quality characteristics of its products were investigated. Proximate compositions of jelly were $6.74\~14.03\%$ of moisture, $6.83\~7.53\%$ of crude protein, $0.62\~1.16\%$ of crude lipid, $2.61\~13.21\%$ of crude ash, respectively. Water activity and brix $(\%)$ of jelly products ranged from 0.678 and $56.66\%\;to\; 0.748\;and\;76.59\%$. The pH and total acidity of jelly products ranged from 5.30 and $0.06\%$ to 5.38 and $0.09\%$. In the Hunter's color values, L, a and b vaules of jelly products were increased, respectively. Major free-sugar and organic acid of jelly products were maltose ($753\~1,297mg/100g$) and malic acid (263mg/100g in $25\%$ dried persimmon jelly). Major mineral of jelly products was K ($69.64\~154.37mg/100g$). In the texture property, addition of dried persimmon extract decreased gumminess and chewiness. In sensory score of dried persimmon jelly, color, flavor, texture and sweetness of $15\%$ dried persimmon jelly were high score, taste and overall acceptance of jelly products were high score, respectively. Judging from research results of the jelly products, recommended substitution level for addition of dried persimmon extract in jelly was $10\~15\%$.

The Horse Manure Fertilizers and Pollutants Characteristics Analysis Research (말 분뇨의 비료성분 및 오염물질 특성 조사 연구)

  • Kwag, J.H.;Jeong, M.W.;Choi, D.Y.;Kim, J.H.;Kim, Y.H.;Park, J.C.;Kim, I.C.;Yang, C.B.;La, C.C.
    • Journal of Animal Environmental Science
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    • v.18 no.3
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    • pp.201-206
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    • 2012
  • Horse manure general composition and contaminants, heavy metals content analysis was investigated as follows. The moisture content of horse feces was 76.1% and 96% for horse urine. The average T-N, T-P, T-K concentrations of the racing horse were 0.18%, 0.10%, 0.22% respectively and 2.88%, 0.0015%, 0.84% for urine. And the average BODs, COD, SS concentrations were 26,906 $mg/{\ell}$, 36,642 $mg/{\ell}$, 89,375 $mg/{\ell}$ respectively and 14,298 $mg/{\ell}$, 7,484 $mg/{\ell}$, 6,987 $mg/{\ell}$ for urine. In addition, the Cr, Cu, Ni, Cd, Pb, Zn concentrations of feces were 2.65 mg/kg, 7.05 mg/kg, 1.24 mg/kg, 0.07 mg/kg, 0.01 mg/kg, 45.11 mg/kg respectively and 1.52 mg/kg, 1.62 mg/kg, 0.00 mg/kg, 0.03 mg/kg, 0.03 mg/kg, 4.49 mg/kg for urine on the fresh matter basis. The end of the survey was that the quality of the compost can be used as a raw material, and manure is necessary to constantly driven forward horse manure composting research.

Immunostimulating Effect of Palmiboshinwhan (baweibushenwan) Extracts in Methotrexate-induced Immunosuppressed SD rats. (팔미보신환(八味補腎丸)이 Methotrexate로 유발(誘發)된 SD Rat의 면역기능저하(免疫機能低下)에 미치는 영향(影響))

  • Kang, Seok-Bong
    • The Journal of Korean Medicine
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    • v.29 no.3
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    • pp.21-37
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    • 2008
  • Objectives: The purpose of this study was to verify the effect of Palmiboshinwhan (PMBSW) in methotrexate (MTX)-induced immunosuppressed SD rats. Methods: The test articles were once a day dosed for 14 days by gastric gavage from 2 days after last MTX-dosing, and changes in body weight, spleen weight and total blood leukocyte numbers were observed with total lymphocyte numbers, B and T lymphocyte percentages, CD3+CD4+, CD3+/CD8+, CD4+/CD8+ T lymphocyte percentages in the blood and spleen, the serum interleukin (IL)-2 levels and the productivity of IL-2 of splenic cells. Result & Conclusion: It is concluded that PMBSW has relatively good immunostimulating effect in the MTX-induced immunosuppressed SD rats. Theefficient dosage was considered above 500mg/kg. In addition, it is considered that the immunostimulating effect of PMBSW was mediated to both the B and T lymphocytes. The more favorable effects were detected in T lymphocytes rather than B lymphocytes, and PMBSW showedrelatively good stimulating potential against CD4+ T lymphocytes but not any stimulating effect against CD8+ T lymphocytes in the present study.

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