• Title/Summary/Keyword: $MgB_4O_7$

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Enzymatic Properties of Cytidine Deaminase Encoded by cdd Gene in Bacillus subtilis (Bacillus subtilis의 cdd 유전자에 의해 코드되는 Cytidine Deaminase의 효소학적 성질)

  • Song, Bang-Ho;Yoon, Mi-Sook;Kim, Kyung-Hwa;Yeo, Jeung-Sook;Jan Neuhard
    • Microbiology and Biotechnology Letters
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    • v.16 no.6
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    • pp.468-475
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    • 1988
  • The cloned B. subtilis cdd gene encoding cytidine/2'deoxycytidine deaminase (EC 3.5.4.5) was expressed in the cdd deficient B. subtilis mutant ED40. The gene was isolted from the cdd complementing plasmid pSO21, and inserted into the EcoR1/Pvu1 sites of pGB215-110 ΔB, which is a temperature sensitivie E. coli-B. subtilis shuttle vector. In the transformed B. subtilis ED4O harboring the resulting plasmid pSO100, cdd was expressed at several hundred fold elevated levels, and the cytidine deaminase activity in E. coli containing pSO100 was twice the level in B. subtilis/pSO0100. The Km value for cytidine of the partially purified enzyme is 1.88$\times$10$^{-4}$M at pH 7.0 and the V$_{max}$ = 11.1 $\mu$mol/min/mg of protein. The enzyme was completely inhibited by 0.1M mercaptoethanol and HgCl$_2$. The inhibition by p-chrolomercurybenzoic acid showed a Ki = 5 uM. These results suggest that sulfhydryl reagents block an active site thiol group, and/or disturb the formation of the tetrameic holoenzyme.

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Properties of the CMCase produced by Pseudomonas sp. YD-15 (Pseudomonas sp. YD-15가 생산하는 CMCase의 특성)

  • Lee, Jeong-Woo;Kim, Chang-Nam;Hur, Nam-Yun;Oh, Doo-Hwan
    • Applied Biological Chemistry
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    • v.35 no.3
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    • pp.173-178
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    • 1992
  • A bacterium having CMCase activity was isolated form soil and identifed as a Pseudomonas sp YD-15. The optimum conditions for the production of CMCase were avicel 1.2%, yeast extract 0.5%, $KNO_3$ 0.06%, $K_2HPO_4$ 0.2%, $MgSO_4$ $7H_2O$ 0.15%, pH 8.0, $30^{\circ}C$ and 60 hours cultivation. The CMCase was purified 15.2 folds with 14ft yield through ammonium sulfate precipitation, DEAE-sepharose column chromatography and sephadex G-100 gel filtration chromatography. The optimum pH and temperature for the enzyme activity were 6.0 and $50^{\circ}C$, respectively. The enzyme was stable between pH 5.0 and 8.0, below $50^{\circ}C$. The molecular weight was calculated about 100,000 by SDS-polyacrylamide gel electrophoresis. $K_m$ value for CMC used as a substrate was 40 mg/ml.

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Chitinase of Multifunctional Antagonistic Bacterium Bacillus amyloliquefaciens 7079 against Phy-tophathogenic fungi (식물병원진균을 길항하는 chitinase 생산성 생물방제균 Bacillus amyloliquefaciens 7079의 선발과 chitinase 생산조건)

  • 한옥경
    • Microbiology and Biotechnology Letters
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    • v.29 no.3
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    • pp.142-148
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    • 2001
  • An indigenous antagonistic bacterium Bacillus sp. 7079 was isolated from a local soil sampled at Kyongju area in Korea . The strain has strong antagonistic ability which was originated from multifunctional mechanisms of chitinase and antibiotic and is a powerful antagonistic biocontrol agent against red-pepper rotting fungus Phytophthora capsici and Wilt fungus Fusarium oxysporum. The chitinase might degrade the cell wasll for Fusarium species. The selected Bacilus sp. 7079 was identified as a Bacillus amyloliquefaciens 7079. The maximal production of the chitinase from B, amyloliquefaciens 7079 were obtained in chitin-yeast extract medium containing 0.7%, $K_2$$HPO_4$, $0.2KH_2$$PO_4$, 0.1% ($NH_4$)$_2$$SO_4$, 0.05% sodium cirate, 0.01% $MgSO_4$$7H_2$O, 0.1% yeast extract and 0.1% colloidal chitin after cultivation of 3 days at pH 7.0 and $30^{\circ}C$. The best carbon and nitrogen sources for the production of the chitinase from B amyloliquefaciens 7079 were determined to be 0.1% colloi- dal chitin and 0.15% proteose peptone NO 3 respectively, The antagonistic activity of B amyloliquefaciens 7079 was confirmed using P. capsici by in vivo pot test with red-pepper plant.

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Development of new substrate using redginseng marc for bottle culture of oyster mushroom (Pleurotus ostreatus) (홍삼박을 이용한 병재배 느타리버섯의 첨가배지 개발)

  • Lee, Chan-Jung;Han, Hye-Su;Jhune, Chang-Sung;Cheong, Jong-Chun;Oh, Jin-A;Kong, Won-Sik;Park, Gi-Chun;Park, Chun-Geon;Shin, Yu-Su
    • Journal of Mushroom
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    • v.9 no.4
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    • pp.139-144
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    • 2011
  • This study was carried out to investigated mixing ratio of redginseng marc using organic waste resource for production cost-reduction of oyster mushroom. Redginseng marc were examined as substitute of cottonseed meal which was primary nutritive material of mushroom growing substrate. Total nitrogen and carbon source of redginseng marc was 45% and 2.7%, respectively and C/N ratio was 16.7. Total nitrogen source and pH of substrate mixed with redginseng marc was 4.6~4.9 and 2.5~2.8, respectively. The contents of $P_2O_5$, $K_2O$ and MgO were decreased by increasing redginseng marc, but increased at the 20% redginseng marc. The contents of CaO was increased by increasing redginseng marc, but there was no significant difference in $Na_2O$ content. The more mixing ratio of redginseng marc was increased in column test, the more mycelial growth growed slowly. Yields of fruiting body was no significant difference compared with control group by adding of 10~50% redsingeng marc instead of cottonseed meal, but decreased sharply at only 20% redsingeng marc without cottonseed meal. The L value of pileus by increasing redginseng marc decreased during mushroom harvest, but there was no significant difference in the a-value and the b-value.

Effect of Protein Supplementation, O2 Concentration and Co-Culture on the Development of Embryos Produced by Nuclear Transfer Using Cultured Cumulus Cells in Hanwoo (Korean Cattle)

  • Im, G.S.;Yang, B.S.;Park, S.J.;Im, S.K.;Yang, B.C.;Yi, Y.J.;Park, C.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.9
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    • pp.1260-1266
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    • 2001
  • The effect of protein supplementation, $O_2$ concentration and co-culture on the development of embryos produced by nuclear transfer using cultured cumulus cell was investigated. Recipient oocytes and cumulus cells were obtained from the ovaries of the slaughtered Hanwoo cows. Donor cumulus cells were cultured in Dulbecco's modified Eagle medium containing 10% fetal bovine serum at 5% $CO_2$ in air at $38.5^{\circ}C$. The 1 to 6 passages of cumulus cells were isolated and used as donor cells. The in vitro matured oocytes were enucleated and then the isolated donor cells were introduced. One $15{\mu}s$ pulse of 180 volts was applied to induce the fusion between karyoplast and cytoplast. The fused embryos were activated with $10{\mu}M$ calcium ionophore for 5 min and 2 mM 6-dimethylaminopurine for 3 h. To examine the effect of protein supplementation, nuclear transfer (NT) embryos were cultured in one of the following 4 treatments : 1) CR1aa + 3 mg/ml BSA for 7 days ; 2) CR1aa + 10% FBS for 7 days ; 3) CR1aa + 1.5 mg/ml BSA + 5% FBS for 7 days ; and 4) CR1aa + 3 mg/ml BSA for first 3 days and then CR1aa + 1.5 mg/ml BSA + 5% FBS for 4 days. Culture took place at 5% $CO_2$, 5% $O_2$ and 90% $N_2$ at $38.5^{\circ}C$. Although there were no significant differences in cleavage rate among different protein supplements, the rates of blastocyst formation were significantly different. When NT embryos were cultured in the medium supplemented with only BSA, they could develop to only morula not to blastocyst. However, when FBS was supplemented, NT embryos developed to blastocyst stage. In order to investigate the effect of $O_2$ concentration and co-culture, NT embryos were cultured in CR1aa + 1.5 mg/ml BSA + 5% FBS with or without cumulus cell co-culture at an atmosphere of 5% $CO_2$ in air (20% $O_2$) or 5% $CO_2$, 5% $O_2$, 90% $N_2$ (5% $O_2$) at $38.5^{\circ}C$ for 7 days. The percentage of blastocyst development was significantly higher when the NT embryos were cultured at an atmosphere of 5% $O_2$ than that of 20% $O_2$ (p<0.05). However, there was no significant difference between with and without cumulus cell co-culture at an atmosphere of 5% $O_2$ or 20% $O_2$. Fifty embryos were transferred to 25 recipients and 5 recipients were pregnant at 100 days. From 5 pregnant cows, only one cow was delivered of female twin. In conclusion, the embryos reconstructed by enucleation of metaphase II oocytes and introduction of the cycling and quiescent cumulus donor cells in Hanwoo had developmental potential to term after embryo transfer to recipient cows.

Effect of Ascorbic Acid and Kimchi on the Metabolism of Xenobiotics in Guinea Pigs Administered Polycyclic Aromatic Hydrocarbon (Ascorbic Acid 및 김치가 다환 방향족 탄화수소 섭취 Guinea Pigs 의생체 이물 대사에 미치는 영향)

  • 유리나;홍순명;최석영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.19 no.5
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    • pp.403-410
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    • 1990
  • The effect of ascorbic acid (AsA) or kimchi on the metabolism of xenobiotics in guinea pig administered policyclic aromatic hydrocarbons(PAH) an environmental pollutant was inves-tigated. Guinea pigs given oral supplementation were divided into 4 groups. Group A(control) was supplemented with 2mg of AsA/day group B with 2mg of AsA and PAH/day gruop C with 50mg of AsA and PAH/day. Changes of liver cytochrome P-450 contents and 7-ethoxycou-marin O-deethylase(ECOD) activity glutamate-oxaloacetate transferase(GOT) activity in serum and AsA contents in various tissues were determined. Cytochrome P-450 contents ECOD activity and GOT activity increased in B, C and D groups administered PAH. Moreover cytoch-rome P=450 contents and ECOD activity in group C were higher than those of group A and D. The content of AsA in the tissues of group B was lower than that of A, C and D groups respectively. These results suggested that AsA was necessary in the PAH metabolism moreo-ver supplementation of large amount of AsA or kimchi in the guinea administered PAH had an influence in the drug-metabolizing enzyme induction thus caused a decrease of AsA in tissues.

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Flavonoids of Gomphocarpus sinaicus and Evaluation of Some Pharmacological Activities

  • Batran, Seham A. El;Abdel-Azim, Nahla S.;Abdel-Shafeek, Khaled A.;Shahat, Abd-Elatty A.;El-Missiry, Moustafa M.
    • Natural Product Sciences
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    • v.11 no.4
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    • pp.233-239
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    • 2005
  • The aerial parts of Gomphocarpus sinaicus Boiss. yielded four flavonoids that were identified as isorhamnetin 3-O-rhamnoglucoside (1), luteolin-7-O-glucoside-3-O-rhamnoside (2), rutin (3) and rutin-7-O-rhamnoside (4). All of the isolated flavonoids were identified by spectroscopic methods (UV, FAB-MS, $^1H-NMR\;&\;^{13}C-NMR$) and in comparison with the literature data. The isolated flavonoids 1, 2 and 4 are reported here for the first time from Gomphocarpus sinaicus Boiss. Three sets of experiments were carried out using the defatted alcoholic extract of Gomphocarpus sinaicus Boiss: the $1^{st}$ experiment indicated that the $LD_{50}$ was 49.82 mg/100 g b.wt. of intraperitoneally (i.p.) injected mice. The toxic signs were recorded within the first 24 hr post-injection. The $2^{nd}$ experiment revealed that the extract of the plant exhibited significant anti-inflammatory effects in normal rats. The $3^{rd}$ experiment was found that the tested doses of the extract in diabetic rats induced a significant decrease in serum glucose, AST, ALT, triglycerides, cholesterol and LDL, while HDL caused a significant increase.

Effects of Magnesium Deficiency on Induction of Activity of Antioxidative Enzymes (Magnesium 결핍이 항산화효소의 활성유도에 미치는 영향)

  • Kim, B.C.;Lee, J.J.
    • Korean Journal of Weed Science
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    • v.17 no.4
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    • pp.431-438
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    • 1997
  • The influence of deficient(7${\mu}M$) and suffcient(1000${\mu}M$) magnesium(Mg) supply on the content of mg, chlorophyll, protein and the activity of superoxide dismutase(SOD), scavengers of superoxide radical($O_2$), and ascorbate peroxidase(AP), $H_2O_2$scavenging enzyme, and glutathione redutase(GR) were studied in pumpkin(Cudurbita moscata $D_{UCHESNE}$) plants over a 11-day period. Over the 11 days Period of growth in nutrient solution with sufficient and deficient Mg supply, the contents of Mg, chlorophyll protein and the activities of the antioxidative enzymes remained more or less constant in Mg-sufficient leaves. In Mg-deficient leaves, the contents of Mg, chlorophyll and protein was seriously decreased with time, however the activities of SOD, AP and GR highly enhanced compared to those of Mg-sufficient. The results indicated the stimulative effect of Mg deficiency on toxic oxygen species and scavenging enzymes in plants.

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Isolation and Cultural Characterization of Antibacterial Substance Producing Microbes (항균성 물질 생산 균주의 분리 및 배양학적 특성)

  • Park, Seok-Kyu;Cho, Young-Su;Shon, Mi-Yae;Gal, Sang-Wan;Lee, Sang-Won
    • Food Science and Preservation
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    • v.14 no.2
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    • pp.194-200
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    • 2007
  • In order to enhance the functionality and storage period of traditional fermented foods, the strain CH-14, which To enhance the quality of traditional fermented foods, and to lengthen acceptable storage periods, a bacterial strain, CH-14, showing potent enzyme activities and antibacterial capabilities, was isolated and characterize4 The bacterium wn Gram-positive, catalase-positive, oxidase-negative, formed endospores, expressed flagella, was rod-shaped, and had dimensions of 0.5 0.7m and 3.5 4.2m. The bacterium CH-14 was identified as Bacillus subtilis using Bergey's Manual of Systematic Bacteriology, Bergey's Manual of Determinative Bacteriology, and an API 50 CHL Carbohydrate Test Kit. An optimum growth medium contained 2% (w/v) cellobiose as a carbon source, a mixture of 0.5% (w/v) yeast extract and 0.5% (w/v) peptone as nitrogen sources, and 0.05% (w/v) $MgSO_4{\cdot}7H_2O$. The optimal culture temperature and the optimal initial pH were in the ranges of 30 $45^{\circ}C$ and 4.5 10.0, respectively. Maximum production of the antibacterial substance occurred after 24h of culture. The minimum inhibitory concentrations of the antibacterial substance were 5mg bacterial dry weight/mL against E. coli and P. mirabilis, and 10 mg/mL against S. aureus, S. enteritidis and V. parahaemolyticus.

The Effects of Feeding Feed Additives Containing Microorganisms on Characteristics of Excreta in Growing Pigs (육성돈에 미생물제제 급여시 분뇨 특성에 미치는 효과 연구)

  • Kwag, J.H.;Choi, D.Y.;Park, Ch.H.;Kim, J.H.;Jeong, K.H.;Yang, Ch.B.;Yoo, Y.H.;La, C.S.
    • Journal of Animal Environmental Science
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    • v.13 no.1
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    • pp.35-44
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    • 2007
  • The effects of microbial feedstuff additives on feed conversion rate and physical and chemical characteristics of excreta in growing pigs were investigated. Three different products (A, B and C) were compared. Microbial population tests showed B contained higher numbers of total bacteria, Lactobacillus spp. and yeasts. The amylase activity of B was also higher than that of A and C. The daily feed intake rates fer control, A, B and C were 2.06, 2.13, 2.17 and 2.34 kg, respectively. Pigs feed product C had the highest liveweight gain(2.89 kg). However, the results of feed conversion rate were not significantly different between treatments. Amount of faces excreted for control, A, B and C was 1.18, 1,19, 1.23 and 1.32 kg, respectively. Urine volume for control, A, B, and C was 1.91, 1.80, 2.19 and 2.31 kg respectively. Moisture content, T-N, $P_2O_5$ and $K_2O$ in pig manure were not significantly different between treatments. The range of BOD values was 63,453 to $73,758mg/\ell$ for faeces, and 5,678 to $7,428mg/\ell$, for urine. SS values of solid and liquid excreta ranged from 142,200 to 176,000 and from 710 to $1,025mg/\ell$, respectively.

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