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Purification and Characterization of Exo-xylanase from Escherichia coli Cells Harboring the Recombinant Plasmid pMGl (재조합 균주 Escherichia coli가 생산하는 Bacillus stearothermophilus Exo-xylanase의 정제 및 특성)

  • 문애란;최용진
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.574-582
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    • 1992
  • Exo-xylanase encoded by the xylA gene of Bacillus stearothermoPhillus was produced from Escherichia coli ]M109 carrying a recombinant plasmid pMGL Synthesis of the enzyme was observed to be cell-associated, and about 94% of the enzyme synthesized was located in the cytoplasmic region. The maximum production was attained when the E. coli strain was grown at $37^{\circ}C$ for 8 hours on the medium containing 0.5% fructose, 1.0% tryptone, 1.0% sodium chloride, and 0.5% yeast extract. The exo-xylanase was purified to homogeneity using a combination of salting out with ammonium sulfate, DEAE-Sepharose CL-6B ion exchange chromatography, Sephadex G-IOO gel filtration, and Sephadex G-150 gel filtration. The' purified enzyme was most active at pH 6.0 and $45^{\circ}C$. $Ca^{2+}$ and $Co^{2+}$ activated the exo-xylanase activity by about 20% while $Ag^{2+}$, $Fe^{2+}$, $Mg^{2+}$ and $Zn^{2+}$ inhibited the enzyme activity by up to 60%. The $K_m$, value on p-nitrophenyl-$\beta$-D-xylanopyranoside was 2.75 mM. The enzyme had a pI value of 4.7. The estimated molecular weight of the native protein was 200,000 daL SDS-polyacrylamide gel electrophoresis analysis suggested that the native enzyme was a trimer composed of three identical 66,000 da!. polypeptides. The purified enzyme efficiently converted all the xylo-oligosaccharides tested to xylose. It was also confirmed that the enzyme split xylans in an exo-manner even though the degree of hydrolysis was fairly low. The xylanolytic enzyme was, therefore, classified to be one of the few bacterial exo-xylanases lacking transferase activity.

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Association between the Polymorphism of the Fatty acid binding protein 5 (FABP5) Gene within the BTA 14 QTL Region and Carcass/Meat Quality Traits in Hanwoo (한우 14번 염색체 QTL 영역내 Fatty acid binding protein 5 유전자의 다형성과 도체 및 육질 형질과의 관련성 분석)

  • Heo, Kang-Nyeong;Kim, Nam-Kuk;Lee, Seung-Hwan;Kim, Nam-Young;Jeon, Jin-Tae;Park, Eung-Woo;Oh, Sung-Jong;Kim, Tae-Hun;Seong, Hwan-Hoo;Yoon, Du-Hak
    • Journal of Animal Science and Technology
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    • v.53 no.4
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    • pp.311-317
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    • 2011
  • The aim of this study was to evaluate the association between economic traits of Korean cattle (Hanwoo) and genetic variation in fatty acid binding protein 5 (FABP5) gene within QTL region of carcass weight and marbling score traits on BTA 14. We sequenced for detection of single nucleotide polymorphism (SNP) with 24 unrelated Hanwoo samples and identified four SNPs (-1141A>G, 949A>G, 969A>G and 1085C>G). Relationship between the genotypes of 583 Hanwoo individuals by PCR-RFLP and economic traits were analyzed by the mixed regression model implemented in the ASReml program. As the result of statistical analysis, SNP1 (-1141A>G) showed significant effect (p<0.003) on marbling score (MS) and SNP2 (949A>G) showed significant effect (p<0.034) on eye muscle area (EMA). Further studies are required to validate the significant SNPs in a bigger population, but the SNPs (-1141A>G and 949A>G) of FABP5 could be a genetic marker to estimate molecular breeding value (MEBV) for carcass traits in Hanwoo.

The Effect of Biodiesel Blend Fuels As Reductants on NOx Conversion Efficiency of HC_SCR (환원제로서 바이오디젤 혼합연료가 HC-SCR의 NOx 변환효율에 미치는 영향 연구)

  • Song, Hoyoung;Lee, Minho;Kim, Kiho
    • Journal of Energy Engineering
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    • v.24 no.4
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    • pp.140-145
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    • 2015
  • This study was aimed at analyzing NOx conversion characteristics in the HC-SCR with biodiesel content changes of the secondary fuel injection (BD0, BD10, BD25). Test conditions for temperature were set to $290^{\circ}C$, $320^{\circ}C$ and $350^{\circ}C$ considering the upstream temperature of a HC-SCR, distillation of the secondary injected fuels and etc. The amount of fuel injection was adjusted with a fixed space velocity of 55,000(1/h). According to the test results of distillation, the T90 was the same level about $350^{\circ}C$ on all test fuels and the amount of evaporation was reduced at lower than $350^{\circ}C$ temperature condition with increasing biodiesel content. As biodiesel content which is mixed with the secondary injected fuel is increased, NOx reduction efficiency was determined to decrease. The difference of the Nox reduction ratio in a high temperature condition($320^{\circ}C$ and $350^{\circ}C$) than the low temperature($290^{\circ}C$) was more significant. These results are thought to be poor evaporation properties (distillation) and high molecular weight of the biodiesel.

A Study on Properties of the Urethane Prepolymer Synthesis with Polyether-diol and Aromatic Diisocyanate System (폴리에테르-디올과 방향족 디이소시아네이트계의 우레탄 프리폴리머 합성에 따른 특성연구)

  • Lee, Hyun-Joo;Kim, Kwang
    • Applied Chemistry for Engineering
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    • v.9 no.4
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    • pp.491-496
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    • 1998
  • The composition of isocyanates and polyols influence prepolymeric properties of adhesive and calking sealant based on polyurethanes (PU). One component moisture curing prepolymers, which reacted with surface humidity of substrate, were synthesized in several kinds of composition. Reactivity, structural change and properties of the prepolymers were studied as a preliminary step to manufacture PU based adhesive and sealant. To study the effects of mole ratio ([NCO]/[OH]), we used toluene diisocyanate (TDI), 4, 4'-diphenylmethane diisocyanate (MDI), and ether-polyols such as PTMG and PPG which have good resistance to hydrolysis and excellent low-temperature properties. The each prepolymers could be prepared in different molecular weight without any significant structural change. The mole ratio 1.78 of [NCO] to [OH] showed the fastest reactivity. It was confirmed that effect of polyols was larger than that of isocyanates on the prepolymer in reactivity. Several kinds of compounds were manufactured with each prepolymer, and tensile and properties were tested. And the optimum quantity of curing accelerator for the PU was 0.05~0.1%. In the tensile test, TDI based PU was superior to MDI based PU, and also PTMG based PU was superior PPG based PU.

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Synthesis of Polyamine Type Flocculant and Properties in Potable Water Treatment (Polyamine계 고분자 응집제의 합성 및 상수 처리 특성)

  • Park, Lee-Soon;Shin, June-Ho;Choi, Sang-June;Shin, Myung-Chul;Lee, Seok-Hun
    • Applied Chemistry for Engineering
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    • v.9 no.4
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    • pp.542-547
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    • 1998
  • Polyamine type polymer flocculants were synthesized and their characterization, viscosity and flocculation studies were conducted. In order to increase the molecular weight of polyamine flocculant which was prepared by polycondensation reaction from dimethylamine and epichlorohydrin, a small amount of 1,6-hexanediamine was employed. The incorporation of 1,6-hexanediamine up to 5.5 mole % replacing corresponding part of dimethylamine gave a branched type polyamine sample with increased intrinsic viscosity ([${\eta}$]=0.46 in 1 wt % aqueous NaCl solution). The amount of 1,6-hexanediamine above 5.5 mole %, however, resulted in gelation during polymerization. Utilizing raw water from Maegok potable water treatment plant, it was found that the addition of polyamine flocculant at a concentration of 1 mg/L level could reduce the amount of polyaluminum chloride (PAC) inorganic flocculant by half (15 mg/L). It was also observed that the incoporation of polyamine flocculant at 1 mg/L level was effective in the higher pH raw water, while PAC inorganic flocculating agent alone was not effective.

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Syntheses and Properties of Copolymers of Tetramethyldisiloxane and 1,4-Bis(dimethylhydroxysilyl)benzene (Tetramethyldisiloxane-1,4-bis(dimethylhydroxysilyl)benzene 공중합체의 합성 및 특성)

  • Kim, Sun-ll;Yun, Young-Jae;Na, Jae-Woon
    • Applied Chemistry for Engineering
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    • v.7 no.3
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    • pp.490-495
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    • 1996
  • Low molecular weight linear chlorine terminated siloxanes (yields;71.2~86.5%) were prepared by reactions of cyclotri-, cyclotetra- and cyclopentasiloxane with dimethyldichlorosilane in the presence of pyridine N-oxide catalyst. The amine terminated siloxane oligomers were obtained in good yields(76.2~85.3%) by the reaction of linear chlorine terminated siloxanes with dimethylamine at $0^{\circ}C$. In this investigation, we have studied on the syntheses and properties of copolymers (yields;58.0~71.0%) obtained from the reaction of amine terminated siloxane oligomers with 1,4-bis(dimethylhydroxysilyl)benzene. The structures and properties of the copolymers were examined by FT-IR, $^1H$-NMR, TGA and DSC. Initial degradation temperatures($T_D{^i}$) of the polymer I and IV were confirmed as 476 and $485^{\circ}C$, respectively. The thermal stabilities of the polymers were found to be increased with increasing n of $(R_2SiO)_n$. The glass transition temperatures(Tg) of the polymers were increased with decreasing n of $(R_2SiO)_n$, and the lowest Tg revealed $-76^{\circ}C$ when n=5.

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Study on Meat Tenderness of a Pretense Extracted from Domestic Pear (국산배에서 추출한 단백질 분해효소의 식육 연화제로서의 활용에 관한 연구)

  • Han Seung K.;Chin Koo B.
    • Food Science of Animal Resources
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    • v.24 no.4
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    • pp.326-328
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    • 2004
  • Domestic pear has been reported that it contained a pretense, which used for tenderizer of meat, however no researches for optimum level of the enzyme with maximum tenderness effect have been studied. Thus, this study was peformed to determine the optimum level of a protease for meat tenderness. Moisture contents (%) of domestic pears was determined. A pretense was homogenized in a mixer and centrifuged at 10,000 G for 1hr. After taken the supernatant, dialysis was conducted to remove salts and sugars, and freeze-dried. Then, various level (0.05, 0.1 and 0.2%) of the purified pretense were added to pork loins (3cm thickness). Then, pork samples were boiled at 80 for 12 min in a water bath to reach the interval temperature of 71 and chilled in an ice. Moisture contents (%) of domestic pears ranged from 87.2 and 87.8%. No differences in cooking loss of pork meats were observed (p>0.05) among various levels of a pretense. After centrifugation, the protein concentrations of a protease showed from 5.96 $\mu\textrm{g}$/fmL to 7.25 $\mu\textrm{g}$/mL. Increased level of a pretense up to 0.1% reduced (p<0.05) the shear value (kg/g), however no further reduction of shear value was observed at the level of higher than 0.1% of the purified pretense. The approximate molecular weight of the pretense analysed by sodium-dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was 30 kDa. These results suggest that the optimum level of a pretense for the maximum effect of meat tenderness is above 0.1%. Further research will be peformed to determine the effect of various domestic pears and ingredients, such as salt and phosphate, on meat tenderness.

Electrochemical Characteristics of Lithium Battery Anode Materials Using Petroleum Pitches (석유계 피치를 사용한 리튬전지 음극소재의 전기화학적 특성)

  • Hwang, Jin Woong;Lee, Jong Dae
    • Applied Chemistry for Engineering
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    • v.28 no.5
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    • pp.534-538
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    • 2017
  • In this study, the molecular weight controlled pitches derived from pyrolyzed fuel oil (PFO) were prepared using solvent extraction and were carbonized. Electrochemical characteristics of lithium battery anode materials were investigated using these petroleum pitches. Three pitch samples prepared by the thermal reaction were 3903 (at $390^{\circ}C$ for 3 h), 4001 (at $400^{\circ}C$ for 1 h) and 4002 (at $400^{\circ}C$ for 2 h). The prepared hexane insoluble pitches were analysed by XRD, TGA, SEM and Gel permeation Chromatography (GPC). The electrochemical characteristics of the PFO-derived pitch as an anode material were investigated by constant current charge/discharge, cyclic voltammetry and electrochemical impedance tests. The coin cell using pitch (4001) and the electrolyte of $LiPF_6$ in organic solvents (EC : DMC = 1 : 1 vol%, VC 3 wt%) has better initial capacity (310 mAh/g) than that of other pitch coin cells. Also, this carbon anode showd a high initial efficiency of 82%, retention rate capability at 2 C/0.1 C of 90% and cycle retention of 85%. It was found that modified pitches improved the cycling and rate capacity performance.

Expression of Cu/Zn SOD according to H2O2 in Hepatoma cell line (Hepatoma 세포주에서 H2O2 처리에 의한 Cu/Zn SOD의 발현)

  • Kim, Young-Min;Seo, Won-Sook
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.230-234
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    • 2007
  • Oxygen is required for many important aerobic cellular reactions, it may undergo electrontransfer reactions, which generate highly reactive membrane-toxic intermediates (reactive oxygen species, ROS), such as hydrogen peroxide, singlet oxygen, superoxide radical, hydroxyl radical, hydroperoxyl radical, hydroxy ion. Various mechanisms are available to protect cells against damage caused by oxidative free radicals, including scavenging enzyme systems such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). This antioxidant defense system is a very complex and finely tuned system consisting of enzymes capable of detoxifying oxygen radicals as well as low molecular weight antioxidants. In addition, repair and turnover processes help to minimize subcellular damage resulting from free radical attack. $H_2O_2$,one of the major ROS, is produced at a high rate as a product of normal aerobic metabolism. The primary cellular enzymatic defense systems against $H_2O_2$ are the glutathione redox cycle and catalase. From Northern blot analysis of total RNAs from cultured cell with $H_2O_2$ treatment, various results were obtained. Expression of Cu/Zn SOD decreased when cell passage increased, but the level of the Cu/Zn SOD was scarcely expressed in 35 passage.

Effects of Chitosan on the Toxicity of Environmental Pollutants (해양바이오물질이 PCB의 독성작용에 미치는 영향)

  • Lee, Hyon-Gyo;Kim, Hae-Young;Yang, Jae-Ho
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.2
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    • pp.102-107
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    • 2007
  • Environmental contamination becomes a great public concern as our society gets industrialized rapidly. The present study examine the role of chitosan in a effort to intervene the environmental pollutant-induced toxicity. PCB-induced neurotoxicity with respect to the PKC signaling was examined. Since the developing neuron is particularly sensitive to PCB-induced neurotoxicity, we isolated cerebellar granule cells derived from 7-day old SD rats and grew cells in culture for additional 7 days to mimic PND-14 conditions. PCB showed the alteration of PKC signaling pathway. The alteration was structure-dependent. Mono-ortho-substituted congeners at a high dose showed a significant increase of total PKC activity at [$^3H$]PDBu binding assay, indicating that mono-ortho-substituted congeners are more neuroactive than non-ortho-substituted congeners in neuronal cells. PKC isoforms were immunoblotted with respective monoclonal antibodies. PKC-beta II and -epsilon were activated with mono-ortho-substituted congeners exposure. The result suggests that the position with ortho has a higher potential of altering the signaling pathway. Alteration of PKC was blocked with treatment of high molecular weight of chitosan. The study demonstrated that the ortho position in PCBs are important in assessing the structure-activity relationship. The results suggest a potential use of marine bioactive materials as a means of nutritional intervention to prevent the harmful effects of pollutant-derived toxicity.

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