• Title/Summary/Keyword: $Ni_2L_2H_{-3}^+$

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Molecular cloning, purification, expression, and characterization of β-1, 4-endoglucanase gene (Cel5A) from Eubacterium cellulosolvens sp. isolated from Holstein steers' rumen

  • Park, Tansol;Seo, Seongwon;Shin, Teaksoon;Cho, Byung-Wook;Cho, Seongkeun;Kim, Byeongwoo;Lee, Seyoung;Ha, Jong K.;Seo, Jakyeom
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.4
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    • pp.607-615
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    • 2018
  • Objective: This study was conducted to isolate the cellulolytic microorganism from the rumen of Holstein steers and characterize endoglucanase gene (Cel5A) from the isolated microorganism. Methods: To isolate anaerobic microbes having endoglucanase, rumen fluid was obtained from Holstein steers fed roughage diet. The isolated anaerobic bacteria had 98% similarity with Eubacterium cellulosolvens (E. cellulosolvens) Ce2 (Accession number: AB163733). The Cel5A from isolated E. cellulolsovens sp. was cloned using the published genome sequence and expressed through the Escherichia coli BL21. Results: The maximum activity of recombinant Cel5A (rCel5A) was observed at $50^{\circ}C$ and pH 4.0. The enzyme was constant at the temperature range of $20^{\circ}C$ to $40^{\circ}C$ but also, at the pH range of 3 to 9. The metal ions including $Ca^{2+}$, $K^+$, $Ni^{2+}$,$Mg^{2+}$, and $Fe^{2+}$ increased the endoglucanase activity but the addition of $Mn^{2+}$, $Cu^{2+}$, and $Zn^{2+}$ decreased. The Km and Vmax value of rCel5A were 14.05 mg/mL and $45.66{\mu}mol/min/mg$. Turnover number, Kcat and catalytic efficiency, Kcat/Km values of rCel5A was $96.69(s^{-1})$ and 6.88 (mL/mg/s), respectively. Conclusion: Our results indicated that rCel5A of E. cellulosolvens isolated from Holstein steers had a broad pH range with high stability under various conditions, which might be one of the beneficial characteristics of this enzyme for possible industrial application.

Optimized Design of Low Voltage High Current Ferrite Planar Inductor for 10 MHz On-chip Power Module

  • Bae, Seok;Hong, Yang-Ki;Lee, Jae-Jin;Abo, Gavin;Jalli, Jeevan;Lyle, Andrew;Han, Hong-Mei;Donohoe, Gregory W.
    • Journal of Magnetics
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    • v.13 no.2
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    • pp.37-42
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    • 2008
  • In this paper, design parameters of high Q (> 50), high current inductor for on-chip power module were optimized by 4 Xs 3 Ys DOE (Design of Experiment). Coil spacing, coil thickness, ferrite thickness, and permeability were assigned to Xs, and inductance (L) and Q factor at 10 MHz, and resonance frequency ($f_r$) were determined Ys. Effects of each X on the Ys were demonstrated and explained using known inductor theory. Multiple response optimizations were accomplished by three derived regression equations on the Ys. As a result, L of 125 nH, Q factor of 197.5, and $f_r$ of 316.3 MHz were obtained with coil space of $127\;{\mu}m$, Cu thickness of $67.8\;{\mu}m$, ferrite thickness of $130.3\;{\mu}m$, and permeability 156.5. Loss tan ${\delta}=0$ was assumed for the estimation. Accordingly, Q factor of about 60 is expected at tan ${\delta}=0.02$.

The Proteinase Distributed in the Intestinal Organs of Fish 3. Purification and Some Enzymatic Properties of the Alkaline Proteinases from the Pyloric Caeca of Skipjack, Katsuwonus vagans

  • PYEUN Jae-Hyeung;KIM Hyeung-Rak;HEU Min-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.21 no.2
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    • pp.85-96
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    • 1988
  • Purification and some properties of alkaline proteinases in the pyloric caeca of skipjack, Katsuwonus vagans, were investigated. Four alkaline proteinases, temporarily designated proteinases I, II, III and IV, were identified from the tissue extract of the pyloric caeca by ammonium sulfate fractionation, DEAE-Sephadex A-50 chromatography, and Sephadex G-100 and G-200 gel filtration. Result of disc-polyacrylamide gel electrophoretic analysis showed that the purified proteinases II and III were homogenous with the yields of $1.5\%\;and\;1.2\%$, and those specific activities were increased to 33 to 37 fold over that of the crude enzyme solution, respectively. Molecular weight of the proteinases II and III determined by sephadex G-100 gel filtration were 28,500 and 24,200, respectively. The optimum conditions for the caseinolytic activity of the two enzymes were pH 9.6 and $48^{\circ}C$. The reaction rates of the two alkaline proteinases were constant to the reaction time to 80 min in the reaction mixture of $3.4{\mu}g/ml$ of enzyme concentration and $2\%$ casein solution. The Km values against casein substrate determined by the method of Lineweaver-Burk were $0.56\%$ for proteinase II and $0.30\%$ for proteinase II. The proteinases II and III were inactivated under the presence of $Ag^+,\;Hg^{2+},\;Ni{2+},\;Fe^{2+},\;and\;Cu^{2+}$, and but activated by $Mn^{2+}\;and\;Ca^{2+}$ and markedly inhibited by the soybean trypsin inhibitor and N-p-toluenesulfonyl-L-lysine chloromethyl ketone. Therefore, the proteinases II and III were found to be a group of serine proteases and assured to be trypsin-like proteinases.

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Power Generating Characteristics of Anode-Supported SOFC fabricated by Uni-Axial Pressing and Screen Printing (일축가압/스크린인쇄 공정에 의해 제조된 음극지지형 SOFC의 출력특성)

  • 정화영;노태욱;김주선;이해원;고행진;이기춘;이종호
    • Journal of the Korean Ceramic Society
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    • v.41 no.6
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    • pp.456-463
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    • 2004
  • To enhance the performance of anode-supported SOFC, single cell fabrication procedure was changed for better and resulting power generating characteristics of single cell were investigated. Liquid condensation process was employed for the granulation of NiO/YSZ powder mixture and the produced powder granules were compacted into anode green substrate by uni-axial pressing. YSZ electrolyte was printed on green substrate via screen-printing method and co-fired at 1400$^{\circ}C$ for 3 h. LSM/YSZ composite cathode of which the composition and heat treatment condition was adjusted to minimize the polarization#resistance with AC-impedance spectroscopy, was screen printed. The final single cell size from this multi-step procedure was 5${\times}$5 $\textrm{cm}^2$ and 10${\times}$10 $\textrm{cm}^2$. The maximum power densities of 5${\times}$5 and 10${\times}$10 single cells were about 0.45 W/$\textrm{cm}^2$ and 0.22 W/$\textrm{cm}^2$ at 800$^{\circ}C$, which are two times superior than those from single cells fabricated by the conventional process in previous our work.

Characterization of Leaching Behaviour of Recycled Concrete for Environmental Assessment (용출특성규명을 통한 재생골재 환경성 평가)

  • Kang, S.H.;Lee, S.H.;Kwak, K.S.;Lee, J.Y.;Chung, M.K.
    • Journal of Korean Society of Environmental Engineers
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    • v.27 no.3
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    • pp.293-301
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    • 2005
  • We conducted several different leaching experiments for assessing the potential environmental risk when utilizing recycled concrete for stabilizing bridge pier. The methods include continuous batch leaching test (DIN 38414-S4), availability test (NEN 7341), pH-stat test (CEN/TC 292/WG6) and tank diffusion test (NEN 7345). The concentration ranges vary depending on the testing method. Nearly all the trace elements were low, some elements recording under detection limit. The maximum concentrations for trace elements leached throughout the whole tests are (as mg/L); Cd (0.029), Cu (0.437), Pb (0.14), Ni, Zn (0.95), Hg (0.005). Although the testing methods we used in this study are much more rigorous than other commonly adapted method including TCLP and domestic testing method for solid waste, the trace elemental concentrations are under the criteria for hazardous material set by the TCLP and domestic method. The result seems to suggest that applying the recycled concrete on stream water will be accepatable practice as for as trace elements are concerned. However, the influence of inorganics such as Ca, Mg, Ni and $SO_4^{2-}$ on aquatic ecology should be further examined.

Break Point Chlorination (BPC) Characteristics for Heavy Metals Removal in Plating Wastewater Treatment (염소산화공정을 이용한 도금폐수의 중금속 제거 특성)

  • Jung, Byung-Gil;Lee, Seung-Won;Yun, Kwon-Gam;Jung, Jin-Hee;Kim, Jeong-Woong;Choi, Young-Ik
    • Journal of Environmental Science International
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    • v.29 no.11
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    • pp.1055-1064
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    • 2020
  • In this research, heavy metals and T-P removal characteristics of plated wastewater are derived using BPC(Break Point Chlorination) process. AA sedimentation pond outflow(Influence) was evaluated for the removal efficiency of heavy metal(Ni) and T-P at a reaction time of 25 minutes by NaOCl input volume(9, 11, 13 and 15 mL). In the case, the higher the NaOCl input volumes, the higher the ORP values were maintained and the higher the removal efficiency tended to be. On the other hand, T-P was judged to have a low relationship between the ORP value and the removal efficiency. In addition, the efficiency of removal heavy metals and T-P in the plated wastewater by injecting 10 mL, 15 mL, 20 mL and 25 mL NaOCl, increased as the amount of NaOCl injected increased, the amount of NaOH input for pH increased. It was found that suspended solid in effluence also increased. It was also observed that the color of the plating wastewater changed from yellowish green to green to charcoal gray to black as the amount of NaOCl injected increased. Treatment characteristics of the reaction time, the longer the reaction time with the substance to be treated after the input of NaOCl, the more the heavy metal removal efficiency tended to increase. Through XRF analysis of the sludge, the constituents in the sludge such as NaCNO, CNCl, Na3PO4, CrO4, 2Na2CrO4 and 2NaNO3 will be analyzed in detail, and the mechanisms of the reaction between the plated wastewater and the complex compound will be elucidated.

Preparation and Characterization of Canned Kwamaegi(I) - Preparation and Characterization of Canned Seasoned Kwamaegi - (과메기 통조림의 제조 및 특성(I) - 조미과메기 통조림의 제조 및 특성을 중심으로 -)

  • Yoon, Ho-Dong;Shim, Kil-Bo;Noh, Yu-Ni;Kong, Cheung-Sik;Nam, Dong-Bae;Park, Tae-Ho;Kim, Jeong-Gyun
    • Journal of Fisheries and Marine Sciences Education
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    • v.23 no.4
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    • pp.662-672
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    • 2011
  • This study was investigated to obtain basic data which can be applied to processing of canned seasoned kwamaegi. Commercial Kwamaegi was cut into $2{\times}3cm$ lengths, filled 90 g into can (301-3) and added with 60 g water before precooking for 10 min. at $100^{\circ}C$. The precooked Kwamaegi was packed into the can, and added with 60 g seasoning sauces, which was prepared by mixing soy sauce 23%, monosodium glutamate (MSG) 2%, sorbitol 2%, sesame oil 1%, vinegar 2%, starch syrup 17%, sake 5%, water 48%. The cans were sealed using a vacuum seamer and then sterilized for various Fo values (Fo 8~12 min.) in a steam system retort at $121^{\circ}C$. The factors such as pH, VBN, amino-N, total amino acid, free amino acid, color value (L, a, b), texture profile, TBA value, mineral, sensory evaluation and viable bacterial count of the canned seasoned Kwamaegi produced with various sterilization condition(Fo 8~12 min.) were measured. There was no remarkable difference between sterilization conditions and sensual characteristics. The results showed that the product sterilized at Fo 8 min. was the most desirable because this condition is most economical.

Solid-Phase Refolding of Poly-Lysine fusion Protein of hEGF and Angiogenin (Poly-lysine이 연결된 hEGF와 angiogenin의 융합단백질의 고체상 재접힘)

  • Park, Sang-Joong;Ryu, Kang;Suh, Chang-Woo;Chai, Young-Gyu;Kwon, Oh-Byung;Park, Seung-Kook;Lee, Eun-Kyu
    • KSBB Journal
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    • v.17 no.2
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    • pp.153-157
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    • 2002
  • A fusion protein, consisting of a human epidermal growth factor as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as an inclusion body in recombinant E. coli, yet when the conventional solution-phase refolding process was used the refolding yield was very low due to severe aggregation, probably because of the opposite surface charge resulting from the vastly different pl values of each domain. Accordingly the solid-phase refolding process, which exploits the ionic interactions between a solid matrix and the protein, was tried, however the ionic binding yield was also very low regardless of the resins and pH conditions used. Therefore, to provide a higher affinity toward the solid matrix, six Iysine residues were tagged to the N-terminus of the hEGF domain. When cation exchange resins, such as heparin- or CM-Sepharose, were used as the matrix, the adsorption capacity increased 2.5~3-fold and the subsequent refolding yield increased nearly 15-fold compared to the conventional process. A similat result was also obtained when an Ni-NTA metal affinity resin was used.

Liquid-Composting Conditions of By-product Obtained from Degradation of Animal Carcass for Agriculture Recycling (폐가축사체의 농업적 재활용을 위한 가축사체 액상부산물의 액비화 조건 구명)

  • Seo, Young-Jin;Seo, Dong-Cheol;Kang, Se-Won;Lee, Sang-Gyu;Park, Ju-Wang;Choi, Ik-Won;Sung, Hwan-Hoo;Kang, Seog-Jin;Cho, Ju-Sik
    • Korean Journal of Environmental Agriculture
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    • v.32 no.4
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    • pp.348-354
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    • 2013
  • BACKGROUND: Globally, concern about emerging infectious diseases of livestock is growing. For the disposal of the animal carcass, it is necessary to recycle the carcass into an agriculturally usable product. The objective of this study was to investigate the composting conditions of liquid by-product obtained from degradation of animal carcass. METHODS AND RESULTS: Optimum conditions of liquid fertilizer were investigated using different microorganisms, pHs, and volumes of microorganisms (Lactobacillus rhamnosus+Pichia deserticola). Based on the results from the optimum conditions, compost maturity and quality of liquid fertilizer were evaluated for 112 days. The compost maturity of liquid fertilizer were higher in the order of LP(Lactobacillus rhamnosus + Pichia deserticola) > BC(Bacillus cereus) > BS(Bacillus subtilis). The optimum condition under different volumes of LP was injection of 0.5 mL/100 mL. The compost maturity under different pHs were higher in the order of pH 7 > $$5{\geq_-}9{\frac{._-}{.}}11$$. The liquid by-product at 56 days after composting was completely decomposed. The concentrations of T-N, T-P and $K_2O$ in liquid fertilizer at 56 days were 0.94, 0.17 and 3.78%, respectively, and the sum of those concentrations was 4.89%. CONCLUSION(S): Liquid fertilizer of by-product using pig carcass was decomposed with optimum conditions(LP, pH 7, injection of 0.5 mL/100 mL) in 56 days after composting, and was suitable for official standard of commercial fertilizer.

Studies on Antimutagenic Effects and Gene Repair of Enzymatic Browning Reaction Products (효소적 갈변반응 생성물의 돌연변이 억제효과 및 유전자 수복에 관한 연구)

  • Ham, Seung-Shi;Kim, Sung-Wan;Kim, Young-Myung
    • Korean Journal of Food Science and Technology
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    • v.22 no.6
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    • pp.632-639
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    • 1990
  • The biological activities of twelve different kinds of enzymatic browning reaction products(EBRP), which resulted from the reactants four kinds of polyphenols with polyphenol oxidase extracted from Ligularia fischeri, pimpinella brachycarpa and Aster scaber of edible mountain herbs. All of twelve samples did not show any mutagenic effect in the spore rec-assay, Ames mutagenicity test and DNA breaking test. However metal ions such as $Cu^{2+},\;Fe^{2+}$, and $Ni^{2+}$ were increased the DNA breakage in rec-assay. The EBRPs inhibited the mutagenicities induced by $benzo({\alpha})pyrene (B({\alpha})P)$, 3-amino-1,4-dimethyl-5H-pyrido-[4,3-b]indole(Trp-P-1) and 2-aminofluorene(2-AF) in Salmonella/microsome assay system with S-9 mix. In effects of EBRPs on the DNA repair system, the activity of EcoRI was highly inhibited and that of $T_{4}$ DNA ligase was inactivated by addition of EBRPs. The results of transformation ratio of plasmid pGA658 into E. coli HB 101 was significantly decreased by the reaction products of S. brachycarpa polyphenoloxidase (PPO). When UV light was exposed to the mixture of DNA and EBRP before the thanformation, the reaction products from L. fischeri PPO with pyrogallol, catechol and hydroxyhydroquinone stimulated transformation ratio.

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