• Title/Summary/Keyword: bio-ion

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Purification, and Biochemical and Biophysical Characterization of Cellobiohydrolase I from Trichoderma harzianum IOC 3844

  • Colussi, Francieli;Serpa, Viviane;Da Silva Delabona, Priscila;Manzine, Livia Regina;Voltatodio, Maria Luiza;Alves, Renata;Mello, Bruno Luan;Nei, Pereira Jr.;Farinas, Cristiane Sanches;Golubev, Alexander M.;Santos, Maria Auxiliadora Morim;Polikarpov, Igor
    • Journal of Microbiology and Biotechnology
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    • v.21 no.8
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    • pp.808-817
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    • 2011
  • Because of its elevated cellulolytic activity, the filamentous fungus Trichoderma harzianum has a considerable potential in biomass hydrolysis applications. Trichoderma harzianum cellobiohydrolase I (ThCBHI), an exoglucanase, is an important enzyme in the process of cellulose degradation. Here, we report an easy single-step ion-exchange chromatographic method for purification of ThCBHI and its initial biophysical and biochemical characterization. The ThCBHI produced by induction with microcrystalline cellulose under submerged fermentation was purified on DEAE-Sephadex A-50 media and its identity was confirmed by mass spectrometry. The ThCBHI biochemical characterization showed that the protein has a molecular mass of 66 kDa and pI of 5.23. As confirmed by smallangle X-ray scattering (SAXS), both full-length ThCBHI and its catalytic core domain (CCD) obtained by digestion with papain are monomeric in solution. Secondary structure analysis of ThCBHI by circular dichroism revealed ${\alpha}$- helices and ${\beta}$-strands contents in the 28% and 38% range, respectively. The intrinsic fluorescence emission maximum of 337 nm was accounted for as different degrees of exposure of ThCBHI tryptophan residues to water. Moreover, ThCBHI displayed maximum activity at pH 5.0 and temperature of $50^{\circ}C$ with specific activities against Avicel and p-nitrophenyl-${\beta}$-D-cellobioside of 1.25 U/mg and 1.53 U/mg, respectively.

Establishment of Analytical Method for Pencycuron in Representative Agricultural Commodities by High-Performance Liquid Chromatography (대표 농산물 중 살균제 Pencycuron의 HPLC 정밀 잔류분석법 개발)

  • Lee, Hyeri;Choi, Hoon;Kim, Byung-Joon;Kim, Eunhye;Kim, Su-Hee;Lee, Jin-Beom;Lee, Young Deuk;Kim, Jeong-Han
    • The Korean Journal of Pesticide Science
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    • v.21 no.1
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    • pp.75-83
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    • 2017
  • The single residue analytical method was developed for determining fungicide pencycuron residues in various agricultural commodities with high-performance liquid chromatography (HPLC). Pencycuron residue was extracted with acetone from representative crops such as Korean cabbage, apple, brown rice and green pepper. After ethyl acetate/n-hexane partition and subsequent clean-up with silica gel chromatography, pencycuron residue was quantified by reversed phase HPLC with UV detection at 240 nm. The suspected residue of pencycuron was confirmed using selected-ion monitoring (SIM) LC/mass spectrometry (MS). Instrumental limit of quantitation (ILOQ) and method LOQ (MLOQ) were set at 2 ng and 0.02 mg/kg, respectively. Overall recoveries of pencycuron from different crop samples fortified at three levels (MLOQ, 10MLOQ, 100MLOQ) were 72~108%. This proposed method could be useful as official analytical method for quantification of pencycuron residues in agricultural commodities.

A Study on the Separation of Electrolyte from Amino Acid Solution through Electrodialysis (전기투석법을 이용한 아미노산으로부터 전해질 분리정제에 관한 연구)

  • 김석곤;한정우;김한성;전경용;조영일
    • Membrane Journal
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    • v.4 no.3
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    • pp.163-170
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    • 1994
  • The separation of inorganic salt from amino acid solution using was performed electrodialysis. In order to review the availability of electrodialysis using isoelectric point of amino acid as a bio-separation technique, electrodialysis stacks were designed using ion exchange membrane. Separation of NaCl from amino acid solution was performed in the condition similar to amino acid fermentation process. To obtain otimum conditions of separation, leakage of amino acid depending of pH and limiting current density were measured. On the basis of optimum condition, removal of NaCl and leakage of amino acid were investigated quantitatively in batch and continuous process, and current efficiencies were also obtained. As a result of batch experiment for 11 hours each amino acid solution, removal efficiencies of NaCl were in the ranges of 96.1~96.2%. Amino acid leakage rate of glycine, methionine, alanine were 2.5, 1.7, 2.0% respectively. Current efficiencies were in the ranges of 44.5~44.6%. As a result of continuous experiment in various flow rate of each amino acid solution, it took 120 ~ 150 min to reach to steady state. Removal efficiency of NaCl was increased as the flow rate was decreased, but current efficiency was decreased. At the steady states, there were no leakage of amino acid.

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Removal Characteristics of Dichloroacetic Acid at Different Catalyst Media with Advanced Oxidation Process Using Ozone/Catalyst (담지체를 달리한 오존/촉매 AOP공정에서 디클로로아세트산의 제거 특성)

  • Park, Jin Do;Lee, Hak Sung
    • Applied Chemistry for Engineering
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    • v.20 no.1
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    • pp.87-93
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    • 2009
  • Pd/activated carbon (Pd/AC) and Pd/alumina (Pd/AO) catalysts were prepared by the impregnation of palladium into activated carbon and alumina. The catalytic characteristics according to the kinds of support materials were compared. The decomposition efficiencies of ozone according to kinds of support materials are about the same when these were compared by adding 10 g of catalysts into the water saturated with ozone. The decomposition efficiencies and the oxidation characteristics (TOC, $COD_{Cr}$) of dichloroacetic acid were compared with the ozone only process and the catalytic ozonations using Pd/activated carbon and Pd/alumina catalysts. The decomposition efficiencies of dichloroacetic acid by catalytic ozonations were better than the one by ozone only process, but there was slight difference of the one between Pd/activated carbon and Pd/alumina catalyst. The decomposition efficiency of dichloroacetic acid was increased with increasing ozone dose at a constant concentration of dichloroacetic acid, but the one was little increased with increasing ozone dose at more than 1.0 L/min of ozone dose. It was seemed that the bicarbonate and the chloric ion formed throughout the decomposition of dichloroacetic acid acted as the scavenger of hydroxyl radical.

The Leaching of Valuable Metal from Mine Waste Rock by the Adaptation Effect and the Direct Oxidation with Indigenous Bacteria (토착박테리아의 중금속 적응효과와 직접산화작용에 의한 폐광석으로부터 유용금속 용출)

  • Kim, Bong-Ju;Cho, Kang-Hee;Choi, Nag-Choul;Park, Cheon-Young
    • Journal of the Mineralogical Society of Korea
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    • v.28 no.3
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    • pp.209-220
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    • 2015
  • The aim of this study was leaching valuable metal ions from mine waste rocks which were abandoned mine site using indigenous aerobic bacteria. In order to tolerate the the indigenous aerobic bacteria to the heavy metal ions they were repeatedly adapted in $CuSO_4{\cdot}5H_2O$ environment. As the repeated generation-adaptation progressed, the pH values of the growth-medium were gradually decreased. During bio-leaching experiments with indigenous aerobic bacteria raised in a heavy metal ion environment for 42 days, the pH of the leaching solution was decreased while increasing the adaptation period. The indigeous bacteria were much more active on the surface of Younhwa waste rocks which contained relatively few the chalcopyrite and Cu content than the Goseong mine waste rocks, and also the amount of Cu and Fe ions were leached more in the Younhwa sample(leaching rate of 92.79% and 55.88%, respectively) than the Goseong sample(leaching rate of 66.77% and 21.83%, respectively). Accordingly, it is confirmed that valuable metal ions can be leached from the mine waste rocks, if any indigenous bacteria which inhabits a mine environment site for a long time with heavy metal ions can be used, and these bacteria can be progressively adapted in the growth-solutions containing the target heavy metals.

Purification and Characterization of β-Lactamase Secreted from Bacillus sp. J105 Strain having β-Lectam Antibiotics Resistance. ((β-lactam계 항생물질 저항성을 지닌 Bacillus sp. J105 균주로부터 분비되는 베타 락탐 분해효소의 정제 및 특성)

  • Cho, Kyeong-Soon;Kang, Byoung-Won;Seo, Min-Jeong;Lee, Young-Choon;Lee, Jai-Heon;Joo, Woo-Hong;Choi, Yung-Hyun;Lim, Hak-Seob;Kim, Jeong-In;Seo, Kwon-Il;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.18 no.6
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    • pp.845-851
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    • 2008
  • ${\beta}-Lactamase$, secreted from Bacillus sp. J105 strain was purified to a single band on SDS-PAGE by ammonium sulfate precipitation, ion exchange column chromatography and gel-filtration. The molecular weight of the purified enzyme was 31 kDa on SDS-PAGE and its isoelectric point was 7.35. Optimal pH and temperature for enzymatic reaction were 5 and $40^{\circ}C$, respectively. As a result of total amino acid composition analysis of the purified enzyme, Gly and Ala were occupied 14.1 and 13.3 mole %, respectively. Km and Vmax value of purified enzyme were 1.33 mM and 0.36 mM/ml using ampicillin as a substrate, respectively.

Phytochemical-based Tannic Acid Derivatives as Draw Solutes for Forward Osmosis Process (정삼투 공정의 유도용질로서의 식물 화학물질 기반의 탄닌산 유도체)

  • Kim, Taehyung;Ju, Changha;Kang, Hyo
    • Membrane Journal
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    • v.28 no.3
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    • pp.157-168
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    • 2018
  • Potassium tannate (TA-K), which is prepared by base treatment of the bio-renewable tannic acid (TA), was evaluated for its potential application as a draw solute for water purification by forward osmosis. The forward osmosis and recovery properties of TA-K were systematically investigated. In the application of forward osmosis through the active layer facing feed solution (AL-FS) method, the water flux of TA-K draw solution was significantly higher than that of the TA draw solution, while that of the latter was not identified. At a low concentration of 100 mM, the osmotic pressure (1,135 mOsmol/kg) of the TA-K draw solution was approximately 6.5 times that (173 mOsmol/kg) of the NaCl draw solution. Furthermore, the water flux and specific salt flux (6.14 LMH, 1.26 g/L) of the TA-K draw solution at 100 mM were approximately 2.5 and 0.5 times those of the NaCl draw solution (2.46 LMH, 2.63 g/L) at the same concentration, respectively. For reuse, TA-K was precipitated by using a metal ion and recovered through membrane filtration. This study demonstrates the applicability of a phytochemical material as a draw solute for forward osmosis.

Characterization of a Fibrinolytic Enzyme from Pickled Anchovy (멸치젓갈유래의 혈전용해호소에 대한 특성)

  • Yang Woong-Suk;Lim Hak-Seob;Chung Kyung Tae;Kim Young-Hee;Huh Man Kyu;Choi Byung Tae;Choi Yung Hyun;Jeong Yong Kee
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.434-438
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    • 2005
  • In the previous study, we isolated a myulchikinase (MK), which has fibrinolytic activity and cytotoxicity to the tumor cell line, from myl- chi-jeot-gal. In this study, the effect of NaCl concentration, metallic ions, pH, temperature, and plasminogen on the activity of MK was analysed. The MK activity was maintained at least $80\%$ activity up to $30\%$ NaCl, which indicates that the enzyme may be halotolerant. The optimal pH and temperature were 8 and $40^{\circ}C$, respectively. The fibrinolytic activity of MK was completely inhibited with 0.5 mM $Hg^{2+}$ and inhibited to $50^{\circ}C$ with 1 mM $Cu^{2+}\;and\;Zn^{2+}$. The MK showed strong activity in plasminogen- rich fibrin plate but not in plasminogen-free fibrin plate. The result indicates that the MK may be a plasminogen activator type fibrinolytic enzyme.

DNA Breakage by Salvianolic acid B in the Presence of Cu (II) (구리이온(II)이 존재할 때 Salvianolic acid B에 의한 DNA 절단)

  • Lee, Pyeongjae;Moon, Cheol;Choi, Yoon Seon;Son, Hyun Kyu
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.2
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    • pp.205-210
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    • 2018
  • Salvianolic acid B, which is a compound in the Salvia miltiorrhiza, has diverse biological activities, In particular, the antioxidative effects were reported to be involved in the protection of hepatocytes, neurons, and various cell types. On the other hand, some phenolic compounds, such as ferulic acid, which is regarded as an antioxidant, plays a pro-oxidative role in the specific transitional metal environment, which could explain the anticancer effect. This study examined the pro-oxidative effects of salvianolic acid B in the presence of $Cu^{2+}$. Treatment with both salvianolic acid B and $Cu^{2+}$ induced the transition of supercoiled DNA to the open circular or linear form but not in the sole salvianolic acid B or $Cu^{2+}$ treatments. Salvianolic acid B reduced the $Cu^{2+}$ to $Cu^+$ using neocuproine, a $Cu^+$ specific chelator. In addition, catalase, an enzyme that breaks down the $H_2O_2$ to water and molecular oxygen, inhibited the DNA breakage. $H_2O_2$, a reactive oxygen species, has detrimental effects on biological molecules, particularly DNA. Overall, the reduction of $Cu^{2+}$ by salvianolic acid B could lead to the production of $H_2O_2$ followed by DNA breakage. These results suggest that the pro-oxidative effects could be the one of the anti-cancer mechanisms of salvianolic acid B, which remains to be explained.

Quality characteristics and antioxidant capacities of oriental melon wine depending on pretreatments (전처리 방법에 따른 참외와인의 품질특성과 항산화능)

  • Hwang, Hee-Young;Ha, Hyoung-Tae;Ha, Se-Bi;Seong, Gi-Un;Hwang, In-Wook;Chung, Shin-Kyo
    • Food Science and Preservation
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    • v.22 no.3
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    • pp.421-427
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    • 2015
  • Oriental melon (Cucumis melo L. var. makuwa) has been widely consumed as various processed foods, such as dried products, jelly, wine, juice, and vinegar, in Asian countries. In fruit processing, blanching and pressure treatments affect its quality, such as antioxidant activities, sensory characteristics, and etc. This study was conducted to evaluate the effects of blanching and pressure pre-treatments on oriental melon wine-making (BP, blanching and pressure filtration; BNP, blanching and non-pressure filtration; NBP, non-blanching and pressure filtration; and NBNP, non-blanching and non-pressure filtration). Physicochemical properties and antioxidant capacities by ${\alpha}$, ${\alpha}$-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, ferric ion reducing antioxidant power (FRAP) assay, and total phenolic and total flavonoid contents were measured for comparison of the different pre-treatment methods. After the aging process, the alcohol contents and pH values showed no statistical differences, whereas the amount of soluble solids, reducing sugar, and titratable acidity were slightly different among the pre-treatments (p<0.05). The samples with blanching pre-treatment showed higher antioxidant capacities than those of other pre-treatments. In the sensory evaluation, the BNP was the best in overall acceptability. Thus, this study showed that the blanching treatment enhanced the antioxidant capacities and sensory qualities of the oriental melon wine.