• Title/Summary/Keyword: High temperature NaCl

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Cloning of $\beta$-Glucosidase Gene from Streptomyces coelicolor A3(2) and Characterization of the Recombinant $\beta$-Glucosidase Expressed in Escherichia coli (Streptomyces coelicolor A3(2)로 부터 $\beta$-Glucosidase 유전자 클로닝 및 재조합 효소의 특성)

  • Kim, Jae-Young;Kim, Bong-Kyu;Yi, Yong-Sub;Kang, Chang-Soo;Ahn, Joong-Hoon;Lim, Yoong-Ho
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.99-104
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    • 2009
  • The $\beta$-glucosidase gene from Streptomyces coelicolor A3(2) was cloned and expressed in Escherichia coli. The ORF consisted of 1377 nucleotides encoding 51 kDa in a predicted molecular weight. Effects of pH indicated that the $\beta$-glucosidase showed similar activity using $\alpha$-pNPG($\rho$-nitrophenyl-$\alpha$-D-glucopyranoside), $\beta$-pNPG($\rho$-nitrophenyl-$\beta$-D-glucopyranoside), and $\beta$-pNPF($\rho$-nitrophenyl-$\beta$-D-fucopyranoside) at range of pH 3 to 10, and high activity using $\beta$-pNPGA ($\rho$-nitrophenyl-$\beta$-D-galactopyranoside) from pH 5 to 10, especially, 3.3 times higher activity at pH 9. Effects of temperature indicated that the $\beta$-glucosidase showed low activity using $\alpha$-pNPG, $\beta$-pNPG, and $\beta$-pNPF from $20^{\circ}C$ to $70^{\circ}C$, and increased activity using $\beta$-pNPGA from $30^{\circ}C$ to $50^{\circ}C$, 1.8 times higher activity at $50^{\circ}C$ than at $30^{\circ}C$. According to activity determination of other substrates, the enzyme was active on daidzin, genistin, and glycitin, inactive on esculin and apigenin-7-glucose. The EDTA and DTT as reducing agents inhibited $\beta$-glucosidase activity, but SDS and mercaptoethanol did not inhibit. Monovalent or divalent metal ions such as $MnSO_4$, $CaCl_2$, KCl, and $MgSO_4$ did not inhibited $\beta$-glucosidase activity. $CuSO_4$ and NaCl showed low inhibition, and $ZnSO_4$ inhibited 3.3 times higher than control.

Cultural and Physiological Conditions for T-2 Toxin Production by Fusarium sp. (Fusarium 균주의 배양 조건 및 생리적 조건에 따른 T-2 toxin의 생성 조건)

  • 홍성희;양규환
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.91-96
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    • 2000
  • The cultural and physiological conditions for the T-2 toxin [4,15-diacetoxy-8-(3-mety1butyloxy)-12,13- epoxy-trichothec-9-en-3-01, $C_{24}H_{30}O_9$] production by Fusarium spp. were studied. Thin layer chromatography (TLC) assay and the microbiological assay uslng Rhodotomla rubra were used to quantitate tbe T- 2 toxin. Among the four strains of Fusarium spp., F tn'cinctum NRRL 3299 was best for T-2 toxin production. In solid culture, white com grit medium was best for T-2 toxm production. Temperature played a critical role in the production of T-2 toxin. T-2 toxin production was favored by long duration of low-temperature incubation. The growth and toxin production were relatively high on galactose, fructose, glucose, and sucrose media, when each was used as a sole carbon source, and relatively low on sorbitol, glycerol, and lactose media. For nitrogen sources, $NH_4^(+) and NO_3^{-}were used well as a sole nitrogen source, but $NO_2^-$ was not used. Initial pH and speed of shaker also affected the production of T-2 toxin. From temperature shifting experiment, it is clear that T-2 toxin metabolic pathway is regulated by temperature-dependent enzyme depression or enzyme induction system.

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Isolation of High Yielding Alkaline Protease Mutants of Vibrio metschnikovii Strain RH530 and Detergency Properties of Enzyme

  • Chung, So-Sun;Shin, Yong-Uk;Kim, Hee-Jin;Jin, Ghee-Hong;Rho, Hyune-Mo;Lee, Hyune-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.349-354
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    • 2000
  • Abstract A facultative alkalophilic gram-negative Vibrio metschnikovii strain RH530, isolated from the wastewater, produced several alkaline proteases (VAP) including six alkaline serine proteases and a metalloprotease. From this strain, high yielding YAP mutants were isolated by NTG treatment. The isolated mutant KS1 showed nine times more activity than the wild-type after optimization of the culture media. The production was regulated by catabolite repression when glucose was added to the medium. The effects of several organic nitrogen sources on the production of the YAP were investigated to avoid catabolite repression. The combination of 4% wheat gluten meal (WGM), 1.5% cotton seed flour (eSF), and 5% soybean meal (SBM) resulted in the best production when supplemented with 1% NaCl. The YAP showed a resistance to surfactants such as $sodium-{\alpha}-olefin$ sulfonate (AOS), polyoxy ethylene oxide (POE), and sodium dodecyl sulfate (SDS), yet not to linear alkylbenzene sulfonate (LAS). However, the activity of the YAP was restored completely when incubated with LAS in the presence of POE or $Na_2SO_4$. The YAP was stable in a liquid laundry detergent containing 6.6% SLES (sodium lauryl ether sulfate), 6.6% LAS, 19.8% POE, and stabilizing agents for more than two weeks at $40^{\circ}C$, but the stability was sharply decreased even after 1 day when incubated at $60^{\circ}C$. A washing performance test with the YAP exhibited it to be a good washing power by showing 51 % and 60% activity at $25^{\circ}C{\;}and{\;}40^{\circ}C$, respectively, thereby indicating that the YAP also has a good detergency at a low temperature. All the results suggest that the YAP produced from the mutant strain KSI has suitable properties for use in laundry detergents.rgents.

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Investigation on Bacillus anthracis isolated from Kyong-Ju (경주에서 분리된 탄저균에 대한 연구)

  • 이준규;이은미;차우양;김정화;김영환;이양수;김우현;정종식
    • Korean Journal of Veterinary Service
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    • v.18 no.1
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    • pp.41-56
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    • 1995
  • The present study was conducted to investigate results of B. anthracis isolated from Anthrax in the Kyong-Ju of Feb. 12. 1994. 1. In biochemical feature, B. anthracis was a gram-positive rod, non-motility, sporulation, capsulation. It was positive in gelatinase, starch hydrolysis, glucose. But negative in urease, arabinose, mannitol, xylose. 2. B. anthracis grew well on B4 Br A TSA after incubation for 24 hours. The organisim grew well on BA, Br. A, NA, TSA after incubation for 72 hours. The media grew well on Br A instead of BA. 3. On 5% blood agar by laboratory animal, ${\beta}$ -hemolysis was produced from 36 hours to 48 hours incubation. There was perfect ${\beta}$-hemolysis after incubation for 48 hours. On the other side ${\beta}$-hemolysis was begun on 5% goat blood agar after incubation for 60 hours. 4. In the test of antimicrobial susceptibility, B. anthracis was very sensitive to AM, CF, TE, ENR, GM, AN, DFX, S, P, TYLO, N, KM, C, E, Lins+Sp, NN, CC, CFP, CB were sensitive one by one. B. anthracis was no-sensitive to L, XNL, TIA, CL, SXT 5. B. anthracis had never sensitivity to direct inoculation of rat and chicken, after subcutanous inj. It was very sensitive to mouse and goat, hamster, guinea pig, rabbit had a sensibility one by one. 6. The dead laboratory animal which had been inoculated with B. anthracis preserved at $37^{\circ}C$ incubation, B. anthracis didn't cultivate on non-dissected animal after 80 hours but cultivate on dissected animal after 360 hours. 7. The rapidly death could cause high concentration, died from 420 after S. C. 8. The blood smeared samples of hamster from inoculation with B. anthracis, spore germinated In 37$^{\circ}C$ after 5 hours, in $32^{\circ}C$ after 6 hours, in room temperature after 9 hours, in $-4^{\circ}C$ to $-20^{\circ}C$ after 10 hours. 9. B, anthracis inoculated to laboratory animal after SC or PO. Mice and rats feces didn't cultivated with B. anthracis after SC, but did cultivated with B. anthracis after PO. 10. In the test of disinfectant, B. anthracis was high effective to $HgC1_2$, formalin, effect phenol, cresol, but non-effect NaOH, ethanol.

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Hydrothermal Gold mineralization of the trabong district, vietnam : Mineralogical and geochemical study (베트남 짜봉(Trabong) 지역의 열수 금 광화작용 : 광물 및 지화학적 연구)

  • 한진경
    • Economic and Environmental Geology
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    • v.32 no.2
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    • pp.129-139
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    • 1999
  • Hydrothermal gold deposits of the Trabong district in Vietnam occur as single-stage quartz $\pm$ calcite veins (0.3-1.2 m thick) which fill fault fractures in graphite-bearing gneiss and schist of the Chulai Complex and Kham Duc Formation of the Proterozoic age. Ore grades are 1.3 to 92.4 g/ton Au. Ore mineralogy is very simple, consisting mainly of pyrite with minor amounts of base-metal sulfides and electrum. Gold grains occur in two assemblages as follows: (1) early, Fe-rich (7.2-10.4 mole % FeS) sphalerite + electrum (50.4-64.3 atom % Au) assemblage occurring as inclusions in pyrite; (2) late, Fe-poor «4.7 mole % FeS) sphalerite + galena + electrum (47.6-81.7 atom % Au) assemblage occurring along fractures of pyrites. Based on fluid inclusion data and thermochemical considerations of ore mineral assemblages, ore minerals were formed at high temperatures (about $230^{\circ}C$ to $420^{\circ}C$) from $H_{2}O-CO_{2}(-CH_{4})$-NaCI fluids with the sulfur fugacity of about $10^{-6}$ to $10^{-10}$ atm. Fluid inclusion data also indicate that ore mineralization occurred mainly as a result of fluid unmixing accompanying $CO_2$ effervescence. Calculated oxygen and measured hydrogen isotope compositions of mineralizing waters (${\delta}^{18}O_{V-SMOW}$ values = 5.3 to 8.6$\textperthousand$, ${\delta}D_{V-SMOW}$ values = - 60 to - 52$\textperthousand$), along with the sulfur isotope compositions of vein sulfides (${\delta}^{34}S_{CDR}$ values = - 1.2 to 2.8$\textperthousand$) and carbon isotope compositions of inclusion $CO_2$ (${\delta}^{13}C_{PDB}$ values = - 4.7 to - 2.0$\textperthousand$) indicate that the high temperature (mesohypothermal) gold mineralization formed from a magmatic fluid.

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From food wastes into useful probiotics: Development of Lactic acid bacteria as useful probiotics for animal feed without antibiotics using food waste (음식물류폐기물의 자원화를 위한 항세균 활성 유산균의 개발)

  • Seo, Jeong-Yong;Song, In-Geun;Lee, Dae-Gyu;Lee, Ki-Young;Kim, Young-Jun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.14 no.2
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    • pp.112-120
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    • 2006
  • For the conversion of food waste into a good recycling material such as animal feed without antibiotics, thirteen lactic acid bacteria, which can be used as good probiotics for animal feed, were isolated from the intestine and feces in pigs. All isolates showed strong tolerance to high salt (4% of NaCl), acid(pH4.0), and bile juice (0.8% of oxgall). The growth rate was best at $37^{\circ}C$ in all strains. Among the isolates, Lactobacillus plantarum CJY-22, L. brevis CJY-42, L. arizonensis CJY-3, and Pediococcus sp. CJY-41 showed higher and broader spectrum of antimicrobial activities against six different pathogens such as Salmonella, typhimurium. L. plantarum CJY-22 has also grown well at $25^{\circ}C$, making this strain as an appropriate candidate for the fermentation of food waste at room temperature, thus conducting the fermentation process cost-effectively.

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Identification of genes related to ER stress in bZIP28 gene transgenic potato plant

  • Kim, Dool Yi;Kim, Kyung Hwa;Choi, Man Soo;Ok, Hyun Choog;Kim, Jae Hyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.153-153
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    • 2017
  • Potato (Solanum tubersosum L.) is susceptible to various environmental stresses such as frost, high temperature, and drought. Enhancement of potato drought tolerance can reduce yield loss under drought that has negative effect on potato tuber growth. Genetic engineering can be utilized to achieve this goal, but such approaches using endogenous potato genes have rarely been applied. Since unpredictable global weather changes cause more severe and frequent water limiting conditions, improvement of potato drought tolerance can minimize such adverse effects under drought and can impact on sustainable potato production. Genetic engineering can be utilized to improve potato drought tolerance, but such approaches using endogenous potato genes have rarely been applied. We were obtained AtbZIP28 gene transgenic potato plants. It is identified transcript levels at various stress conditions, polyethylene glycol (PEG), NaCl, abscisic ${\underline{acid}}$ (ABA). Also, For identification to regulate ER stress response genes in AtbZIP28 gene transgenic potato plant, we screened seven potato genes from RNA-seq analysis under TM treatment. Five and two genes were up- and down-regulated by TM, respectively. Their expression patterns were re-examined at stress agents known to elicit TM, DTT, DMSO and salt stress.

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Electrochemical Behavior of Plasma Electrolytic Oxidized Films Formed in Solution Containing Mn, Mg and Si Ions

  • Lim, Sang-Gyu;Choe, Han Cheol
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2017.05a
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    • pp.80-80
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    • 2017
  • Titanium and its alloys that have a good biocompatibility, corrosion resistance, and mechanical properties such as hardness and wear resistance are widely used in dental and orthopedic implant applications. However, they do not form a chemical bond with bone tissue. Plasma electrolytic oxidation (PEO) that combines the high voltage spark and electro-chemical oxidation is a novel method to form ceramic coatings on light metals such as tita-nium and its alloys. This is an excellent re-producibility and economical, because the size and shape control of the nano-structure is relatively easy. Silicon (Si), manganese (Mn), and magne-sium (Mg) have a useful to bone. Particularly, Si has been found to be essential for normal bone, cartilage growth, and development. Mn influences regulation of bone remodeling be-cause its low content in body is connected with the rise of the concentration of calcium, phosphates and phosphatase out of cells. Pre-studies have shown that Mg plays very im-portant roles in essential for normal growth and metabolism of skeletal tissue in verte-brates and can be detected as minor constitu-ents in teeth and bone. In this study, Electrochemical behavior of plasma electrolytic oxidized films formed in solution containing Mn, Mg and Si ions were researched using various experimental in-struments. A series of Si-Mn-Mg coatings are produced on Ti dental implant using PEO, with the substitution degree, respectively, at 5 and 10%. The potentiodynamic polarization and AC impedance tests for corrosion behav-iors were carried out in 0.9% NaCl solution at similar body temperature using a potentiostat with a scan rate of 1.67mV/s and potential range from -1500mV to + 2000mV. Also, AC impedance was performed at frequencies anging from 10MHz to 100kHz for corrosion resistance.

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Feasibility Tests on Struvite Production from Liquid Fertilizer by Utilizing Ferronickel Slag and Sewage Sludge Ash (페로니켈슬래그와 하수슬러지소각재를 이용한 액비로부터 스트루바이트 생산 타당성 연구)

  • Kim, Hyeon;Kwon, Gyutae;Jahng, Deokjin
    • Journal of Korean Society on Water Environment
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    • v.34 no.3
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    • pp.316-327
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    • 2018
  • Liquid fertilizers made from livestock manure contain high concentrations of nitrogen and phosphorus and thus are used as a fertilizer. However, excessive use of liquid fertilizer causes eutrophication of agricultural land and nonpoint source pollution. In this study, as a means of lowering the nutrient concentrations, struvite ($MgNH_4PO_4{\cdot}6H_2O$) production from the liquid fertilizer was investigated. When liquid fertilizers produced in Gyeonggido were analyzed, its characteristics differed by region and season, but the phosphorus concentration was commonly lower than that of nitrogen. When $K_2HPO_4$ and $MgCl_2$ were added to the liquid fertilizers, the optimal pH for struvite formation was pH 9.5. For environmentally friendly sources of magnesium and phosphate, ferronickel slag (FNS) and sewage sludge ash (SSA) were suspended in deionized water and extracted by sulfuric acid with various mass ratios. The optimum conditions for extracting FNS and SSA were 4.0 M sulfuric acid and 0.35 mass ratio of sulfuric acid to sewage sludge ash, respectively. For forming struvite, 0.233 L of SSA leachate (SSAL) was added into 0.3 L of liquid fertilizer containing 2,586 mg/L of ammonia and 110 mg/L of phosphate, pH was then adjusted to pH 9.5 using 10 M of NaOH. Afterwards 0.333 L of FNS leachate (FNSL) was added to this mixed solution. After a reaction for 1 hr at room temperature, the remaining concentrations of magnesium, ammonium, and phosphate were less than 50 mg/L, 500 mg/L and 150 mg/L, respectively, and 30 g of precipitates were obtained, most of which were struvite.

Purification and Characterization of an Intracellular NADH: Quinone Reductase from Trametes versicolor

  • Lee, Sang-Soo;Moon, Dong-Soo;Choi, Hyoung-T.;Song, Hong-Gyu
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.333-338
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    • 2007
  • Intracellular NADH:quinone reductase involved in degradation of aromatic compounds including lignin was purified and characterized from white rot fungus Trametes versicolor. The activity of quinone reductase was maximal after 3 days of incubation in fungal culture, and the enzyme was purified to homogeneity using ion-exchange, hydrophobic interaction, and gel filtration chromatographies. The purified enzyme has a molecular mass of 41kDa as determined by SDS-PAGE, and exhibits a broad temperature optimum between $20-40^{\circ}C$, with a pH optimum of 6.0. The enzyme preferred FAD as a cofactor and NADH rather than NADPH as an electron donor. Among quinone compounds tested as substrate, menadione showed the highest enzyme activity followed by 1,4-benzoquinone. The enzyme activity was inhibited by $CuSO_4,\;HgCl_2,\;MgSO_4,\;MnSO_4,\;AgNO_3$, dicumarol, KCN, $NaN_3$, and EDTA. Its $K_m\;and\;V_{max}$ with NADH as an electron donor were $23{\mu}M\;and\;101mM/mg$ per min, respectively, and showed a high substrate affinity. Purified quinone reductase could reduce 1,4-benzoquinone to hydroquinone, and induction of this enzyme was higher by 1,4-benzoquinone than those of other quinone compounds.