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A Study on the Purification and Characteristics of Branched-Chain Amino Acid Aminotransferase in Cultural Mycelia of Cordyceps militaris (번데기동충하초 균사 중의 Branched-Chain Amino Acid Aminotransferase의 분리정제 및 그 특성에 관한 연구)

  • Kim, Sung-Tae;Park, Chung-Oh
    • Korean Journal of Clinical Laboratory Science
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    • v.37 no.2
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    • pp.78-83
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    • 2005
  • The optimum conditions of Cordyceps militaris mycelial growth, purification and characteristics of branched-chain amino acid aminotransferase [BCAT(EC 2.6.1.42)] in this mycelium were studied. Optimum pH, temperature and medium of culture of mycelia were 5.5, $22.5^{\circ}C$ and Hamada medium (HM), respectively. BCAT in homogenate of this mycelia was precipitated by 20-40% saturated solution of ammonium sulfate and then purified by DEAE (diethylaminoethyl)-Sephadex A-50 column chromatography with linear concentration gradient and Sephadex G-200 gel filtration. A single band of purified enzyme was detected on SDS-PAGE (sodium dodecylsulfate-polyacrylamide gel electrophoresis). Optimum pH and temperature of BCAT were found to be 7.8 and $29^{\circ}C$, respectively. It showed activity toward L-leucine, L-isoleucine and L-valine as a substrate. The Km values of this enzyme for L-leucine were determined to be 5.88 mM for L-leucine.

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The Production and Geochemistry of Evaporite from the Acid Mine Drainage (산성 광산배수로부터 형성되는 증발잔류광물의 생성량과 지구화학)

  • Park Cheon-Young;Cho Kap-Jin;Kim Seoung-Ku
    • Journal of the Korean earth science society
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    • v.26 no.6
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    • pp.524-540
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    • 2005
  • This study has focused on the amount of evaporites and geochemical characteritics of evaporites from the acid mine drainage and on the variation of constituents in acid mine drainage during evaporation. The various colors of evaporites are frequently observed at the rock surfaces contacting acid mine drainage. In order to produce evaporites in the laboratory, acid mine drainages were sampled from the abandoned mine areas (GTa, GTb, GH and GB) and air-dried at room temperature. During the evaporation of acid mine drainages, TDS, EC values and the concentrations of major and minor ions increased, whereas ER and DO values decreased with time. The concentration of Fe increased gradually with evaporation time in the GTb and GB, whereas GH founded in one day but rapidly not detected in the other day after due to removal of Fe by formation-precipitation of amorphous Fe hydroxide. The amounts of the evaporites were produced in amounts of 4 g (GTa), 5 g (GB), 15 g (GH), and 24 g (GTb) from 4 liter of acid mine drainage after 80 days of the evaporation, respectively. In linear analysis from the products with the parameters which are the EC, TDS, salinity, ER, DO and pH contents in field, the determination coefficients were 0.98, 0.99, 0.98, 0.88, 0.89, and 0.25 respectively. If we measure the parameters in field, it would be easy to estimate the amount of evaporites in acid mine drainage. Gypsum and epsomite were identified in all of the evaporites by x-ray powder diffraction studies. Evaporite (GTb) was heated at 52, 65, 70, 95, 150, 250, and 350oC for one hour in electrical furnaces. Gypsum, $CaSO_4\cdot1/2H_2O$ and kieserite were identified in the heated evaporite by XRD. With increased heating temperature, the intensity of the peak at $7.66/AA$ (diagnostic peak of gypsum), the peak at 5.59A ($CaSO_4{\cdot}1/2H_2O)$ and the peak at $4.83{\AA}$ (kieserite) decreased in x-ray diffraction due to dehydration. In the SEM and EDS analysis for the evaporite, gypsum of well-crystallized, radiating cluster of fibrous, acicular, and columnar shapes were observed in all samples. Ca was not detected in the EDS analysis of the flower structures of GTb. Because of that, the evaporite with flower structures is thought to be eposmite.

Physical and Chemical Properties of Coal Fly Ash Ball Substrates, the Salt Accumulation and the Effects of Washing Out Salt with Water (석탄회성형배지(Ash Ball)의 이화학적 특성과 염류집적 및 제거효과)

  • Li, Xian-Ri
    • Journal of Bio-Environment Control
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    • v.10 no.2
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    • pp.88-94
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    • 2001
  • Physical and chemical properties, the salt accumulation and leaching of salt by water of coal fly ash ball (ash ball) were evaluated in comparison with perlite and granule rockwool (rockwool). Bulk density, particle density, solid phase, and porosity of ash ball were 0.93 g.cm$^{-3}$ , 2.29 g.cm$^{-3}$ , 40.6%, 59.4%, respectively. The bulk density of ash ball was greater, while porosity was smaller, than that of perlite and rockwool. Saturation moisture capacity was 52% in ash ball, 71% in perlite, and 90% in rockwool. Water contents after drainage for 1 hr of ash ball, perlite, and rockwool were 21%, 27%, and 80%, respectively. Water content of small granules (3-5 mm) of ash ball was 5% greater than that of large (7-15 mm) grannules. The ash ball was a weak alkali substrate with pH 7.6, but not electric conductivity (EC), of the nutrient solution supplied to ash ball slightly increased. When the absorption of mineral ions to substrates were analyzed, ash ball and RW absorbed mainly PO ̄$_4$. On tomato culture, salt accumulation in ash ball substrate was similar to that in perlite. Most of the salts in the ash balls were removed by submerging the substrate eight times in distilled water. It is concluded that water holding capacity of ash ball substrate was lo as compared to other substrates, but air permeability, and water diffusion was excellent, making control of medium water content easy.

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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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Growth and Flowering Responses of Carnation (Dianthus caryophyllus) as influenced by Hydroponic Systems and Nutrient Solutions (재배방식과 양액의 종류에 따른 카네이션(Dianthus caryophyllus)의 생장과 개화반응)

  • 정순주;이범선;강종구;서범석
    • Journal of Bio-Environment Control
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    • v.6 no.3
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    • pp.190-197
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    • 1997
  • This study was conducted to determine the optimum hydroponic system and nutrient solution for promoting the growth and flowering of hydroponically grown carnation. DFT (deep flow technique) and NFT (nutrient film technique) systems were employed and four kinds of nutrient solution, which are Cooper solution, Yamasaki solution, balanced nutrient solution by Japanese Horticultural Experiment Station and PTG solution in the Netherlands were used and compared. pH was lowest in the PTG solution compared to the other nutrient solutions used. Plant height was highest in the DFT system in the plot of PTG solution as of 53.0 cm at 88 days after transplanting, while lowest in NFT system with Cooper solution. Stem diameter was more thickened in NFT system with PTG solution as of 7.2 mm at 88 days after transplanting. Days to flowering was shortened to 122.3 days in NFT system when PTG solution was used. This results suggested a combination of NFT system with PTG solution as optimum hydroponic system and nutrient solution for cut carnation production. Further trials as to the management of nutrient solution by glowing stages should be followed.

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Purification and Characterization of Lactate Dehydrogenase Isozymes in Channa argus (가물치(Channa argus) 젖산탈수소효소 동위효소들의 정제 및 특성)

  • Park, Eun-Mi;Yum, Jung-Joo
    • Journal of Life Science
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    • v.20 no.2
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    • pp.260-268
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    • 2010
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Channa argus were purified and characterized by biochemical, immunochemical and kinetic methods. The activity of LDH in skeletal muscle was the highest at 380.4 units and those in heart, eye and brain tissues were 13.4, 3,5 and 5.4 units, respectively. Citrate synthase (EC 4.1.3.7, CS) activity in heart tissue was the highest at 20.7 units. LDH/CS in skeletal muscle, heart, eye and brain tissues were 172.9, 0.6, 0.32 and 0.47. Protein concentration in skeletal muscle tissue was 14.7 mg/g and specific activities of LDH in skeletal muscle, heart, eye and brain tissues were 25.88, 0.79, 0.31 and 1.38 units/mg, respectively. Therefore, skeletal muscle tissue was anaerobic and heart tissue was aerobic. The LDH isozymes in tissues were identified by polyacrylamide gel electrophoresis, immunoprecipitation and Western blot with antiserum against $A_4$, $B_4$, and eye-specific $C_4$. LDH $A_4$, $A_3B$, $A_2B_2$. $AB_3$ and $B_4$ isozymes were detected in every tissue, $C_4$, $AC_3$, $A_2C_2$ and $A_3C$ were detected in eye tissue, and $A_3C$ was found in brain tissue. LDH $A_4$, $A_3B$, $A_2B_2$, $AB_3$, $B_4$, eye-specific $C_4$ isozymes were purified by affinity chromatography and Preparative PAGE Cells. The LDH $A_4$ isozyme was purified in the fraction from elution with $NAD^+$ containing buffer of affinity chromatography. Eye-specific $C_4$ isozyme was eluted right after $A_4$, after which $B_4$ isozyme was eluted with plain buffer. As a result, one part of molecular structures in $A_4$, $B_4$ and eye-specific $C_4$ were similar, but were different from each other in $B_4$ and $C_4$. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The activity of LDH $A_4$, $A_2B_2$, $B_4$, and eye-specific $C_4$ isozymes remained at 39.98, 21.28, 19.67 and 16.87% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The $Km^{PYR}$ values were 0.17, 0.27 and 0.133 mM in $A_4$, $B_4$ and eye-specific $C_4$ isozymes, respectively. The optimum pH of LDH $A_4$, $B_4$, eye-specific $C_4$, $A_2B_2$, $A_3B$, and $AB_3$ were pH 6.5, pH 8.5, pH 5.5, pH 6.0-6.5, pH 5.0 and pH 7.5. The $A_4$ and heterotetramer isozymes stabilized a broad range of pH. Especially, LDH activities in skeletal muscle tissue were high, resulting in a high degree of muscle activity.LDH metabolism in eye tissue seems to be converted faster from pyruvate to lactate by eye-specific $C_4$ isozyme as eye-specific $C_4$ have the highest affinity for pyruvate, and right after the conversion, oxidation of lactate was induced by $A_4$ isozyme. It was found that expression of Ldh-C, affinity to substrate and reaction time of $C_4$ isozyme were different according to the ecological environmental and feeding capturing patterns.

Changes of Tomato Growth and Soil Chemical Properties as Affected by Soil pH and Nitrogen Fertilizers (토양 pH와 질소 관비 비종에 따른 토마토 생육 및 토양화학성 변화)

  • Kang, Yun-Im;Roh, Mi-Young;Kwon, Joon-Kook;Park, Kyoung-Sub;Cho, Myeong-Whan;Lee, Si-Young;Lee, In-Bok;Kang, Nam-Jun
    • Korean Journal of Environmental Agriculture
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    • v.29 no.4
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    • pp.328-335
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    • 2010
  • This study was conducted to determine effects of soil pH and form of nitrogen fertilizers on tomato growth and chemical properties of greenhouse soil using ferigation system. Tomato (Lycopersicon esculentum Mill. cv. Superdoterang) were grown for three months in 18 L pots filled with two soil (pH 6.8 and pH 8.7). 4 different nitrogen fertilizers (urea, ammonium nitrate, ammonium sulfate, and potassium nitrate) were fertigated with different concentrations of 0, 10, 50, and 100 mg N/L during tomato cultivation. Soil pH 8.7 decreased yield and chlorophyll fluorescence compared with soil pH 6.8. Yield at soil pH 8.7 increased by ammonium nitrate and ammonium sulfate fertigation. Soil pH 6.8 induced increment of yield by nitrogen concentration than form of nitrogen fertilizers. Soil pH after cultivation of tomato decreased by application of ammonium nitrate and ammonium sulfate. Soil EC by 100 mg N/L application of ammonium sulfate was twice as much as other fertilizers. Form of nitrogen fertilizer had less effect on concentration of soil $NH_4^+$-N and $NO_3^-$-N in soil but the concentrations slightly reduced at pH 8.7. These results indicate that application of urea and ammonium nitrate for a nitrogen source of fertigation has little affects on soil chemical properties before and after tomato cultivation.

Effect of Spent Mushroom Compost on Tomato Growth after Cultivation of Button Mushroom, Agaricus bisporus. (양송이버섯 재배 후 폐상퇴비가 토마토 생육에 미치는 영향)

  • Lee, Chan-Jung;Cheong, Jong-Chun;Jhune, Chang-Sung;Kim, Seung-Hwan;Yu, Hyung-Sik
    • Korean Journal of Organic Agriculture
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    • v.17 no.1
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    • pp.83-94
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    • 2009
  • This study was carried out to investigate the effect of application of spent mushroom compost(SMC) on soil chemical properties and tomato growth. After the mushroom has been harvested, the SMC contains a lot of organic material, different microorganism and high density of mushroom hypha. SMC of white button mushroom(Agaricus bisporus) contained diverse microorganisms including fluorescent Pseudomonas sp. and actinomycetes. These isolates showed strong antagonistic to bacterial wi1t(Ralstonia solanacearum) and fusarium wi1t(Fusarium oxysporum) of tomato. The growth and sugar content of tomato showed no significant difference with other treatments by stage of maturity. The EC, exchangeable K and Ca contents of the soil during growing stage were increased in comparison to those of farmhouse practice, but available phosphate decreased. Microbial population in the soil in all growing stages showed no significant difference with other treatments, but yield of tomato decreased in some way in comparison to farmhouse practice. As the result of analysis on chemical property of soil and plant growth and yield of tomato, it seems likely that SMC of white button mushroom(Agaricus bisporus) may be used as substitute of practice compost on cultivation of tomato.

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Losses of Chemical Components by Infiltration Water during the Rice Cultivation at Silt Loam Paddy soil (영농기간 지하침투수에 의한 미사질양토 논의 화학성분 손실량)

  • Han, Kang-Wan;Cho, Jae-Young;Son, Jae-Gwon
    • Korean Journal of Environmental Agriculture
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    • v.17 no.3
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    • pp.268-273
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    • 1998
  • Changes of chemical component and losses of chemical fertilizer by infiltration water from 0.5ha of paddy field in Chinan area of Chonbuk province during the rice cultivation were investigated. The infiltration water samples were collected in a ceramic porous cup which was a buried at the 30, 50, 70, 90㎝ of soil depth. pH of infiltration water ranged $6.64{\sim}7.90$ and EC showed $324{\sim}647{\mu}$S/cm. The content of total-N, $NH_4-N$ and $NO_3-N$ were $0.58{\sim}14.59$, $0.05{\sim}4.25$, and $0.15{\sim}7.71mg/L$ respectively. The content of total-P and ortho-P were $0.009{\sim}0.077mg/L$ and $0{\sim}0.029mg/L$ The content of $Ca^{++}$, $Mg^{++}$, $Na^{+}$ and $K^{+}$ showed $0.88{\sim}4.78$, $0.22{\sim}1.04$, $0.17{\sim}0.98$, and $0.84{\sim}3.19㎎/L$. These all at the first transplanting are higher than that of other periods. The content of $SO_4^{2-}$ showed $3.92{\sim}18.72mg/L$ and decreased with a soil depth. However $Cl^-$ of infiltration water ranged $9.03{\sim}19.97mg/L$ and no difference with a soil depth. When infiltrated $2,416.5m^{3}$ of an infiltration water from 0.5ha of paddy field during the rice cultivation, losses of chemical components were 20.34㎏/㏊ of total-N, 3.54㎏/㏊ of $NH_4-N$, 10.44㎏/㏊ of $NO_3-N$, 0.16㎏/㏊ of total-P and 0.028㎏/㏊ of ortho-P. Also $Ca^{++}$, $Mg^{++}$, $Na^+$, $K^+$, $SO_4^{2-}$ and $Cl^-$ were lost 10.24, 2.84, 2.84, 7.22, 50.04 and 62.20㎏/㏊ respectively. There were lost by infiltration water 9.35% of nitrogen, 0.59% of phosphorous and 22.79% of potassium in applied chemical fertilizer.

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Relations between Soil Physicochemical Properties and Ginger Growth (토양의 물리.화학적 성질과 생강 생육과의 관계)

  • Kim, Dong-Jin;Ahn, Byung-Koo;Lee, Jin-Ho
    • Korean Journal of Organic Agriculture
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    • v.21 no.2
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    • pp.283-294
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    • 2013
  • Root-rot disease is a serious problem in ginger cultivation fields and it reduces the quality and productivity of ginger. This study was conducted to investigate the effects of different soil physical and chemical properties on the changes of ginger growth. As comparing the selected soil chemical properties after harvesting the ginger plants with those before planting them, the contents of total nitrogen and exchangeable $Mg^{2+}$ increased, whereas electrical conductivity (EC) and exchangeable $K^+$ content decreased. Potassium (K) concentrations in ginger plant were markedly higher in both its shoot and root parts ranging from 63.9 to $72.3g\;kg^{-1}$ and from 27.6 to $37.3g\;kg^{-1}$, respectively, which might be related to the decrease of exchangeable $K^+$ content in soils. Incidence rate of ginger root-rot disease in the plots ranges between 26.7% and 88.1%. It was higher in low elevation plots with clay loam soils than in high elevation plots. In addition, the incidence of the disease increased as affected by high temperature and humid condition during the growth and maturity stages of ginger. Therefore, soil texture, field slop, and drainage system as well as chemical properties should be considered to cultivate ginger plant.