• Title/Summary/Keyword: $NH_2$ column

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Purification and Properties of Alkaline Pretense from Xanthomonas sp. YL-37 (Xanthomonas sp. YL-37 균주가 생산하는 Alkali성 단백질분해효소의 정제 및 성질)

  • 장형수;권태종
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.427-434
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    • 1998
  • An alkaline protease was 4-fold purified, yielding 2.3% of recovery by ammonium sulfate precipitation, CM-cellulose column chromatography and Sephadex G-100 column chromatography. The purified enzyme was estimated to be monomeric with molecular weight of about 62,000 from polyacrylamide gel eletrophoresis (PAGE) and sodiumdodecylsulfate polyacrylamide gel electrophoresis (SDS-FAGE). The optimal pH and temperature of the alkaline pretense activity were 11.0 and 50$^{\circ}C$, respectively, exhibiting high stability at pH value from 6.0 to 11.0 at 50$^{\circ}C$ for 30 minute. The alkaline pretense was activated by MnSO$_4$, CaCl$_2$, and was inhibited by CuSO$_4$, ZnSO$_4$, HgCl$_2$, EDTA and EGTA. Also, the enzyme was found to be a metaloenzyme requiring Mn$\^$2+/ as cofactor. The NH$_2$-terminal amino acid of alkaline protease was alanine. The Km and Vmax values of this enzyme for casein was 4.0 mg/$m\ell$ and 5,500 unit/$m\ell$, respectively.

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Comparison of QuEChERS and Solid Phase Extraction for Accurate Determination of Pesticide Residues in Kimchi Cabbage and Strawberry using Isotope Dilution Mass Spectrometry

  • Seonghee Ahn;Kebede Gebeyehu Mekete;Byungjoo Kim
    • Mass Spectrometry Letters
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    • v.14 no.4
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    • pp.178-185
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    • 2023
  • QuEChERS is used worldwide as a universal sample preparation method with many benefits, such as being quick, easy, cheap, effective, rugged and safe. This study examined whether QuEChERS can be employed in isotope dilution mass spectrometry (ID-MS) for accurate analysis of pesticides in food. The ratios of fortified values and measured values of malathion and fenitrothion using the QuEChERS method were compared with those using the solid phase extract (SPE) method which was previously used in this laboratory. The separations of the two pesticides on DB-5MS and VF-1701MS columns were compared. Malathion and fenitrothion were fortified into kimchi cabbage and pretreated with the QuEChERS method and the SPE method. The results obtained using the DB-5MS column varied according to the sample preparation method, column and pesticide level. Using the VF-1701 column, ratios were 98-102% by both QuEChERS and Carb/NH2 SPE method for all fortification level. Malathion and fenitrothion were fortified into strawberry samples for comparison with kimchi cabbage. The results for the strawberry samples indicated that the ratios were not influenced by the sample preparation methods or GC column. The QuEChERS method could be acceptable in the ID-MS method for pesticide residue analysis in food, however other conditions should be carefully considered for accurate determination, such as the column, amount of analyte and food matrix.

Fate of Nitrogen and Phosphorous in Hydroponic Waste Solution Applied to the Upland Soils (시설하우스 폐양액의 토양 처리에 따른 질소 및 인의 이동)

  • Yang, Jae-E.;Park, Chang-Jin;Yoo, Kyung-Yoal;Kim, Kyung-Hee;Ok, Yong-Sik
    • Korean Journal of Environmental Agriculture
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    • v.24 no.2
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    • pp.132-138
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    • 2005
  • Objective of this research was to evaluate the fate of nitrogen and phosphorous in hydroponic waste solution from the plastic film house cultivation applied to the upland soil by column leaching and field experiment. The pH and EC of leachate were decreased by the reaction with the upland soil in the column leaching experiment. The EC and concentrations of $H^+,\;K^+,\;and\;{NH_4}^+$ of leachate were decreased as the column length (soil depth) was increased. But these were increased as the amounts of the hydroponic waste solution were increased field experiment growing red pepper (Capsicum annum L.) to monitor the nutrients movement using ion exchange resin capsule demonstrated that the nutrient concentration of soil solution was increased in the orders of $PO_4-P. Nitrate concentration of resin capsule inserted into the soil was relatively higher than other nutrients $(NH_4-N\;and\;PO_4-P)$ at the 45 cm of soil depth. The overall results demonstrated that the hydroponic waste solution could be recycled as plant nutrients to enhance fertility of soils. But nitrate leaching was a major factor for safe use of the hydroponic waste solution in soil.

Purification of Antifungal Antibiotic NH-B1 from Actinomycete NH 50 Antagonistic to Plant Pathogenic Fungi (식물병원진균에 길항효과가 있는 방선균 균주 NH50에서 항진균성 항생물질 NH-B1의 순수 분리)

  • 김현겸;김범석;문석식;황병국
    • Korean Journal Plant Pathology
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    • v.14 no.3
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    • pp.191-202
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    • 1998
  • About 300 actinomycetes were isolated from two forest and one sea-shore soil and tested for inhibitory effects on mycelial growth of six plant pathogenic fungi Magnaporthe grisea, Alternaria mali, Colletotrichum gloeosporioides, Phytophthora capsici, Fusarium oxysporum f. sp. cucumerinum, and Rhizoctonia solani. Among 300 actinomycetes tested, only 16 actinomycetes showed the antifungal activity against the test fungi. Isolate NH 50 was selected for production and purification of antifungal antibiotic substances. Actinomycete isolate NH 50 displayed the broad antifungal spectra against 11 plant pathogenic fungi. To identify actinomycete isolate NH 50, cultural characteristics on various agar media, diaminopimelic acid type, and morphological characteristics by scanning electron microscopy were examined. As a result, actinomycete isolate NH 50 was classified as a rare actinomycete that had LL-DAP type and did not produce spores. After incubation of isolate NH 50 in yeast extract-malt extract-dextrose broth, antifungal compound NH-B1 that inhibited mycelial growth of some plant pathogenic fungi was purified from the methanol eluates of XAD-16 resins by a series of purification procedures, i.e., silica gel flash chromatography, C18 flash chromatography, Sephadex LH-20 column chromatography, silica gel medium pressure liquid chromatography (MPLC), C18 MPLC, and high pressure liquid chromatography (HPLC). UV spectrum and 1HNMR spectrum of antifungal compound NH-B1 dissolved in methanol were examined. The antibiotic NH-B1 showed the major peaks at 230 and 271.2nm. Based on the data of 1H-NMR spectrum, NH-B1 was confirmed to be an extremely complex polymer of sugars called polysaccharides. The antibiotic NH-B1 showed strong antifungal activity against Alternaria solani and Cercospora kikuchi, but weak activity against M. grisea.

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Purification and Assay of Extracellular Autolysin from Moraxella sp. CK-l (Moraxella sp. CK-1의 세포외 Autolysin의 분리 정제 및 활성도 측정)

  • 오영상;이장현;한명수;윤문영
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.148-154
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    • 2003
  • Moraxella sp. CK-l is known to inhibits the growth of Anabaena cylindrica, a cyanobacterium. It has been documented that the ability of this growth inhibition of Anabaena cylindrica was attributed to extracellular autolysin from Moraxella sp. CK-l. However, it remains to be elucidated identification and characterization of autolysin have yet been elucidated. In this study, we tried to purify and identify autolysin secreted from Moraxella sp. CK-l. Cells were grown in a complex liquid medium (BGC-11) and culture supernatants were collected, followed by ammonium sulfate fractionation. Fractions were further separated with anion exchange column, Mono-Q, in FPLC system and analyzed by SDS/PAGE. The fraction containing high autolysin activity showed a single distinct protein peak in anion column and molecular mass of about 17 kDa in SDS/PAGE. Nterminal amino acid sequencing of the protein was analyzed, of which result showed the homology with some proteases, including extracellular serine protease, Dichelobacter nodosus.

Mass Transfer and Hydraulic Kinematic Character Using Lattices Packings by Countercurrent Flow of Gas-Liquid Phase in Packing Column (격자형 충진물을 이용한 충진탑내 기액상의 역류에 의한 물질전달과 수력학적 동특성)

  • Kim, Jang-Ho;Ha, Sang-An
    • Journal of Ocean Engineering and Technology
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    • v.2 no.2
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    • pp.130-137
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    • 1988
  • This thesis introduced that character of a treatment technique for a mading synthetic resin Hifiow-Ring. The material system of packings make an experiment air$NH_{3}$/air$H_{2}SO_{4}$, $SO_{2}$-air/NaOH, $NH_{3}$-air/$/H_{2}SO_{4}$ under general conditions. Lattices packing compared with conventional packings was proved low pressure loss and high separation efficiency for high loading per trans unit. And an inflow materal tested for absorption and rectification, it made an experiment under a range regular temperature, low energy and small amount of money. That made possible in simple equation, volume material tranfer coefficient$\beta_{L}$ . a by absorption or $\beta_{V}$ .a calculated in all range loading. The peculiarity pressure loss $\Delta\;P/NUT_{ov}$ for Hiflow-ring contributed to a fall cost of energy, a grade number of a vacuum rectification and absorption calculation.

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Loss of Some Cationic Zeolites and Changes of Hydraulic Conductivity in a Vertical Sand Column (수직(垂直) Sand Column 내에서의 수종염기 Zeolite의 유실(流失)과 수리전도도(水理傳導度)의 변화(變化))

  • Kang, Shin-Jyung;Choi, Jyung
    • Korean Journal of Soil Science and Fertilizer
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    • v.18 no.4
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    • pp.352-358
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    • 1985
  • Loss of cationic zeolites saturated with different ions, sodium, potasium, ammonium, magnesium and calcium ions, and changes of hydraulic conductivity were investigated in labortory. The 0.5 to 1mm sands and the zeolite particles passed 0.1mm sieve were used in the column tests. The amount of zeolite particle loss was in order of Na- > K- > $NH_4-$ saturated zeolites, and little loss was observed from Mg and Ca saturated zeolites. The loss of zeolite particles was mainly due to dispersion in solution, which was prominant when 0.01N dilute mono ionic solution and distilled water were percolated. The results also showed that the changes in hydraulic conductivity were greater in the columns where the loss of dispersed zeolite particles was higher.

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Origin and Spatial Distribution of Organic Matter at Gwangyang Bay in the Fall (추계 광양만의 유기물 기원과 분포 특성)

  • Lee, Young-Sik;Kang, Chang-Keun;Choi, Yong-Kyu;Lee, Sang-Yong
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.12 no.1
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    • pp.1-8
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    • 2007
  • Environment factors related to the distribution of organic matter in surface seawater and sediments were investigated to estimate main pollution sources and range of their influence in Gwangyang Bay. The main pollution sources for the factors that affect organic matter distribution could be divided into three main sources: fresh water runoffs from Seomjin and Dong River, Gwangyang-si domestic sewage, and Yosu Industrial Complex. Considering the characteristics in horizontal distributions of the environmental factors in water column, sediment, and water current regime, the influencing range of these main sources was likely to be divided into three areas within the bay as follows: Area I receiving lots of fresh water from Seomjin River, Area II receiving lots of domestic sewage from Gwangyang-si and fresh water of Dong River, Area III receiving lots of materials from Yosu Industrial Complex. Area I seems to be characterized as low salinity, high concentration of $NO_3-N,\;and\;SiO_2-Si$, Area II as low salinity, high concentration of $NO_3-N,\;NH_4-N,\;and\;SiO_2-Si$, and Area III as high water temperature, high concentration of $NH_4-N,\;and\;PO_4-P$ in water column, high concentration of $NH_4-N,\;PO_4-P,\;and\;SiO_2-Si$ in surface sediments.

Purification and Characteristics of Tyrosinase Inhibitor Produced by Actinomycetes F-97 (방선균 F-97이 생산하는 Tyrosinase 저해제의 정제 및 특징)

  • Bang, Byung-Ho;Rhee, Moon-Soo;Kim, Jin-O;Yi, Dong-Heui
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.153-158
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    • 2008
  • An actinomycetes F-97 producing tyrosinase inhibitor was isolated from soil samples. Isolation and purification of tyrosinase inhibitor produced by F-97 was performed as follows: IRC-120 ($NH_4^+$ type) column chromatography, silica gel column chromatography, $C_{18}$ column chromatography and Sephadex LH-20 column chromatography were used successively after the centrifuged supernatant was adjusted to pH 4.0. To identify the purity of the inhibitor, octadecylsilyl(ODS) HPLC was carried out with 5% methanol as a mobile phase. Finally, the purification yield of a tyrosinase inhibitor was 5.24%. The inhibitor was very soluble in water, methanol and ethanol but insoluble in acetone, butanol, ethylacetate and chloroform. The ${\lambda}_{max}$ value of this inhibitor in water was 194nm under UV light. The biochemical test of the inhibitor was positive in Molish, Benedict, cone. $H_2SO_4$, and $KMnO_4$ tests but negative in iodine, ninhydrin, Million, Sakaguchi, xanthoproteic and Emerson tests. The tyrosinase inhibitor was stable against heat treatment of $100^{\circ}C$ for 50 minutes and pH $4{\sim}9$. The $IC_{50}$ value of this inhibitor was $19.2{\mu}g/ml$ for mushroom tyrosinase. In $1,000{\mu}g/ml$ inhibitor concentration, inhibition zone was 27 mm for Streptomyces bikiniensis NRRL B-1049. The inhibition of F-97 against mushroom tyrosinase was competitive with tyrosine.

Isolation and Quantitative Analysis of Inosine as an Index Component from Larva of Allomirina dichotoma (장수풍뎅이 유충으로부터 지표성분(Inosine)의 분리 및 정량분석)

  • Lee, Jin-Hee;Song, Myoung-Chong;Yang, Hye-Jung;Bang, Myun-Ho;Chung, Dong-Jun;Kim, Young-Chai;Kim, Se-Young;Rho, Young-Deok;Chung, Sung-Hyun;Baek, Nam-In
    • Applied Biological Chemistry
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    • v.49 no.4
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    • pp.334-338
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    • 2006
  • A repeated silica Rel column chromatography of n-BuOH fraction from the larva of Allomirina dichotoma afforded a purified nucleic acid, which was identified as inosine on the basis of the spectroscopic data including IR, MS and NMR. The HPLC analysis of the index component using silica based $NH_2$-column exhibited the contents in the hot air-dried and Far-IR-dried larvae of Allomirina dichotoma as 0.003% and 0.055%, respectively.