• Title/Summary/Keyword: Single column

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Protease Activity from Fruit Body of Sarcodon aspratus (능이자실체의 Protease 활성)

  • Cho, Nam-Seok;Cho, Hee-Yeon
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.4
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    • pp.58-65
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    • 2004
  • This study was performed to investigate the protease activity from fruit body of Sarcodon aspratus and its features. The specific protease activity was increased with the increasing purification steps, 2.62 times by desalting, 17 times by CMC column chromatography, 113.8 times by DEAE-Sephadex A-50 column chromatography, and 728.3 times by Sephadex G-75 column chromatography. Proteases were identified as two different enzymes having different isoelectric points at pH 4.35 (its recovery rate 8%) and pH 4.7 (its recovery rate 3.5%). Those proteases were purified by 3,025 folds and 3,257 folds in terms of specific activity. Two proteases having different isoelectric points had similar enzymatic properties. This protease was estimated to be 43,000 daltons of molecular weights by SDS-PAGE. This protease with optimum pH 4 was almost stable in the pH range of 4~7. Optimal temperature of protease activity was 40 to 50℃, and the protease activity was completely inhibited at 70℃ for 30 min.

Evaluation of Removal Characteristics of Taste and Odor causing Compounds and Organic matters using Ozone/Granular Activated Carbon($O_{3}$/GAC) Process (오존($O_{3}$).입상활성탄(GAC) 공정을 이용한 맛.냄새 유발물질과 유기물질의 제거특성 평가)

  • Ham, Young-Wan;Ju, Young-Gil;Oh, Hyo-Keun;Lee, Byung-Wook;Kim, Hyun-Ki;Kim, Deok-Goo;Hong, Seung-Kwan
    • Journal of Korean Society of Water and Wastewater
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    • v.26 no.2
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    • pp.237-247
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    • 2012
  • This study assessed the removal characteristics of taste and odor causing compounds (2-methylisoborneol and geosmin) and organic matters, using a pilot-scale ozone/granular activated carbon ($O_{3}$/GAC) process treating surface water of Pal-dang reservoir in the Han river over a 3-month period. Experiments were conducted to verify the removal efficiency of $O_{3}$/GAC process which has two different empty bed contact time (EBCT) ($O_{3}$/GAC column 1 : 10 min and 2 : 15.1 min) with 10.86 min contact time of ozonation at 1.0 mg/L $O_{3}$. Spiking test using geosmin and 2-MIB was also conducted systematically to mimic the conditions when the algae appears, specifically at the levels similar to the concentrations experienced (geosmin: 250 ng/L) in the winter of 2011. In single ozonation process, organic materials, disinfection by-products (DBPs) and their precursors were disassembled but not removed completely. Meanwhile, it was verified that organic matters, taste and odor causing compounds, and DBPs were well removed when sequentially passing through the GAC process. The pilot results also showed that GAC column with larger EBCT achieved higher removal efficiency. Specifically, in spiking tests, single $O_{3}$ process showed approximately 89% removal efficiency of geosmin and 2-MIB. $O_{3}$/GAC combined process demonstrated excellent removal of geosmin and 2-MIB, which are higher than 95%.

Killer 효모 융합주 FWKS 260 이 분비하는 Killer Toxin 의 정제

  • 정기택;방광웅;우철주;정용진;김재근;송형익
    • Korean Journal of Microbiology
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    • v.30 no.3
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    • pp.160-163
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    • 1992
  • Killer toxin from killer yeast fusant FWKS 260 developed by protoplast fusion between the wild killer yeast and alcohol-fermenting yeast was purified by ammonium sulfate fractionation. Amicon PM I0 concentration. Sephadex G-200 and Scphadcx G-75 column chromatography. The purified killer toxin showed a single band by SIX-polyacvlamide gel electrophoresis. The protein part of killer toxin was active site. which was found by treating the proteolytic enzyme such as pronase E and pepsin to killer toxin. The killer toxin was stable at pH 2.0-5.0 and 20$^{\circ}$C. but inactivated with increasing temperature. The molecular weight was determined to be approximately 13.000 according to the results obtained from the SDS-polyacrylamide gel electrophoresis. It was confirmed that the purified killer toxin is glycoprotein by showing a red single band after st'tining with Schiffs reagent.

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Preparation of High-Purity Urokinase Using Single-Step Hydrophobic Interaction Chromatography with p-Aminobenzamidine Ligand

  • Cao, Xue-Jun;Zhou, Jian-Hua;Huang, Zhen-Hui;Wu, Xing-Yan;Hur, Byung-Ki
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.196-203
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    • 2002
  • A novel process for urokinase purification was studied using p-aminobenzamidine as the ligand and sepharose 4B as the matrix. The adsorption, washing, and elution conditions were optimized by an unusual method. An adsorption buffer containing 2.5 M NaCl and $1\%$ Tween 80 facilitated the adsorption of urokinase on the affinity media and prevented contaminants from binding to the p-aminobenzamidine affinity gel. It was found that $5\%$ Tween 80 removed most of the contaminants from the affinity column. A 0.2 M glycine elution buffer containing 0.5 M NaCl (pH 3.0) was found to have a strong elution ability with a high recovery and purity of urokinase. A crude urokinase material of231 IU/mg protein from human urine was purified to 124,300 IU/mg protein with a purification factor of 538 and yield of $86.7\%$. As a result, a high purity urokinase was obtained with only a single affinity chromatography step. The purification process was successfully scaled-up to a 2-1 chromatography column. The resulting urokinase eluate could be directly lyophilized, thereby complying with Chinese pharmacopoeia (1995 version) standards.

Production and Characteristics of Pullulanase from Bacillus cereus (Bacillus cereus에 의한 Pullulanase의 생산 및 특성)

  • 정만재;임계숙;조대선;우정숙
    • Microbiology and Biotechnology Letters
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    • v.20 no.4
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    • pp.409-416
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    • 1992
  • The optimum cultural temperature and time for the pullulanase production by Bacillus cereus were $15^{\circ}C$ and 72 hrs, respectively. The addition of casein, nutrient broth and egg albumin to the basal medium, respectively, increased greatly the enzyme production. The enzyme was purified by ammonium sulfate fractionation, CM-cellulose and DEAE-cellulose column chromatographies. The specific activity of the purified enzyme was 29.09 U/mg protein and the yield of enzyme activity was 17.1% The purified enzyme showed a single band on polyacrylamide disc gel electrophoresis and its molecular weight was estimated to be 61,000 by SDSpolyacrylamide disc gel electrophoresis. The isoelectric point for the purified enzyme was pH 7.0. The optimum temperature and pH were $40^{\circ}C$ and 6.5. The purified enzyme was stable below $35^{\circ}C$ and in the pH range of 6.5-11.0. It was greatly inhibited by $Ag^{+}$, $Hg^{2+}$ and $Zn^{2+}$, and its thermal stability was increased by the addition of $Ca^{2+}$ Among various substrates, pullulan was favorably hydrolyzed by the purified enzyme and the hydrolysis product 011 pulluIan was maltotriose.

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Rapid Simultaneous Determination of Anions in Mineral Water by Single Column Ion Chromatography (단일 컬럼이온크로마토그래피에 의한 광천수중 음이온의 신속한 동시정량)

  • Kim, Jong Hun;Choi, Yong Wook;Chung, Taek Kyun
    • Journal of the Korean Chemical Society
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    • v.39 no.12
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    • pp.910-917
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    • 1995
  • Analytical conditions for single column ion chromatographic determination of five anions such as F-, Cl-, NO2-, NO3-, and SO42- were optimized with respect to analysis time and separation efficiency. At the optimum condition of 2.0 mM phthalic acid at pH 4.5, five anions were effectively separated in 10 minutes so that analysis time could be reduced by 40% compared to other recommended conditions. Under this condition all the calibration curves of five anions were linear with a correlation coefficient > 0.999. Analytical results for nine commercial bottled waters were presented.

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Single step purification of potent antigenic protein from sparganum by gelatin-affinity chromatography (젤라틴 친화성 크로마토그래피를 이용한 스파르가눔 성분단백질의 순수분리)

  • Yoon Kong;Shin-Yong Kang;Seung-Yull Cho
    • Parasites, Hosts and Diseases
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    • v.29 no.1
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    • pp.1-8
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    • 1991
  • Out of many component proteins in crude saline extract of Spirometra mansoni plerocercoid (sparganum) , 36 kDa and 29 kDa proteins were found to be the most antigenic and were already purified by immunoaffinity chromatography using monoclonal antibody as a ligand. In this study, a single step purification of these potent antigenic proteins of sparganum extract was investigated. When the crude saline extract was charged to gelatin-Sepharose 4B affinity column, 36 kDa and 29 kDa protein fractions were bound. SDS-polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE/immunoblot confirmed that the bound protein to gelatin was serologically pure. When evaluated by ELISA with patients sera, the purified protein of 36 and 29 kDa also showed improved antigenicity.

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Preliminary Investigation for Feasibility of Wave Energy Converters and the Surrounding Sea as Test-site for Marine Equipment

  • Park, Jin-Yeong;Baek, Hyuk;Shim, Hyungwon;Choi, Jong-Su
    • Journal of Ocean Engineering and Technology
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    • v.34 no.5
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    • pp.351-360
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    • 2020
  • Of late, demand for test sites for marine equipment such as ASV, AUV, ROV, and various underwater sensors is increasing. The authors have focused on an oscillating water column (OWC), which is being constructed near Chagwido Island Jeju, as one of the test-sites. The main objective of the OWC is to produce wave energy and develop technologies. It has been built in the sea approximately 1 km off the coast. It has berth accommodation and some rooms that can be used as laboratories. To investigate the feasibility of its usage as a test site for marine equipment, we acquired bathymetric data around the OWC by using a multi-beam echo sounder and a single-beam scanning sonar. The accessibility of the OWC from nearby ports and the use of support vessels or ships were also investigated. 3D point cloud data from the multi-beam echo sounder and 2D acoustic images from the scanning sonar are expected to be used as references for identifying changes over time. In addition, through these experiments, we derived a procedure to use this facility as a test site by using the IDEF0 functional modelling method. Based on this preliminary investigation and previously reported examples, we determined the general conditions and preferences for evaluating the performance of various marine equipment heuristically. Finally, we developed five applications that were derived from this investigation.

Effect of unequal spans on the collapse behavior of multi-story frames with reduced beam section connections

  • Zheng Tan;Wei-hui Zhong;Bao Meng;Li-min Tian;Yao Gao;Yu-hui Zheng;Hong-Chen Wang
    • Steel and Composite Structures
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    • v.50 no.1
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    • pp.107-122
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    • 2024
  • Following an internal column failure, adjacent double-span beams above the failed column will play a critical role in the load transfer and internal force redistribution within the remaining structure, and the span-to-depth ratios of double-span beams significantly influence the structural resistance capacity against progressive collapse. Most existing studies have focused on the collapse-resistant performances of single-story symmetric structures, whereas limited published works are available on the collapse resistances of multi-story steel frames with unequal spans. To this end, in this study, numerical models based on shell elements were employed to investigate the structural behavior of multi-story steel frames with unequal spans. The simulation models were validated using the previous experimental results obtained for single- and two-story steel frames, and the load-displacement responses and internal force development of unequal-span three-story steel frames under three cases were comprehensively analyzed. In addition, the specific contributions of the different mechanism resistances of unequal-span, double-span beams of each story were separated quantitatively using the energy equilibrium theory, with an aim to gain a deeper level of understanding of the load-resistance mechanisms in the unequal-span steel frames. The results showed that the axial and flexural mechanism resistances were determined by the span ratio and linear stiffness ratio of double-span beams, respectively.

$S^{35}$(1,2-dichloro vinyl) L-Cysteine의 소에 의(依)한 대사물(代謝物)의 분리(分離)

  • Kim, Jae-Uk
    • Applied Biological Chemistry
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    • v.2
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    • pp.36-39
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    • 1961
  • It has been ascertained that S-(1, 2-dichlorovinyl) L-cysteine (DCVC) is probably the toxic component in the trichloro-ethylene extracted soybean oil meal on the bovine. For the study of the metabolites of DCVC, the components with radioactive $S^{35}$-in the urine of the calf, to which $S^{35}$-DCVC was administrated, were separated using of cellulose powder with propanol-water (70:30 V/V) which is easily removable by evaporation. As the results followings were obtained: Peak 1, which shows fractions containing single $S^{35}$-radioactive components, whose Rf is 0.815 Peak 2, which shows fractions containing jingle $S^{35}$-radioactive component, whose Rf is 0.447 Peak 3, which shows fractions containing both $S^{35}$-radioactive components whose Rfs are 0.090 and 0.171 Peak 4, which shows fractions containing single $S^{35}$-radioactive component, whose Rf is 0.142. Peak 5, which shows fractions containing single $S^{35}$-radioactive component, whose Rf is 0.084. Besides these, two of small peak were obtained. Although, the resolving power of the cellulose powder column is not sufficient, it is applicable for the study of the components of metabolytes of DCVC conveniently with the rest of removable solvent easily. Also the components with radioactive $S^{35}$ in the feces of the calf were separated using citrate buffer gradient system of Moore and stein. As the results; three $S^{35}$-radioactive components were separated.

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