• Title/Summary/Keyword: fractionation.

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Purification and Characterization of stu I Endomuclease from Streptomyces Tubercidicus (Streptomyces tubercidicus에 존재하는 stu I endonuclease의 정제와 특징)

  • 김기태;정미영;유욱준
    • Korean Journal of Microbiology
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    • v.25 no.3
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    • pp.180-183
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    • 1987
  • Stu I, type II restriction endonuclease, has been purified to homogeneity from Streptomyces tubercidicus (ATCC 25502), and its catalytic properties have been studied. For the purification of Stu I endonuclease free of nonspecific nucleases, DEAE-Sephadex (A-50), QAE-Sephadex (A-50) and Heparin-agarose column chromatography have been performed after ammonium sulfate fractionation of the crude extract. The enzyme was further purified by gel filtration using Sephadex G-100 column to obtain homogeneous form of protein. The single polypeptide species of Stu I endonuclease has a subunit molecular weight of 34,000 $\pm$ 1,000 daltons as judged by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Stu I endonuclease requires $Mg^{2+}$ ion for its activity and is maximally active at neutral pH (7.0-8.0) in the absence of NaCl.

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Purification of Three Extracellular Proteases from Alkalophilic Coryneform Bacteria TU-19 (호알칼리성 Coryeform bacteria TU-19가 생산하는 세종류의 균체외 단백질분해호소의 정제)

  • Choi, Myoung-Chul;Yang, Jae-Sub;Kang, Sun-Chul
    • Applied Biological Chemistry
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    • v.38 no.6
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    • pp.534-540
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    • 1995
  • Alkalophilic coryneform bacteria TU-19 isolated from soil extracellularly produced at least three proteases (Protease I, II, and III). Investigating the cultural conditions related to the enzyme production of this bacterial cell, the optimum pH and temperature were 10.0 and $30^{\circ}C$, respectively. In order to purify these enzymes from the 2 day culture broth ammonium sulfate fractionation, gel filtration and QAE-Sephadex column chromatography were performed step by step. And then these three proteases were purified to near homogeneity by judging from SDS-PAGE pattern, and had the molecular weights of 120, 80, and 45 kilodaltons, respectively. The optimum pH and temperature for the enzyme activity of Protease I and II were 10.5 and $45^{\circ}C$, respectively, and Protease II were 11.0 and $50^{\circ}C$. And the enzymes were completely inhibited by PMSF suggesting serine protease, but not affected by pCMB. 1,10-phenanthroline, IAA, and EDTA.

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Isolation and Identification of 3-Methoxy-4-hydroxybenzoic acid and 3-Methoxy-4-hydroxycinnamic acid from Hot Water Extracts of Hovenia dulcis Thunb and Confirmation of Their Antioxidative and Antimicrobial Activity (헛개나무 열수추출물로부터 항산화 및 항미생물 활성을 갖는 3-methoxy-4-hydroxybenzoic acid 와 3-methyoxy-4-hydroxycinnamic acid의 분리 및 동정)

  • Cho, Jeong-Yong;Moon, Jae-Hak;Park, Keun-Hyung
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1403-1408
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    • 2000
  • The methanol soluble fraction of the hot water extracts from Hovenia dulcis Thunb showed antioxidative and antimicrobial activity. The methanol fraction was successively purified with solvent fractionation, silica gel adsorption column chromatography, Sephadex LH-20 column chromatography, and octadecylsilane column chromatography. The purified active substances were isolated by high performance liquid chromatography. The isolated substances were identified as 3-methoxy-4-hydroxybenzoic acid (vanillic acid) and 3-methoxy- 4-hydroxycinnamic acid (ferulic acid) by LC-MS and GC-MS. Vanillic acid and ferulic acid showed antimicrobial activity against Gram-positive bacteria, Gram-negative bacteria and yeast. The DPPH-radical scavenging activity of ferulic acid appeared more active than that of vanillic acid. DPPH-radical scavenging concentration of ferulic acid and vanillic acid were $14\;{\mu}g/mL\;(SC_{50})$, $100\;{\mu}g/mL\;(SC_{10})$, respectively.

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Purification of Glucoamylase Produced by Rhizopus oryzae (Rhizopus oryzae가 생산(生産)하는 Glucoamylase의 정제(精製))

  • Hou, Won-Nyong;Chung, Man-Jae
    • Korean Journal of Food Science and Technology
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    • v.16 no.3
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    • pp.322-328
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    • 1984
  • These experiments were conducted to purify the glucoamylase produced by Rhizopus oryzae. Two forms of glucoamylase (GI and GII) from Phizopus oryzae were purified by $(NH_2)_2SO_4$ fractionation, acetone fractionation and successive column chromatography on DEAE-cellulose and CM-cellulose. The specific activities of GI and GII toward soluble starch were 157.6 U/㎎. protein (37.5 fold of crude extract), and 164.7 U/㎎. protein (39.2 fold of curde extract), respectively, and the yields of them were 4.3% and 3.8%, respectively. The two purified enzymes have shown a single band by polyacrylamide disc gel electrophoresis and SDS-polyacrylamide gel electrophoresis. The protein bands of their electrophoresis gel were revealed to have glucoamylase activity by iodine staining and were proved to be glycoprotein by periodic acid Schiff's staining.

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Development of Monitoring System for the LNG plant fractionation process based on Multi-mode Principal Component Analysis (다중모드 주성분분석에 기반한 천연가스 액화플랜트의 성분 분리공정 감시 시스템 개발)

  • Pyun, Hahyung;Lee, Chul-Jin;Lee, Won Bo
    • Journal of the Korean Institute of Gas
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    • v.23 no.4
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    • pp.19-27
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    • 2019
  • The consumption of liquefied natural gas (LNG) has increased annually due to the strengthening of international environmental regulations. In order to produce stable and efficient LNG, it is essential to divide the global (overall) operating condition and construct a quick and accurate monitoring system for each operation condition. In this study, multi-mode monitoring system is proposed to the LNG plant fractionation process. First, global normal operation data is divided to local (subdivide) normal operation data using global principal component analysis (PCA) and k-means clustering method. And then, the data to be analyzed were matched with the local normal mode. Finally, it is determined the state of process abnormality through the local PCA. The proposed method is applied to 45 fault case and it proved to be more than 5~10% efficient compared to the global PCA and univariate monitoring.

Application of Stable Isotope Analysis for Aquaculture Organisms (수산 · 양식 생물 연구를 위한 안정동위원소 분석 기법의 소개와 활용)

  • Eun-Ji, Won;Hee Young, Yun;Bohyung, Choi;Kyung-Hoon, Shin
    • Journal of Marine Life Science
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    • v.7 no.2
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    • pp.61-73
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    • 2022
  • Stable isotope analysis (SIA) is being used in various research fields including environmental science, ecology, biogeochemistry, forensics, and archeology. In this paper, for the purpose of enhancing applications and utilizations stable isotope analysis techniques to aquaculture research, we would like to introduce the background knowledge necessary to utilize stable isotope analysis techniques. In particular, with a focus on the approach using natural abundance, the principle of fractionation (change in isotope ratio) that occurs in the process of the integration of elements into biological tissues and how stable isotope ratios are determined by fractionation. This paper is intended to suggest whether SIA is used as a valuable tool in the fields of ecology and environmental science. With the understanding of the field of stable isotopes through this paper, various applications of stable isotope ratios are expected in fisheries science and aquaculture research in the future.

Geochemical and S isotopic studies of pollutant evolution in groundwater after acid in situ leaching in a uranium mine area in Xinjiang

  • Zhenzhong Liu;Kaixuan Tan;Chunguang Li;Yongmei Li;Chong Zhang;Jing Song;Longcheng Liu
    • Nuclear Engineering and Technology
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    • v.55 no.4
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    • pp.1476-1484
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    • 2023
  • Laboratory experiments and point monitoring of reservoir sediments have proven that stable sulfate reduction (SSR) can lower the concentrations of toxic metals and sulfate in acidic groundwater for a long time. Here, we hypothesize that SSR occurred during in situ leaching after uranium mining, which can impact the fate of acid groundwater in an entire region. To test this, we applied a sulfur isotope fractionation method to analyze the mechanism for natural attenuation of contaminated groundwater produced by acid in situ leaching of uranium (Xinjiang, China). The results showed that δ34S increased over time after the cessation of uranium mining, and natural attenuation caused considerable, area-scale immobilization of sulfur corresponding to retention levels of 5.3%-48.3% while simultaneously decreasing the concentration of uranium. Isotopic evidence for SSR in the area, together with evidence for changes of pollutant concentrations, suggest that area-scale SSR is most likely also important at other acid mining sites for uranium, where retention of acid groundwater may be strengthened through natural attenuation. To recapitulate, the sulfur isotope fractionation method constitutes a relatively accurate tool for quantification of spatiotemporal trends for groundwater during migration and transformation resulting from acid in situ leaching of uranium in northern China.

Study on the size-based separation of nano to micron particles in natural water and soil using flow and sedimentaion Field-flow fractionation (흐름 및 침강 장-흐름 분획법에 의한 자연수 및 토양 중 나노 크기로부터 마이크론 입자들의 크기별 분리에 관한 연구)

  • Eum, Chul Hun;Kang, Dong Young;Lee, Tae Woo;Lee, Seungho
    • Analytical Science and Technology
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    • v.22 no.1
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    • pp.75-81
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    • 2009
  • A flow and sedimentation field-flow fractionation method has been used to characterize colloidal particles in environmental samples. The opposed flow sample concentration (OFSC) method was employed. The OFSC procedure was optimized for the analysis of particles in ground water with respect to various experimental parameters including sample introduction time, flow rates, etc. The effectiveness in low concentration and characterization of the OFSC-FlFFF was demonstrated with GW-1 and GW-2 ground water samples. Ground water of upto 100 mL has been successfully loaded, concentrated, and characterized by OFSC-FlFFF. The OFSC technique allow the application of FlFFF possible for the separation and characterization of colloidal particles in very low concentrations. The results show FFF provides a simplified alternative to existing off-line concentration procedures, and shows a high potential for the applications to the analysis of dilute colloidal particles in the environmental samples.

Accelerated Fractionation In The Treatment of Brain Metastasis From Non-Small Cell Carcinoma of The Lung (비소세포성 폐암환자의 뇌전이에 대한 급속분할조사법)

  • Hong, Seong-Eon
    • Radiation Oncology Journal
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    • v.12 no.2
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    • pp.165-173
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    • 1994
  • Purpose : Metastatic cancer to the brain is a major problem for the patients with bronchogenic carcinoma, and most of these patients have a limited survival expectancy. To increase tumor control and / or to decrease late morbidity with possible shortening in over-all treatment period, multiple daily fraction technique for brain metastasis was performed. The author reperesented the results of accelerated fractionation radiotherapy in patients with brain metastases from non-small cell lung cancer. Materals and Methods : Twenty-six patients with brain metastases from non-small cell lung cancer between 1991 and 1993 received brain radiotherapy with a total dose of 48 Gy, at 2 Gy per fraction, twice a day with a interfractional period of 6 hours, and delivered 5 days a week. The whole brain was treated to 40 Gy and boost dose escalated to 8 Gy for single metastatic lesion by reduced field. Twenty-four of the 26 patients completed the radiotherapy. Radiotherapy was interupted in two patients suggesting progressive intracerebral diseases. Results : This radiotherapy regimen appears to be comparable to the conventional scheme in relief from symptoms. Three of the 24 patients experienced nausea and or vomiting during the course of treatment because of acute irradiation toxicity. The author observed no excessive toxicity with escalating dose of irradiation. An increment in median survival, although not statistically significant(p>0.05), was noted with escalating doses(48 Gy) of accelerated fractionation(7 months) compared to conventional treatment(4.5 months). Median survival also increased in patients with brain solitary metastasis(9 months) compared to multiple extrathoracic sites(4 months), and in patients with good performance status(9 months versus 3.5 months), they were statistically significant (p<0.01). Conclusion : The increment in survival in patients with good prognostic factors such as controlled primary lesion, metastasis in brain only and good perfomance status appeared encouraging. Based on these results, a multi-institutional prospective randomized trial should be initiated to compare the twice-a-day and once-a-day radiotherapy schemes on patients with brain metastasis with careful consideration for the patients' quality of life.

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Enzymatic Interesterification and Melting Characteristic for Asymmetric 1,2-Distearoyl-3-Oleoyl-rac-Glycerol Triacylglycerol Enriched Product (효소적 반응을 이용한 비대칭형 1,2-Distearoyl-3-Oleoyl-rac-Glycerol 혼합물의 생성 및 융점 특성)

  • Kim, Jin Young;Lee, Ki Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.1
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    • pp.93-101
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    • 2014
  • Asymmetric 1,2-distearoyl-3-oleoyl-rac-glycerol (SSO) triacylglycerol (TAG) is used as a cocoa butter replacer (CBR). In this study, it was produced by lipase-catalyzed interesterification of fully hydrogenated soybean oil (FHSBO) and oleic ethyl ester (OEE) in a batch type reactor at $75^{\circ}C$, 250 rpm. Different molar ratios (FHSBO : OEE=1:1, 1:2 and 1:3, w/w) and various reaction times (1, 2, 3, 4, and 5 hr) were also tested. The optimized condition for SSO was a FHSBO : OEE molar ratio of =1:1 at reaction times of 2, 3, 4, and 5 hr. Enzymatic synthesis generated SSO/SOS, as well as the other TAGs (e.g., PSO/POS, SOO/OSO, SSS), ethyl esters, monoacylglycerol (MAG), and diacylglycerol (DAG). After scale-up, fractionation by solvent (methanol and acetone) fractionation and column chromatography was applied. To reduce ethyl esters, high-melting TAGs (e.g., SSS), and SOO/OSO in reactants, solvent fractionation was applied. Using a silica gel column (sample : silica gel=2:1, wt%), MAG and DAG were removed at $25^{\circ}C$. The major fatty acid composition of the final products (with a high SSO/SOS content) was palmitic acid (C16:0, 10.9~12.9 area%), stearic acid (C18:0, 52.2~54.9 area%), and oleic acid (C18:1, 34.2~35.5 area%). In reversed-phase HPLC analysis, the major TAG species of the final product (FHSBO : OEE=1:1, 2 hr) were SSO/SOS (82.31 area%) and PSO/POS (14.51 area%). Based on the $[SS]^+$ : $[SO]^+$ ratio obtained by RP-HPLC/APCI-MS, the final product had a higher SSO (AAB type TAG) content than cocoa butter (CB). The solid fat index (SFI) of CB and the final product obtained were similar with a narrow melting point range around ~32 to $35^{\circ}C$.