• Title/Summary/Keyword: DEAE-cellulose chromatography

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Fractionation and Physicochemical Characteristics of Caprine Casein by DEAE-Cellulose (DEAE-Cellulose에 의한 산양유 Casein의 분별 및 이화학적 성질에 관한 연구)

  • Im, Dong-Hyeon;Jeon, U-Min;Han, Gyeong-Sik;Kim, Byeong-Cheol;Hwang, Gwang-Yeon;Kim, Se-Heon
    • Journal of Dairy Science and Biotechnology
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    • v.18 no.1
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    • pp.1-8
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    • 2000
  • This experiment was carried out to study fractionation and physicochemical characteristics of caprine casein. Acid caseins obtained from caprine colostral and normal milk were analyzed by chymosin treatment and fractionated by DEAE-cellulose column chromatography with linear gradient and electrophoresis. Protein, fat and lactose of caprine normal milk were 2.70${\pm}$0.27%, 3.82${\pm}$0.51%, and 4.10${\pm}$0.29%, respectively. More non-protein nitrogen(NPN) was released by chymosin treatment from caprine colostral casein than normal casein. The electrophoretic pattern of caprine casein was not similar to that of bovine casein, Caprine normal casein was fractionated by DEAE-cellulose column chromatography with a 0.08${\sim}$0.18 M NaCl linear gradient into 5 pes with the proportion of 5.27%, 26.07%, 25.97%, 30.40% and 12.29%, respectively. In order to identify the pure fraction, the chymosin-treated caprine normal casein was fractionated by DEAE-cellulosecolumn chromatography with a 0.08${\sim}$0.18 M NaCl linear gradient into 6 peaks with the proportion of 17.06%, 9.10%, 17.85%, 20.11%, 27.03% and 8.85%, respectively.

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Purification of $\beta$-Galactosidase from Alkalophilic Bacillus sp. YS-309 (호알카리성 Bacillus sp. YS-309로부터 $\beta$-Galactosidase의 정제)

  • 유주현;윤성식
    • Microbiology and Biotechnology Letters
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    • v.17 no.6
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    • pp.587-592
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    • 1989
  • A strain of alkalophilic Bacillus sp. YS-309 capable of producing large amount of $\beta$-galactosidase has been isolated from soil sample. Intracellular $\beta$-galactosidase was purified 6.9 folds by procedures including ammonium sulfate precipitation, DEAE-cellulose chromatography, gel-filtration, DEAE-Sephadex A-50 chromatography with over-all yield of 17.8%. The molecular weight of native enzyme was 205, 000 by HPLC, and SDS-polyacrylamide gel electrophoresis showed that the enzyme consisted of 4 identical subunits with a molecular weight of 56, 000.

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Studies on the Constituents of Bordetella bronchiseptica. Purification and Properties of the Fractions Obtained by DEAE-cellulose Column Chromatography (Bordetella bronchiseptica의 균체성분(菌體成分)에 관한 연구(硏究) 특(特)히 DEAE-cellulose Chromatography에 의한 분획정제(分劃精制)에 대하여)

  • Kang, Byong Kyu
    • Korean Journal of Veterinary Research
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    • v.13 no.1
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    • pp.47-52
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    • 1973
  • An attempt was carried out to purify the cell components of Bordetella bronchiseptica by the mild procedures. From the supernatant of sonic treated colls, the K-agglutinogen, hemagglutinogen and skin necrotizing factor were separated by a successive use of DEAE-cellulose column chromatography and ammonium sulfate precipitation. The specific activity of purified K-agglutinogen was demonstrated by a gel-diffusion test and was also found to be free from other biologically active substances of B. bronchiseptica: namely hemagglutinin, skin-necrotizing factor and O-antigen.

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Studies on Naringinase of Mold - Part 2. Purification of Aspergillus Naringinase - (사상균 Naringin 분해효소에 관한 연구 - 제 2 보 Aspergillus 속 Naringin 분해효소의 정제에 관하여 -)

  • Ki, Woo-Kyung;Kim, Jong-Kyu;Kim, Myung-Chan
    • Korean Journal of Food Science and Technology
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    • v.5 no.2
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    • pp.78-83
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    • 1973
  • The naringin hydrolyzing enzyme has been purified from the culture filtrate of the mold Aspergillus S-1 which selected to remove the bitter test of the orange or citrus fruits industrily. In a view of purity naringinase was more effectively purified in order of molecular sieving on Sephadeex G-200, starach gel electrophoresis, chromatography or a DEAE-Cellulose column and fractional precipitation by ammonium sulfate. The purified enzyme is homogeneous in paper electrophoresis from a culture filtrate by treatment fractional precipitation with ammonium sulfate, DEAE-Cellulose treatment and Sephadex-200 column chromatography and it hydrolyse only naringin to purunin.

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Purification of Odontoglossum Ringspot Virus by DEAE-Cellulose Chromatography (DEAE 셀루로오즈 컬럼 크로마토그래피 기법에 의한 Odontoglossum 윤문 바이러스의 정제)

  • 이철호;박종오;정효원;나용준
    • Korean Journal Plant Pathology
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    • v.14 no.6
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    • pp.559-562
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    • 1998
  • Odontoglossum ringspot virus (ORSV) was finally purified from ORSV-infected orchid plants by diethylaminoethyl (DEAE) cellulose anion exchange column chromatography. The virus was reliably eluted by potassium chloride at the concentration from 0.1 M to 0.13 M. Partial purification was done by solubilization with Triton X-100 (allkylphenoxypolyethoxy ethanol) and precipitation with polyethylene glycol (PEG; MW 8,000). The finally purified ORSV represented one distinct homogeneous band and the molecular weight of its capsid protein was about 17,500 Dalton in electrophoretic analysis. Electron microscopy showed not only intact particles ranged from 280 nm to 340 nm in length, but also segmented particles that final 140 nm to 220 nm and even disks. Enzyme-linked immunosorbent assay (ELISA) showed that final yield was 12 mg/100 g of the infected leaves. Bioassay demonstrated that the purified ORSV had the normal infectivity to orchid plants and Nicotiana glutionsa. Based on these data, anion exchange column chromatography could be efficiently applied to the purification of ORSV and other viruses similar to ORSV.

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Extraction and Separation of Protein-bound Polysaccharide by Lentinus edodes (표고버섯 배양액으로부터 단백다당류의 추출 및 정제 방법)

  • 박경숙;이별나
    • The Korean Journal of Food And Nutrition
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    • v.10 no.4
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    • pp.503-508
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    • 1997
  • The extraction and separation methods of protein-bound polysaccharides from the mycelium and culture broth of L. edodes were investigated. The use 2% solution of surface active agent, Triton X-100 was effective for extraction of the protein-bound polysaccharide from the mycelium. The extraction of the protein-bound polysaccharides from mycelium with hot water was achieved by 4 hours extraction at 10$0^{\circ}C$. For the separation and partial purification of the protein bound polysaccharides the column chromatography using DEAE-Cellulose, DEAE-Sephadex and Sephadex proved to be effective.

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Purification of Porcine Leukocyte Lysosomal Hydrolases (Porcine Leukocyte Lysosomal Hydrolase의 정제에 관(關)한 연구(硏究))

  • Cho, Moo-Je
    • Applied Biological Chemistry
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    • v.20 no.1
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    • pp.136-141
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    • 1977
  • DEAE-Cellulose, Sephadex column chromatography and polyacrylamide gel electrophoresis were used to purify acid phosphatase, aryl sulfatases, ${\beta}-glucuronidase$ and cathepsin D in n-butyl alcohol extracts of porcine leukocyte Iysosomes. The degree of purification was quite high for all enzymes studied and some could be identified by histochemical reactions.

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Identification of a Bacterium which Produced D-Glucose Isomerase and Partial Purification on the Enzyme (포도당 이성화효소 생산균의 동정 및 그 효소의 부분정제)

  • Rhee, In-Koo;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.8 no.2
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    • pp.125-133
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    • 1980
  • A microorganism which produced D-glucose isomerase was identified to be similar to Streptomyces antibioticus on the morphological, cultural and physiological characteristics except the spore chain and the utilization of sucrose. D-xylose grown cells of Streptomyces sp. strain K-17 were disrupted by grinding with sea sand. D-glucose isomerase was partially purified with the fractionation by ammonium sulfate, Mn-treatment, DEAE-cellulose column chromatography, DEAE-sephadex (A-50) column chromatography and gel filtration of sephadex G-200. The enzyme was purified about 380 fold with 25 % recovery.

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Extraction and Separation of Protein-bound Polysaccharide Produced by Coriolus versicolor (Fr) Quel (구름버섯 배양액으로부터 단백다당류의 추출 및 정제방법)

  • Park, Kyung-Sook;Lee, Jae-Yang;Lee, Sang-Jik;Kim, Seon-Hee;Lee, Jae-Sung
    • The Korean Journal of Mycology
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    • v.20 no.1
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    • pp.72-76
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    • 1992
  • The extraction and separation methods of protein-bound polysaccharides from the mycelium and culture broth of Coriolus versicolor (Fr) Quel were investigated. The use of 2% solution of surface active agent Triton X-100, was effective for extraction of the protein-bound polysaccharides from the mycelium. For the separation and partial purification of the protein-bound polysaccharides, the column chromatography using DEAE-Cellulose and DEAE-Sephadex proved to be effective.

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Purification of Inositol Triphosphate Kinase from Bovine Brain (소의 뇌로부터 Inositol Triphosphate Kinase의 정제)

  • Kim, Jung-Hye;Lee, Jae-Tae
    • Journal of Yeungnam Medical Science
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    • v.13 no.1
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    • pp.46-58
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    • 1996
  • Inositol 1,4,5-triphosphate($InsP_3$) is a second messenger for mobilizing intracellular $Ca^{2+}$. It can be dephosphorylated by soluble and particulate forms on $InsP_3$ 5-phosphatase, or phosphorylated to produce inositol 1,3,4,5-tetrakisphosphate($InsP_3$) by $InsP_3$ 3-kinase. These enzymes represent possible targets for the regulation of the $InsP_3/InsP_4$ signal. $InsP_3$ 3-kinase which catalyses th ATP-dependent phosphorylation of $InsP_3$ was purified from bovine brain tissue. All operation were carried out at $4^{\circ}C$. Fresh tissure was homogenized and centrifuged. The supernatant was pooled. Proteins were precipitated from 10% polyethylene glycol, and suspended solution was applied to DEAE cellulose column for chromatography. As the result of above procedure, two isozymes of $InsP_3$ 3-kinase, I and II were obtained. Each isozyme was applied to Matriz green gel, Calmodulin-Affigel 15 column and subsequent phenyl-TSK HPLC column. Specific activites(SA) and fold of puriety were observed at each purification step of chromatography. At DEAE cellulose chromatography, SA were I, 0.6 and II, 4.8 nM/min/mg, and folds were I, 17.2 and II, 16.6. At Matrix green gel chromatography, SA were I, 18 and II, 11 nM/min/mg, folds were I, 62.1 and II, 38.0. At calmodulin-Affigel 15 column chromatography, SA were I, 19 and II, 13 nM/min/mg, folds were I, 65.5 and II, 44.8. Finally $InsP_3$ kinase I and II were purified 3,103-fold and 2,310-fold, and SA were I, 900 and II, 670 nM/min/mg, respectively. SDS-polyacrylamide gel electrophoresis elucidated 3 distinct fractions of Mr of 145,000, 85,000 and 69,500 from isozyme I, and 2 distinct fractions of Mr of 79,000 and 57,000 from isozyme II.

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