• 미생물학회지
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• 제20권4호
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• pp.183-188
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• 1982

Clostidium botulinum type B가 생성하는 독소를 정제할 수 있는 방법을 연구하였다. 정제과정은 독소를 ammonium sulfate로 배양액에서 침전시켜 추출한후 Polymin P를 처리하여 핵산 및 기타 단백질을 최대한 제거한 후 Sephaex G-I00에서 gel fiItration을 시키고 DEAE-Sephadex로 이온교환 크로마토그래피를 시켰다. 이러한 과정으로 정제된 독소의 회수율은 17%였으며 SDS-polyacrylamide gel electrophoresis 결과 하나의 선을 나타내 동질성을 증명하였다. 정제된 독소의 분자량은 163,000이였으며 $\beta$-mercaptoethanol을 사용하여 환원시킨 결과 분자량 106,000과 56,000의 하위 단위체로 분리되었다.

• Archives of Pharmacal Research
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• 제25권5호
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• pp.704-708
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• 2002

A fast and sensitive protein staining method in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using both an acidic dye, Coomassie Brilliant Blue R-250 (CBBR) and a basic dye, Neutral Red (NR) is described. It is based on a counter ion-dye staining technique that employs oppositely charged two dyes to form an ion-pair complex. The selective binding of the free dye molecules to proteins in an acidic solution enhances the staining effect of CBBR on protein bands, and also reduces gel background. It is a rapid staining procedure, involving fixing and staining steps with short destaining that are completed in about 1 h. As the result, it showed two to fourfold increase in sensitivity comparing with CBBR staining. The stained protein bands can be visualized at the same time of staining.

• 미생물학회지
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• 제25권1호
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• pp.1-8
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• 1987

An extracellular $\beta$-1, 3-glucanase from Pseudomonas stutzeri KF 13 was purified about 390 with 26% recovery. The purified enzyme revealed a single band by polyacrylamide gel electrophoresis and SDS-polyacrylamide gel electrophoresis. The enzyme was stable in a pH 6.0 to 9.0, and relatively thermostable. The optimal pH and temperature on the enzyme activity were found to be 5.8 and 45.deg.C, respectively. The activation energy was calculated to be 16,130 cal per mole. The Km value for laminarin was found to be 3ng per ml and the molecular weight was determined to be 28,000 by gel filtration and 26,000 daltons by SDS-acrylamide gel electrophoresis. The enzyme was inhibited by 1.0mM of $Hg^{2+}$, and strongly inhibited by 1.0mM of p-chloromercuribenzoic acid.

• 한국환경과학회지
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• 제7권6호
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• pp.853-857
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• 1998

Purification of the isocitrate lyase extracted from Microbacterium laevaniformans was investigated. The isocitrate lyase was purified 43.6 folds by the following continuous treatment with ammonium sulfate fraction, DEAE-cellulose, DEAE-sephacel and Sephadex G-200 chromatography. The purified isocitrate lyase was showed to be a single protein band by polyacrylamide gel electrophoresis. The molecular weight of the purified isocitrate lyase was estimated 54,000 Da by the SDS-polyacrylamide gel electrophoresis. The Km and Vmax values for isocitrate were estimated to be 0.83mM and 0.33units/ml, respectively. Activity of isocitrate lyase was inhibited by cystein-HCl and glutathione.

• Applied Biological Chemistry
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• 제20권1호
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• pp.136-141
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• 1977

돼지 백혈구(白血球) Iysosome의 n-butanol 추출물(抽出物)로 부터 DEAE-cellulose, Sephadex c-200 column chromatography 및 Polyacrylamide gel electrophoresis 로서 acid phosphatase, aryl sulfatase, ${\beta}-glucuronidase$로서 및 cathepsin D를 분리(分離)정제하고 그 성질을 조사하였다.

• 약학회지
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• 제28권3호
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• pp.139-147
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• 1984

L-PHA, a lectin having lymphoagglutinating activity but devoid of erythroagglutinability which is contained in Korean white kidney bean (Phaseolus vulgaris L.), was isolated and purified through several techniques such as ion exchange chromatography, hydroxyapatite column and affinity chromatography. Purified L-PHA was identified as a single band by polyacrylamide disc gel electrophoresis. The molecular weight of the L-PHA was estimated about 125, 000 daltons by polyacrylamide gel electrophoresis and it was turned out having one subunit (probably dimer of the Yachnin's model) of Mr~60, 000. Immunochemical studies also tried and these results reconfirmed the purity of this L-PHA.

• Applied Biological Chemistry
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• 제21권3호
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• pp.137-143
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• 1978

소의 갑상선에 있는 xanthine oxidase 효소도 flavin adenine dinucleotide(FAD), 모리브덴 및 철을 cofactor로서 가지고 있었으며 그 비율은 1 : 0.36 : 1.6이었다 갑상선 xanthine oxidase의 분자량은 gel filtration과 gel electrophoresis방법으로 측정하였을 때 우유에서 추출한 xanthine oxidase의 분자량과 큰 차이가 없었다. 그 효소의 활성도는 pH7.8에서 가장 높았고 isoelectric point는 electrofocusing으로 측정하였을 때 pH 6.2 였다. SDS-polyacrylamide gel electrophoresis 실험 결과는 소의 갑상선에서 추출된 xanthine oxidase가 세개의 subunit로 분해되었음을 지적했고, 최소 단위분자량 65,000정도의 polypeptide 4개로서 완전한 xanthine oxidase 한 분자를 구성하고 있을 가능성을 보였다. 갑상선 호소의 absorption spectrum을 우유에서 추출된 효소와 비교하였을 때 상당한 상이점을 나타내었다.

• 한국식품과학회지
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• 제14권1호
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• pp.1-5
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• 1982

고려인삼(Panax ginseng C. A. Meyer)중의 인베르타아제(invertase)를 연구하기 위하여 조(粗)인삼 인베르타아제를 분리 조제한 후 DEAE-cellulose 관크로마트 그래피, Sephadex G-75를 통한 젤 여과 등의 방법에 의하여 정제하였다. 정제된 인베르타아제는 specific activity가 조(粗)인베르타아제에 비해 64.7배 증가되었으며 효소활성의 recovery는 약 19.5%였다. 정제된 인베르타아제는 polyacrylamide gel electrophoresis에 의해 균일성(homogeneous)을 나타내었고 SDS-polyacrylamide gel electrophoresis에서는 2개의 subunits로 분리되었다. 두 subunits의 분자량은 각각 28,000과 20,000이었고 따라서 인삼 인베르타아제의 분자량은 48,000으로 계산되었다. 정제된 인베르타아제는 전형적인 단백질의 UV 흡수 스펙트럼을 나타내었다.

• 한국미생물·생명공학회지
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• 제16권6호
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• pp.438-442
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• 1988

The enzyme produced by Rhizopus japonicus was able to convert selectively ginsenoside-Rb$_1$which is the most abundant ginseng saponin, into ginsenoside-Rd which was known to be superior to ginsenoside-Rb$_1$pharmaceutically. The convertible enzyme was purified homogeneous from wheat bran culture of Rhizopus japonicus by ammonium sulfate fractionation and column chromatography of TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150, Sepharose 2B. Specific activity of the purified enzyme was increased to a bent 96 folds and yield was appeared to be 11% of culture extract. Evidence for homogenity was obtained from polyacrylamide and SDS-polyacrylamide gel electrophoresis. Molecular weight of the enzyme was estimated about 88, 000 daltons by Sephadex G-l50 gel filtration and SDS-polyacrylamide gel electrophoresis, and it did not consist of any subunit.

• Applied Biological Chemistry
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• 제27권2호
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• pp.73-78
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• 1984

Pseudomonas stutzeri IAM 12097의 trypticase 배지(培地)에서 36시간(時間), initial pH는 6.3일때 Exo-maltotetraohydrolase가 최대로 생산되었다. Exo-maltotetraohydrolas, 황산(黃酸)암모니아분획(分劃)과 2회(回)의 DEAE-cellulose column chromatography에 의하여 정제(精製)하였으며 정제효소(精製酵素)의 specific activity는 108.6 u/mg protein, 수율(收率)은 9.4%이었다. 본정제효소(本精製酵素)는 polyacrylamide gel electrophoresis와 SDS-polyacrylamide gel electrophoresis에 의하여 각각(各各) 단일(單一) band를 나타내었다.