• Korean Journal of Microbiology
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• v.20 no.4
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• pp.183-188
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• 1982

The neurotoxin of Clostridium botulinum type B was purified from a liquid culture. The purification steps consist of ammonium sulfate precipitation of whole culture, treatment of Polymin P(0.15%, v/v), gel filtration on Sephadex G-100 at pH5.6 and DEAE-Sephadex charomatography at pH8.0. The procedure recovered 17% of the toxin assayed in the starting culture. The toxin was homogeneous by sodium dodecyl sulfate(SDS)-polyacrylamide gel electrophoresis and had a molecular weight of 163, 000. Subunits of 106, 000 and 56, 000 molecular weight were found when purified toxin was treated with a disulfide-reducing agent and electro phoresed on SDS-polyacrylamide gels.

• Archives of Pharmacal Research
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• v.25 no.5
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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.

• Korean Journal of Microbiology
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• v.25 no.1
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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.

• Journal of Environmental Science International
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• v.7 no.6
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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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• 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.

• YAKHAK HOEJI
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• v.28 no.3
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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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• v.21 no.3
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• pp.137-143
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• 1978

Xanthine oxidase from bovine thyroid glands was found to contain FAD, molybdenum and iron in a ratio 1:0. 36:1. 6. The molecular weight of the thyroid enzyme was similar to that of the milk enzyme when estimated by gel filtration and polyacrylamide gel electrophoresis. The optimum pH for the enzyme activity was 7.8. The pH of the isoelectric point was determined to be 6.2 by electrofocusing. Sodium dodecyl sulfatepolyacrylamide gel electrophoresis experiment indicated that the enzyme was dissociated into subunits and that the molecular weight for the smallest subunit was 65,000 daltons. Absorption spectra were dissimilar between milk and thyroid xanthine oxidase.

• Korean Journal of Food Science and Technology
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• v.14 no.1
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• pp.1-5
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• 1982

An invertase from Korean ginseng (Panax ginseng C. A. Mayer) was purified by means of DEAE-cellulose column chromatography and gel-filtration through Sephadex G-75. The homogeneity of the purified invertase was proved by polyacrylamide gel disc electrophoresis. The enzyme was separated into two subunits by SDS-polyacrylamide gel electrophoresis, showing its molecular weights as 48,000. The enzyme preparation showed a characteristic protein UV-spectra.

• Microbiology and Biotechnology Letters
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• v.16 no.6
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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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• v.27 no.2
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• pp.73-78
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• 1984

The optimum culture time and initial pH, for the production of exo-maltotetraohydrolase from Pseudomonas stutzeri IAM 12097, in the trypticase medium were 36 hrs and pH 6.3, respectively. Exo-maltotetraohydrolase was purified by $(NH_4)_{2}SO_4$ and two times of column chromatography on DEAE-cellulose. Specific activity of the purified enzyme was 108.6U/mg protein and yield of the enzyme activity was 9.4%. The purified enzyme showed a single band on polyacrylamide gel electrophoresis and SDS-polyacrylamide gel electrophoresis.