• YAKHAK HOEJI
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• v.41 no.5
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• pp.638-646
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• 1997

The steroid 9${alpha}$-hydroxylase activity has been detected in cytosol fraction, $100,00{\times}g$ supernatant of cell free extract of Mycobacterium fortuitum. The activity was not linear with protein concentration in the assay suggesting 9${alpha}$-hydroxylase is a multicomponent enzyme. The 9${alpha}$-hydroxylase system was partially purified through fractional saturation of ammonium sulfate, strong anion exchange (Mono Q) column chromatography, gel filtration (Superose 12) column chromatography, and testosterone affinity gel chromatography. Ammonium sulfate 50~60% saturated fraction of the cytosol gave 9${alpha}$-hydroxylase activity. For further purification, the half-saturated ammonium sulfate fraction was applied to Mono Q, Superose 12, or affinity gel column. The purification factors of 9${alpha}$-hydroxylase containing fraction after Mono Q, Superose 12, and affinity gel chromatography was 13, 11, and 17 respectively.

• 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.

• Korean Journal Plant Pathology
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• v.10 no.3
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• pp.215-221
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• 1994

Polygalacturonase (PG) produced by Botrytis cinerea in the culture broth containing citrus pectin as a carbon source was partially purified and characterized. PG was produced on a range of carbon sources such as starch, glycerol, cellobiose, and Na+-PAG with total activities of 34.8, 32.0, 29.2, 27.8 units, respectively. The specific activity was highest with 2316.7 units on Na+-PGA. Proteins of culture filtrate were concentrated with polyethylene glycol and acetone and applied to a hydroxyapatite column. Among three active fractions collected from the column, the reaction containing the highest PG activity was resolved by a Q-sepharose column. The active fraction from the Q-sepharose column was further purified by HPLC Mono Q column. The partially purified enzyme was analyzed by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Among a few protein bands revealed, the amount of the protein of which molecular weight estimated to be 43 kDa coincided with the PG activity. The partially purified PG had optimal temperatures between 35~55$^{\circ}C$ and pH between 4.5~5.5.

• Microbiology and Biotechnology Letters
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• v.13 no.1
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• pp.65-70
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• 1985

The production media, partial purification and the enzymatic characteristics of laccase from Pleurotus ostreatus were investigated. Among various media tried the double strength onion media showed much higher production of laccase compared to glucose and/or CMC added media. The laccase from Pleurotus ostreatus has the optimum pH of 5.94 for the activity and appears to be stable at relatively broad pH spectrum (4.7-8.7). The experiments on the temperature stability shows that above 90% activity could be preserved after holding at $30^{\circ}C$ for 40 min., 60% activity was remained after 40 min. at $50^{\circ}C$. The Km value of the laccase from Pleurotus ostreatus was estimated to be 3.209mM and the molecular weight of laccase was estimated to be 55,000 by SDS-Polyacrylamide gel electrophoresis.

• Archives of Pharmacal Research
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• v.16 no.4
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• pp.295-299
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• 1993

Human renal dipeptidase (RDPase) was purified from surgically removed kdneys of renal stone aptients by affinity chromatography using its specific inhibitor, cilastain, as the ligand. The partial purified RDPase of 6 mg exhivited specific activity of 99.4 unit/mg with 2, 029 fold purification. it was composed of a slow moving major band (96%) and a fast moving minor band (4%). The minor band was not a contaminant as it showed a dipeptidase-specific activity. The kinetic parameters determined with glycyldehydrophenylalanine (Gdp) as synthetic substrate were Vmax, $322.6\;\mu{mol/min/mg}$ and km, 0.120 mM. This experiment provided biochemical evidences that sugically removed, nonfunctional kidneys in respect of glomerular filtration still retained high activity of renal dipeptidase.

• The Plant Pathology Journal
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• v.17 no.6
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• pp.386.4-386
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• 2001

• Proceedings of the Zoological Society Korea Conference
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• 1994.10a
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• pp.195.1-195
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• 1994

No Abstract, See Full Text

• Proceedings of the Zoological Society Korea Conference
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• 2000.10a
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• pp.280.1-280
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• 2000

No Abstract, See Full Text

• Journal of Microbiology and Biotechnology
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• v.9 no.4
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• pp.386-392
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• 1999

A large-scale culture of hepatitis A virus in human diploid MRC-5 cells was conducted. In a roller bottle culture, the virus was grown to a maximum titer in 3 weeks after infection. Over 95％ of the cell-associated virus was excreted after culturing the infected cells in suspension media without fetal bovine serum for 3 days. The cultured virus was inactivated with formalin, concentrated by ultrafiltration, and partially purified by ultracentrifugation in a non-ionic gradient medium of Renocal. Two separate peak fractions showing high anti-HAY ELISA titer were pooled and about 40％ of HAV antigen was recovered by this purification procedure. Of the partially purified vaccine, the protein pattern in SDS-PAGE and immunogenicity in mice were compared with a commercial HAV vaccine. In SDS-PAGE, the purified vaccine in this study and the commercial vaccine showed almost the same protein pattern. The seroconversion rate of the purified vaccine in mice was not different from that of the commercial vaccine. Therefore, we could prepare a good grade of HAV vaccine by a simple purification procedure although the purification itself was not completed.

• Journal of environmental and Sanitary engineering
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• v.14 no.4
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• pp.35-40
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• 1999

For application of a yellow pigment as food additives, stability of a water-soluble yellow pigment from Bacillus sp. PY123 was investigated. The yellow pigment from Bacillus sp. PY123 was purified with pH treatment, activated carbon and silica gel column chromatography. The partial purified yellow pigment appeared only one spot on silica gel TLC after 12 evaporation and under irradiation of UV 253nm at dark room. Rf value of the pigment was measured at 0.04 and 0.12 with development of a solvent mixture (Butanol : Acetic acid : water = 4 :1:5) and a solvent mixture (Isopropanol : Ammonia : Water = 9 :1: 2), respectively, The partial purified pigment appeared a white fluorescence under UV365nm irradiation. The partial purified yellow pigment had a main peak and a minor peak on HPLC using 20mM phosphate buffer(pH 7.0) at 1ml/min flow rate. The partial purified pigment was stable at heat treatment, acidic pH, oxide-reductants and surfactants.