• Korean Journal of Microbiology
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• v.32 no.2
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• pp.132-138
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• 1994

L-Ascorbic acid-producing enzyme in Neurospora crassa was found to exist in mitochondria and the activity of this enzyme was increased by the addition of D-fluconno-${\gamma}$-lactone or L-gulono-${\gamma}$-lactone in the media. L-Ascorbic acid-producin enzyme in N. crassa has been purified with ammonium sulfate precipitation. DEAE Sepharose CL-6B ion exchange chromatography. Sephacryl S-200 gel filtration chromatography and Reactive yellow 3-agarose dye affinity column chromatography. The specific activity of this enzyme was increased to 239.6 fold and the yield was 2.1%. The molecular weight of the native enzyme was 150.000 dalton when it was estimated with Sephacryl S-200 gel filtration chromatography. Its molecular weight appeared as 75.000 dalton by SDS-polyacrylamide gel electrophoresis. which suggested that this enzyme was consisted with two identical subunits. The optimal pH for this enzyme was 9.0 and the $K_m$ value for D-galactono-${\gamma}$-lactone was 0.073 mM.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.313.2-313.2
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• 2002

Environmentally benign protein staining method in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using both an acidic dye, zincon (ZC) and a basic dye. ethyl violet (EV) 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 produces bluish violet colored bands. It is a rapid procedure, involving only fixin and staining steps that are completed in 45 min. (omitted)

• The Korean Journal of Mycology
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• v.25 no.2 s.81
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• pp.85-90
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• 1997

The Isozyme activities of Lentinula edodes were studied as a preliminary study for genetic analysis after protoplast fusion. The presence of peroxidase, esterase, superoxide dismutase, acid phosphatase, alkaline phosphatase, alcohol dehydrogenase and ${\alpha}-amylase$ was examined. An intracellular buffer-soluble protein from the mycelia was used for enzyme analysis on nondenaturing polyacrylamide gels. The auxotrophs of Lentinula edodes were positive for peroxidase, esterase, superoxide dismutase and acid phosphatase. However, alkaline phosphatase, alcohol dehydrogenase and ${\alpha}-amylase$ were not detected. The esterase and peroxidase were not affected by the various culture age. Isozyme identification may be a useful tool after protoplast fusion.

• BMB Reports
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• v.29 no.2
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• pp.116-121
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• 1996

In earlier studies, the genes encoding Escherichia coli thioredoxin and Corynebacterium nephridii thioredoxin C-3 were fused via a common restriction site in the nucleotide sequence coding for the active site of the proteins to generate two chimeric thioredoxins, designated E-C3 (N to C-terminal) and C3-E. The hybrid thioredoxins were overexpressed in E. coli from the cloned chimeric thioredoxin genes by a T7 promoter/polymerase system. To investigate the structure-function relationship of thioredoxin, we purified the E-C3 hybrid thioredoxin through ammonium sulfate fractionation, DEAE-cellulose chromatography, and Sephadex G-50 gel filtration. Its purity was examined on SDS-polyacrylamide gel electrophoresis and the molecular weight of the purified E-C3 hybrid thioredoxin was estimated to be 12,000. On native polyacrylamide gels, the purified E-C3 hybrid thioredoxin shows a much lower mobility than E. coli thioredoxin. E-C3 hybrid thioredoxin exhibits a 40-fold lower catalytic efficiency with E. coli thioredoxin reductase than E. coli thioredoxin. It was shown to catalyze the reduction of insulin disulfide by dithiothreitol. The purified E-C3 hybrid thioredoxin was also characterized in other aspects.

• Microbiology and Biotechnology Letters
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• v.13 no.1
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• pp.87-91
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• 1985

Hpa I endonuclease from Haemophilus parainfluenzae has been purified of homogeneity and its physical and ezymatic properties have been studied. For the purification of the enzyme, Heparin agarose, SP-sephadex C-25, DEAE-sephadex A-50 and phosphocellulose chromatography columns were used. The denatured and reduced form of the enzyme is a monomer of molecular weight of $30,000{\pm}1,000$ as judged by 10% polyacrylamide gel electrophoresis containing 0.1% sodium dodesyl sulfate. Hpa I endonuclease was maximally active at neutral pH (7.0 to 7.5) in the presence of 50 mM NaCl.

• Journal of the Korean Professional Engineers Association
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• v.18 no.2
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• pp.16-22
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• 1985

This study was carried out to evaluate the quality of soybean sprouts grown under three different sources of light. The soybean was germinated under light through blue (251 ux), red (751 ux) and white (50 to 200 lux) polyethylene films at 25$^{\circ}C$ for 10hr./day. Vitamin C, chlorophyll, cellulose and total protein contents were determined and texture was evaluated by tasting soybean sprouts soup. protein pattern in the soybean sprouts were investigated using polyacrylamide gel electrophoresis. Vitamin C, chlorophyll and cellulose contents were increased by white light intensity. The texture of the sprouts grown under white light (50 lux, 100 lux) was better than darkness, but fought under 200 lux. Vitamin C contents of soybean sprouts grown under various sources of light (in order of light : blue > white > red) were higher than theses of ones grown in the darkness. Biosynthesis of chlorophyll was not correlated to Vitamin C content. Total protein contents of cotyledon was not changed significantly under light irradiation. But the soybean sprouts grown under different quality of light, hypocotyl was higher than those grown darkness. (37% and 20% higher for blue light and white light) Densitometric tracing of disc polyacrylamide gel electrophoretic patterns showed that protein of hypocotyl under white light had more high molecular weight protein.

• Journal of the Korean Society of Food Science and Nutrition
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• v.21 no.6
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• pp.681-685
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• 1992

Low molecular weight RNA(LMW RNA : 5S rRNA and tRNAs, <150 nucleotides) profiles of several bacteriocin production lactic acid bacteria from pig meats and reference lactic acid bacteria were generated on 10% denaturing polyacrylamide gel electrophoresis. Data evaluation including three molecular weight markers enabled the calculation of relative nucleotide units(RNU) for every band. Gels profiles and RNU evaluations were effective for identification of lactic acid bacteria species. LMW RNA profiles of lactic acid bacteria showed no variation in dependence on APT(All Purpose Tryptone Broth), TSB(Tryptic Soy Broth), MRS(Lactobacilli MRS Broth) different cultural medium.

• Korean Journal of Food Science and Technology
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• v.32 no.5
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• pp.1158-1167
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• 2000

To produce ${\beta}-cyclodextrin({\beta}-CD)$, a cyclodextrin glucanotransferase(CGTase) producing Aspergillus sp. CC-2-1 was isolated from soil. The enzyme was purified and its enzymological characteristics were investigated. It was found that production of CGTase reached to the maximum when the wheat bran medium containing 0.1% albumin, 2% $(NH_4)_2S_2O_8$, 2% soluble starch and 0.2% $KH_2PO_4$ was cultured for 5 days at $37^{\circ}C$. The purity of CGTase was increased by 13.14 folds after DEAE-cellulose ion exchange chromatography and Sephadex G-100, G-150 gel filtration and the specific activity was 172.14 unit/mg. Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. The molecular weight of CGTase was estimated to be 27,800 by Sephadex G-100 gel filtration and SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature for the CGTase activity were 9.0 and $80^{\circ}C$, respectively. The enzyme was stable in pH $8.0{\sim}11.0$ at $60{\sim}80^{\circ}C$. The activity of purified enzyme was activated by $K^+,\;Cu^{2+}$ and $Zn^{2+}$. The activity of the CGTase was inhibited by the treatment with 2,4-dinitrophenol and iodine. The result suggests that the purified enzyme has phenolic hydroxyl group of tyrosine, histidine imidazole group and terminal amino group at active site. The reaction of this enzyme followed typical Michaelis-Menten kinetics with the $K_m$ value of 18.182 g/L with the $V_{max}$ of 188.68 ${\mu}mole/min$. The activation energy for the CGTase was calculated by Arrhenius equation was 1.548 kcal/mol.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.43 no.4
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• pp.259-263
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• 1998

High molecular weight glutenin (HMW-Glu) subunit compositions of 73 Korean wheat cultivars and experimental lines were evaluated by using one dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This method is suitable for obtaining a good resolution of 1Dx2 and 1Ax2$^*$ without adverse effects on separation of other HMW-Glu subunits. Korean wheats examined in this study could be divided into 15 different groups on the basis of HMW-Glu subunit compositions. From the wheat lines tested, it was identified that there were three alleles at the Glu-Al, five at the Glu-Bl and three at the Glu-D1 loci. The null allele of the Glu-Al was occurred in high frequency (79.4%), while low frequencies for 1Ax1 (12.3%) and 1Ax2$^*$(8.2%) were found. High frequency (75.3%) of the subunit pairs of 1Bx7+1By8 at the Glu-Bl loci compared with other subunits was found. The frequencies of subunits 1Dx2. 2+1Dy12 and 1Dx2+1Dy12 from the Glu-D1 loci were 54. 8% and 37.0%, respectively. However, a few Korean wheat lines (8.2%) carried 1Dx5 + 1Dy10 subunit pair which are responsible for good breadmaking quality. The information of HMW-Glu subunit compositions provide a useful tool to characterize wheat lines, and can be directly used in selection of breeding lines of different end-use properties.

• Korean Journal of Food Science and Technology
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• v.20 no.1
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• pp.90-94
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• 1988

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunodiffusion method were used for the species differentiation of yeasts, Saccharomyces cerevisiae, Candida utils, Candida tropicalis, and Kleuyveromyces fragilis. Comparing the electrophoretic patterns of soluble and membrane proteins, Saccharomyces cereνisiae was similar to Candida utilis but was different from Candida tropicalis and Kleuyveromyces fragilis. In immunochemical properties of soluble proteins, Saccharomyces cerevisiae was almost identical with Candido utilis. However, Saccharomyces cerevisiae or Candida utilis was quite different from Candida tropicalis and Kleuyveromyces fragilis in their immunoreactivities. In immunochemical properties of membrane proteins, almost the same results were obtained irrespective of four yeast species. By using SDS-PAGE and immunodiffusion methods, Saccharomyces cerevisiae and Candida utilis were difficult to differentiate but both species were easily differentiated from Candida tropicalis and Kleuyveromyces fragilis.