• Korean Journal of Food Science and Technology
• /
• v.30 no.5
• /
• pp.1169-1178
• /
• 1998

This study was performed to purify fig pectinesterase(F-PE) and characterize its in situ activity. Three kinds of F-PE were partially separated by using ammonium sulfate fractionation, Q-Sepharose column, CM-cation exchanger column chromatography, and HPLC. One of those was anionic protein and the others were cationic proteins. All of them had approximate molecular weight of 27,000 and lost rapidly their activity during storage. Therefore alternative crude enzyme was prepared by suspending the freeze dried and milled fig powder in 0.1 M NaCl at pH 7.5. F-PE had the optimum pH of 8.5, the optimum temperature of $50^{\circ}C$ with activation energy of 7,671 cal $mol^{-1}K^{-1}$ and stability up to $55^{\circ}C$ with 10 minutes heating. Optimum activity was obtained in $0.2{\sim}0.4$ M NaCl with optimum solubility at above 0.8 M NaCl.

• Journal of Radiation Protection and Research
• /
• v.10 no.2
• /
• pp.109-112
• /
• 1985

Radioisotope tracer experiments on the distribution and the binding of radionuclides to proteins in bivalve were carried out in order to gain further information on biochemical behavior of radionuclides in marine bivalve, Gomphina melanaegis. The radioactivities (cpm/g) of $^{65}Zn\;and\;^{54}Mn$ after 7 days exposure were highly concentrated in liver and kidney in comparison to soft parts. The gel filtration profile of $^{65}Zn$ in liver and kidney showed three elution peaks, while $^{54}Mn$ showed two peaks in liver and three peaks in kidney. On the gel filtration of $^{137}Cs$ in liver and kidney, most of $^{137}Cs$ were eluted on one peak. Thus, it was considered that each radionuclide was bound to different proteins in liver and kidney of bivalve.

• Parasites, Hosts and Diseases
• /
• v.33 no.4
• /
• pp.323-330
• /
• 1995

Protease activity was identified in crude extracts of Pnrqgonimw westermnni eggs which were purified from infected dog lungs, isolated on 14 weeks after metacercarial challenge. The eggs were used after removing possibly contaminated host or worm tissues on their shell surfaces. In the crude egg extracts, high proteolytic activities against carboBfrb enzoyl - ph enylalanyl - arginyl-4- methoxy- β- naphthylamide (Cbz - phe - arg- MNA) and Azocoll were detected whereas those against succinyl-alanyl-propyl-phenylalanyl-p- nitroanilide (Suc-ala-pro-phe-pNA) were not revealed. The eVe eBdlibited the maximal activity at pH 6. Its activity was inhibited by specific cysteine protease inhibitors, 105 M I- trans-epoxysuccinylleucylamido (4-guanidino) butane (E-64) and 1 mM iodoacetamide (LAA) while potentiated by 6.5-fold in the presence of 2.5 mM dithiothreitol (DTT) . When the enzyme was purified partially by Sephacryl S-300 High Resolution gel filtration, it migrated as a single homogeneous band at 35 kDa. The 35 kDa cysteine protease has been recognized neither in the metacercariae nor in the adult. These findings indicated the presence of at least one protease of cathepsin family in immature eggs of f westernani.

• Microbiology and Biotechnology Letters
• /
• v.16 no.2
• /
• pp.92-97
• /
• 1988

Cysteinylglycinase was partially purified from Proteus mirabilis by consecutive procedure. The specific activity was increased about 16-fold to that of cell-free extract. The enzyme was found rather unstable on ammonium sulfate precipitation ann the precipitated enzyme protein became partially insoluble during dialysis. The precipitated enzyme was found to be solubilized by treatment of 4% Triton X-100 effectiviely, The optimum temperature and pH of the enzyme activity were 35$^{\circ}C$ and 7.3, respectively. After heat treatment of the enzyme at 5$0^{\circ}C$ for 30 min, it lost the activity to 70%. The enzyme was stable at pH 7.0-8.0. The molecular weight of the cysteinylglycinase was found to be about 190,000 by Sephadex G-150 gel filtration. The enzyme was activated by the addition of Mn$^{2+}$ and $Mg^{2+}$ ions. The maximal activation was obtained in preincubation with $Mg^{2+}$ ion for 30 min. The enzyme catalyzed the hydrolysis of various dipeptides and tripeptides. The Km and Vmax values for cysteinylglycine were 1.60 mM and 0.24 m unit/ mg, respectively.

• Applied Biological Chemistry
• /
• v.22 no.4
• /
• pp.191-197
• /
• 1979

${\beta}-Tyrosinase$ was purified and crystallized from cells of Escherichia intermedia A-21 grown in a medium supplemented with 0.2% L-tyrosine. Molecular weight of its subunit, Km value and absorption spectra were determined. Crystallization methods were also studied to eliminate any unnecessary procedures. The results obtained were as follows: 1. The purification procedure included ammonium sulfate fractionation, dialysis against potassium phosphate buffer, pH 6.0 and pH 7.0, and DEAE-Sephadex column chromatography. In the column chromatography, 11 mg of protein was applied per ml of DEAE-Sephadex for efficiency. 2. Steps of protamine sulfate treatment and Sephadex G-150 gel filtration could be eliminated for this enzyme from the known procedures. 3. The purified enzyme was dissolved in 0.01M potassium phosphate buffer containing 2-mercaptoethanol, with a concentration of 20mg/ml. Crystalline enzyme, which appears as hexagonal rods, was obtained by adding solid fine powdered ammonium sulfate to the enzyme solution. 4. Absorption maxima of the enzyme appeared at 340 and 430nm when associated with pyridoxal phosphate. 5. Km value of the enzyme for L-tyrosine was $2.31{\times}10^{-4}M$ and the molecular weight of its subunit was determined by SDS-polyacrylamide electrophoresis to be approximately 50,000.

• Journal of Life Science
• /
• v.18 no.6
• /
• pp.789-795
• /
• 2008

To study the possible use of probiotics in fish farming, we evaluated antagonism of antibacterial strain Bacillus amyloliquefaciens H41 against the fish pathogenic bacterium Vibrio anguillarum NCMB1. The purification of growth inhibition factor produced by B. amyloliquefaciens H41 was achieved by obtaining supernatant of this bacterium. The growth inhibition factor was purified to homogeneity by 70% ammonium sulfate precipitation, DEAE-sephadex A-50 ion exchange chromatography, sephadex G-200 gel filtration column chromatography, and sephadex G-50 gel filtration column chromatography with 40.8 fold of purification and 2.9% yield. The molecular weight of the purified growth inhibition factor was 48 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum pH and temperature for the growth inhibition factor were pH 7.5 and $30^{\circ}C$, respectively. The activity of growth inhibition factor was enhanced slightly by some metal ions, such as $Mg^{+2}$, $Mn^{+2}$, but was inhibited by the addition of $Co^{+2}$, $Hg^{+2}$, $Zn^{+2}$ and $Ag^{+2}$. NaCl stability of the growth inhibition factor was observed with 50% residual activity at 3% NaCl concentration. Toxicity test showed that the purified B. amyloliquefaciens H41 growth inhibition factor did not affect the live of Japanese flounder (Paralichthys olivaceus) and the effectiveness was 78% of residual lethality compared to commercial antibacterial agents.

• Microbiology and Biotechnology Letters
• /
• v.35 no.2
• /
• pp.163-172
• /
• 2007

This study was designed to investigate that inorganic germanium $(GeO_2)$ did not exist in germanium-fortified yeast or obtained to non-detectable value by current analytical methods and equipments. For this purpose, we achieved $GeO_2$ qualitative analysis protocol which could be the scientific basis of the study. Since reddish brown precipitate was formed from the reaction of $GeO_2$ with 1 equiv $NaBH_4$, and dark brown precipitate was also formed from the reaction of $GeO_2$ with 2 equiv $NaBH_4$, $GeO_2$ was qualitatively analyzed by observing these particular colored-precipitates. Because no color change was showed from the reaction between $NaBH_4$ and $SiO_2$, the color change could be caused by charge transfer transition on Ge-O and B binding properties. The reaction between $NaBH_4$ and germanium-fortified yeast did not show any color change and precipitate formation which meant no $GeO_2$ existed in germanium-fortified yeast. The reaction between $NaBH_4$ and supernatant specimen collected from the outside of dialysis membrane (MWCO 1,200 dalton) did not show any color change and precipitate formation. Therefore, we considered that the both germaniums in and outside of the dialysis membrane were organic germaniums. Germanium-fortified yeast which was biosynthesized organic germanium can be applied not only as a new functional material for improving health, prevention and treatment of chronic degenerative diseases including cancers, and the regulation of immune system, but also as a new materials.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.32 no.5
• /
• pp.684-688
• /
• 2003

The objective of this study was to search for the best strain as a source of L- $\alpha$-glycerophosphate oxidase (GPO) production and to establish the process technology for the purification of GPO on an industrial scale. The GPO was produced by culturing Streptococcus faecium, and purified by ammonium sulfate, DEAE-cellulose and hydroxyapatite chromatography. The relative activity was 60 units/L for 5. faecim ATCC 12755, 65 units/L for 5. faecium ATCC 19634, and 67 units/L for 5. faecium $M_{74}$.LC, respectively. The optimum condition for fermentation was $37^{\circ}C$ for temperature, 300 rpm for stir rate, 0.5 L/min for aeration rate and 17 hours. The main culture medium prepared by the modified AC medium. AC medium consists of 0.1% glucose, 0.2% glycerol, 1.0% tryptone and 1.0% yeast extract, 0.5% $K_2HP0_4$, pH 7.0. The GPO was purified by ammonium sulfate fractionation and ion exchange column chromatography, The yield and purity were 17.2% and 5.3 fold, respectively.

• Childhood Kidney Diseases
• /
• v.6 no.1
• /
• pp.102-108
• /
• 2002

We report two cases of hemolytic uremic syndrome (HUS) associated with Escherichia coli O114. Two cases were similar and showed the same clinical courses. After prodrome of diarrhea and vomiting lasting 1-2 days, azotemia persisted for about 10 days, and during that period, the patients were on peritoneal dialysis. They recovered without any sequelae after about 15 days. Direct multiplex PCR of stool culture revealed eae and stx2 gene and the result of ELISA done on the colony positive of one gene confirmed Escherichia coli O114. This is the first report of HUS associated with Escherichia coli O114. We recommend, Shiga toxin producing bacterial Infection must be considered and efforts should be made to scrutinize the organism in all diarrhea-prodrome HUS patients.(J Korean Soc Pediatr Nephrol 2002 ;6 : 102-8)

• Applied Biological Chemistry
• /
• v.46 no.4
• /
• pp.299-304
• /
• 2003

A strain YB-9 was isolated from soil as a producer of the extracellular ${\beta}-D-galactosidase$, which catalyzes the hydrolysis of lactose. The strain YB-9 was identified as Streptomyces sp. on the basis of its cultural, morphological and physiological properties. After treating culture supematant of the isolate with ammonium sulfate $(15{\sim}70%)$, the precipitated protein was used as a crude ${\beta}-galactosidase$ for analyzing its reaction properties with $para-nitrophenyl-{\beta}-D-galactoside$ $(pNP-{\beta}Gal)$ and lactose as substrates. The {\beta}-galactosidase showed its maximal activity at pH $6.0{\sim}6.5$ and $60^{\circ}C$. The hydrolyzing activity of ${\beta}-galactosidase$ for both $pNP-{\beta}Gal$ and lactose was decreased by galactose. Its hydrolyzing activity for lactose was slightly decreased by glucose, but the activity for $pNP-{\beta}Gal$ was increased to 1.3-folds by glucose. Especially, its hydrolyzing activity was not affected for lactose and was increased to 1.6-folds for $pNP-{\beta}Gal$ by xylose.