• Journal of Plant Biology
• /
• v.39 no.1
• /
• pp.41-48
• /
• 1996

Two forms of homo serine dehydrogenase have been isolated from 8-day-old cotyledons of Canavalin lineata by a heat denaturation, ammonium sulfate fractionation, DEAE-8ephacel ion exchange and Sephacryl 8-300 gel filtration chromatographies, and Pro cion red dye, Cibacron blue dye and Resource Q column chromatographies. The molecular weights of T -form (threonine-sensitive) and K-form(threonine- insensitive) were estimated to 230 kD and 135 kD, respectively. In the presence of 10 mM threonine, the activity of T-form was inhibited with almost 70%, but that of K-form was not at all. The Km values tor homo serine of T- and Kform were 1.6 mM and 0.3 mM, respectively. The Km values for NAD of T- and K-form were 2.34 mM and 0.03 mM, respectively. And Km values for NADP of two isozymes were the same as 0.01 mM. The activities of T- and K-form were markedly stimulated up to 4.9and 2.8-fold, respectively, by 400 mM KCI. The partial purified(gel filtration) enzymes(Tform and K-form) can be reversibly converted.verted.

• KSBB Journal
• /
• v.18 no.2
• /
• pp.122-126
• /
• 2003

Multi-cycle chromatographic separation of Iysozyme from egg white was investigated. Multi-cycle chromatography was performed by repeated cycling(one cycle: resin equilibration, sample loading, washing, elution). Two types of cation exchange resins, Cellufine CM C-200 and Bio-rex 70, were used to determine the optimum condition for the separation of Iysozyme by multi-cycle chromatography. The resin was equilibrated in 20 mM Na-phosphate buffer(pH 7.0). Chromatograms of UV absorbance levels of every cycle were compared to confirm the eluting ability of Iysozyme in the two types of gel. Collected samples from eluting regions in every cycle were assayed by 15% SDS-PAGE.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.32 no.8
• /
• pp.1323-1327
• /
• 2003

Persimmon has been considered to have therapeutic values for various diseases in Korea. Dried persimmon has been applied to wounded parts for anti-inflammatory and analgesic activities. Anti-coagulant fraction from Persimmon stem was purified through gel filtration, phenyl Sepharose, DEAE-Sephadex and additional gel filtration column chromatographies. Its molecular weight was estimated to be 130,000 ∼ 180,000. By element analysis, its main components were C, H, and O. The anti -coagulant was heat- stable and completely inhibited after periodate oxidation, indicating that it was a complex carbohydrate.

• Polymer Science and Technology
• /
• v.13 no.3
• /
• pp.384-392
• /
• 2002

• Journal of the Korean Chemical Society
• /
• v.23 no.3
• /
• pp.165-174
• /
• 1979

A purification technique of glucose oxidase was developed. Using the gluconyl-${\omega}$-aminohexyl Sepharose affinity chromatography, it was partially purified 14.6 folds with 79.7% yield. With the combination of the affinity chromatography and Sepharose 6B gel filtration, the enzyme was purified 27.2 folds from the broth with 74.1% yield. The final purified preparation showed 90.83 U of glucose oxidase activity per mg of protein and a single band by 7% polyacrylamide gel electrophoresis. The absorption spectrum and substrate specificity of the enzyme were studied and the fianal preparation showed the optimal pH between 5.6 and 6.0, the optimal temperature at $40^{\circ}C$, $8.5{\times}10^{-3}M$ of $K_m$ for D-glucose, and 3.43 kcal/mole of the activation energy.

• Journal of Ginseng Research
• /
• v.23 no.4
• /
• pp.217-221
• /
• 1999

We have isolated an antithrombin active polysaccharide in red ginseng by procedures comprising three major steps involving alkaline extraction, anion exchange and gel permeation chromatography. Active polysaccharide behaved as a single band on cellulose acetate membrane electrophoresis. The average molecular mass was estimated to be about 177 kDa by gel filtration. This polysaccharide was found to be an acidic heteropolysaccharide that contains uronic acid moiety $(40.2\%)$, sulfate group $(9.2\%)$ and protein $(1.5\%)$ in addition to neutral sugar consisted of rhamnose, mannose, galactose, arabinose, glucose, fucose and xylose in a molar ratio of 1.00 : 0.88 : 0.86 : 0.78: 0.70 : 0.33 : 0.22. This polysaccharide inhibited blood coagulation via the intrinsic pathway like heparin in a dose-dependent manner. The clotting of fibrinogen by thrombin was also mitigated by the presence of this polysaccharide.

• The Korean Journal of Food And Nutrition
• /
• v.10 no.3
• /
• pp.309-313
• /
• 1997

Bench scale Sikhyes were produced from rice and glutinous rice and limit dextrins in rice Sikhye and glutinous rice Sikhye were purified by ethanol precipitation and Biogel P-2 gel chromatography and FPLC on Superose 12 column and analyzed. The purified limit dextrin in rice Sikhye and glutinous rice Sikhye showed bot signal of $\alpha$-1,4- and $\alpha$-1,6-glucosidic linkage with its estimation ratio of 4.5:1 and 5.9:1, respectively, by 1H-NMR analysis. Limit dextrins were hydrolyzed by pullulanase. The enzyme hydrolysis products contained maltose, maltotriose, maltotetraose, maltopentaose and matohexaose. These results suggest that limit dextrins were composed of these maltoolgosaccharide series with $\alpha$-1,6-glucosidic bond.

• Korean Journal of Food Science and Technology
• /
• v.21 no.4
• /
• pp.486-491
• /
• 1989

The textural properties of cooked rice and the molecular weight distribution of amyloses of three rice cultivars different in cooking quality were investigated by the method involving rheometry and gel chromatography he tested rice cultivals were Chunmabyeo (short grain), Yongmunbyeo (medium grain) and Mahatma (long grain) The cooked rice prepared with Mahatma was higher in hardness and lower in adhesiveness than those prepared with the other two cultivars. The molecular wright distribution of amylose from Mahatma was found to be higher than those from the other cultivars.

• Korean Journal of Microbiology
• /
• v.43 no.2
• /
• pp.83-90
• /
• 2007

Sphingomonas sp. KS 301, which was isolated from oil contaminated soil, was shown to have five different SODs (SODI, II, III, IV, V) which can be separated by DEAE-Sepharose chromatography, and SOD III was finally purified in this study by ammonium sulfate precipitation, DEAE-Sepharose chromatography, Superose 12 gel filtration and Uno-Q1 ion exchange chromatography. The molecular weight of SOD III was 23 kDa as determined by SDS-PAGE and the apparent molecular weight of the native enzyme was estimated to be approximately 71 kDa by Superose-12 gel filtration chromatography. These data suggest that the purified SOD consists of at least two subunits. The specific activity of the SOD III was higher than Mn type or Fe type SOD of Escherichia coli by 5 fold. To determine the type of SOD III, inhibitory effects of $NaN_{3},\;H_{2}O_{2},\;KCN$ were examined. 10 mM $NaN_{3}$ was able to inhibit 56% of the SOD III activity, which indicates that this SOD is Mn type. The optimum pH of the SOD III was 7.0 and the optimum temperature was $20^{\circ}C$. N-terminal amino acid sequence of purified SOD III was most similar to those of Psudomonase ovalis and Vibrio cholerae among bacteria.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.24 no.5
• /
• pp.273-288
• /
• 1991

To elucidate the physiological and biochemical differences between chondrichthyes and osteichthyes, the properties of the specific digestive enzymes in cat-shark, Cephaloscyllium umbratile, and mackerel, Scomber japonicus, were studied. Homogenous trypsin proved through the disc-electrophoresis, SDS-PAG electrophoresis and gel filtration was obtained from the pancreas of cat-shark by $50-70\%$ saturated ammonium sulphate fractionation, DEAE-Sephadex A-50 column chromatography, benzamidine-Sepharose 6B affinity chromatography and Sephadex G-75-120 gel filtration. Two types of trypsins were also obtained from the pyloric caeca of mackerel by $30-70\%$ saturated ammonium sulphate fractionation and the slightly modified procedure from the method adopted in the purification of cat-shark trypsin. The two trypsins, designated trypsin A and B, were proved their homogeneity by disc- and SDS-PAG electrophoresis and gel filtration. The molecular weights of the trypsins were estimated to be 31,700 for cat-shark trypsin, 30,000 for mackerel trypsin A and 29,000 for mackerel trypsin B by SDS-PAG electrophoresis, but those were estimated to be 21,500 for cat-shark trypsin, 23,700 for mackerel trypsin A and 21,500 for mackerel trypsin B by gel filtration. The trypsins exhibited their optimum conditions at pH 9.0 and on temperature ranged from $45^{\circ}C\;to\;50^{\circ}C$ for cat-shark, and at pH 8.0 and a temperature of $50^{\circ}C$ for mackerel trypsin A and B, respectively. The cat-shark trypsin was stable at pH 10.0 and the temperature below $10^{\circ}C$, whereas the mackerel trypsin A and B, were stable in the range over pH 7.0 to pH 9.0 below $10^{\circ}C$ and at pH 8.0 below $35^{\circ}C$, respectively. The mackerel trypsins were severely inhibited by some heavy metal ions such as $Ag^{2+},\;Cu^{2+}\;and\;Hg^{2+}$ compared to cat-shark trypsin. All of the enzymes were also inhibited by antipain, leupeptin, TLCK(tosyllysine chloromethyl ketone) and SBTI(soybean trypsin inhibitor) remarkably. The inhibitory effects of PMSF(phenylmethane sulphonylfluoride), DFP(diisopropyl fluorophosphate) and benzamidine were indicated that these enzymes belong to serine-proteases.