• Current Research on Agriculture and Life Sciences
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• v.14
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• pp.111-122
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• 1996

S. chibaensis J-59 produced an extracellular xylanase in a CSL medium composed of 1.5% com steep liquor, 0.1% $MgSO_4{\cdot}7H_2O$, 0.012% $CoCl_2{\cdot}6H_2O$, and 0.15% glucose containing xylan. but it did not produce in the culture medium containing xylose. The production of enzyme reached to a maximum level (0.83 uints/ml) when bacteria were cultured in 2.5 l jar fermentor for 48hrs at $30^{\circ}C$ and pH 7.0. Furthermore, S. chibaensis J-59 produced an intracellular glucose isomerase in a medium containing xylan and/or xylose. Xylanase was purified 29-fold over the culture supernatants of S. chibaensis J-59 by ammonium sulfate fractionation, chromatography on DEAE-Sephadex A-50, and gel filtration on Sephadex G-200. The purified enzyme is a monomeric enzyme with a native molecular mass of 25 kDa and a subunit molecular mass of 25 kDa. The purified enzyme requires $Mg^{2+}$ for activity, $Ca^{2+}$, $Co^{2+}$ is not an inhibitor but inhibit by $Fe^{3+}$, $Hg^{2+}$, and $Cu^{2+}$, sodium dodecyl sulfate, N-bromosuccinide. Pattern of hydrolysis demonstrated that the xylanase was an endo-splitting enzyme able to break down birchwood xylan at random giving xylobiose, xylotriose and xylotetrose as the main end products.

• Proceedings of the Ginseng society Conference
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• 1990.06a
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• pp.79-85
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• 1990

The aqueous extract of fresh ginseng (Panax ginseng C.A. Meyer) contains a macromolecular fraction that showed mitogenic and comitogenic activities in human peripheral blood Iymphocytes. Purification of the crude extract by size (ultrafiltration, Sephadex G-200) and charge (DEAE-cellulose. DEAE-Sepharose) yielded a semi-purified fraction (DS-3). This fraction contains at least three subgroups of anionic macromolecules with apparent molecular weight greater than 600 kilodaltons. It is a glycoprotein with a large amount of glucuronic acid. It acts as a mitogen in both T and B cells of human peripheral blood lymphocytes. It could also potentiate the mitogenic action of Concanavalin A in Iymphocyte T cells. Such potentiation is not due to increased binding of Concanavalin A to the cell surface. Its mitogenic and co-mitogenic effects do depend on the presence of extracellular Ca2+.

• Bulletin of the Korean Chemical Society
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• v.19 no.2
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• pp.160-164
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• 1998

The NADH-cytochrome b5 reductase (NCBR), a mitochondrial external electron carrier, was purified from bovine heart and turnip and their properties were examined. The mitochondrial outer membranes separated were subjected to NCBR isolation through DEAE-Cellulose ion exchange, DEAE-Sephadex gel chromatography, and hydroxyapatite adsorption chromatography. These processes yielded the purification folds of 88 and 42 and the recovery percentages of 0.2%, 5.67% for turnip and bovine heart, respectively. The molecular weight of the NCBR from the two sources was estimated to be 35,000 using SDS polyacrylamide gel electrophoresis. The Michaelis constant Km and maximum velocity Vmax were determined by measuring the NADH-ferricyanide redox system as well as the NADPH-ferricyanide redox system. The kinetics showed that both NCBRs had higher affinities for NADH than artificial electron-acceptor substrate ferricyanide. Although NADPH had a lower affinity for the enzymes than NADH, this study showed the 2'-phosphate dinucleotide could be used as a substrate.

• Applied Biological Chemistry
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• v.12
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• pp.33-41
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• 1969

1. Crude cellulase extracted from wheat bran media of Chaetomium globosum with pH 7.0 McIlvaine buffer was fractionated by precipitation with ammonium sulfate and by treatment with the cellulose powder, DEAE-Sephadex A-25 and Amberite XE-65 (IRC-50) column chromatography. 2. Consquently two cellulases C-1 and C-2 were obtained by cellulose column chromatography. Cellulose C-1 was a powerful CMC-saccharifying and CMC-liquefying activity but cellulose C-2 was stronger CMC-liquefying activity compared to CMC-saccharifying activity and cellulase C-2 had smaller protein than that of cellulose C-1. And cellulose C-2 was fractionated by DEAE-Sephadex A-25 column chromatography into cellulase C-1-1 and cellulose C-1-2. 3. It can be obtained, therefore, that cellulose produced Chaelomium globosum consisted, at least, of three cellulases C-2, C-1-1 and C-1-2. 4. Cellulose C-1-1 was homogenous in the ultraviolet and the ultracentrifuge pattern. And cellulose C-1-1 had enzyme for CMC-saccharifying activity. 5. The optimum pH for the enzyme activity of cellulose C-1-1 was 4.0 in any methods of meas urement reducing sugar and viscosity. The optimum temperature was $40^{\circ}C$ in any methods. 6. The pH stability of cellulase C-1-1 was within pH 5.0 to pH 6.0 at $40^{\circ}C$ and fairly stable in acidic solution. 7. The heat stability was below $50^{\circ}C$ at pH 4.0 and complete heat inactivation of this cellulase occurred at $70^{\circ}C$.

• Journal of Life Science
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• v.17 no.10
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• pp.1321-1329
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• 2007

Deer antler tissue contains the most rapidly growing bone in the animal kingdom. Thus, it is likely that growing antler tissue is a rich source of local paracrine bone-stimulating factors. Growth factors, at least the insulin-like growth factor (IGF), control the bone-remodelling process. In this study, we tried to isolate and purify IGF-I from fresh antler tissue by the routine isolation and purification of protein. The purification involved ammonium sulfate precipitation, DEAE-Sepharose CL-60 ion-exchange chromatography, CM-Sepharose CL-6B ion-exchange chromatography, and Sephadex G-50 chromatography. Purified fractions from each step were analyzed by high-performance liquid chromatography (HPLC), SDS polyacrylamide gel electrophoresis (SDS-PACE), Dot-blot, and Western-blot methods. Furthermore, the quantification of partially purified IGF-I was calculated by enzyme-linked immunosorbent assays (ELISA) using antibody to human recombinant IGF-1. SDS-PAGE analysis of the final fraction yielded two molecular bands and the signal band was at 12 kDa on the Western-blot film. This purified IGF-I fraction showed a peak at retention time of eight min. The quantity of IGF-I in 20 g deer antler tissue as starting weight was calculated using a standard curve to be 2910 ng/ml, and total IGF-I amount is 0.291 g. The results show that IGF-I, which can be found in deer antler, can be partially purified and quantified by classic protein isolation methods.

• Korean Journal of Microbiology
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• v.21 no.2
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• pp.86-94
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• 1983

Polygalacturonase of Byssochlamys fulva was purified and characterized. Specific activity increased from 2.21 units/mg protein to 10.47 units/mg protein through $(NH_4)_2SO_4$ treatment, SephadexG-100 gel filtration, and DEAE-Sephadex ion exchange chromatography. Divalent cations, such as $Ca^{++}\;and\;Cu^{++}$, increased polygalacturonase activity greatly. Added as $10^{-3}M$ concentration, $Ca^{++}$ ion enhanced enzyme activity 9.8folds. Optimum temperature was $50^{\circ}C$ and optimum pH was 5.0. Activation energy of reaction was 8.69 Kcal/mole. Michaelis-Menten $constant(K_M)\;and\;V_{max}$ of reaction were $6.27{\times}10^{-3}mole/l\;and\;2.85{$\mu}moles/min$. Polygalacturonase of Byssochlamys fulva preferred polygalacturonic acid to pectin as substrate and was presumed as endo-type on the basis of the relationship between viscosity reduction and substrate degradation. Molecular weight of polygalacturonase was estimated as 55,000.

• KSBB Journal
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• v.13 no.3
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• pp.320-324
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• 1998

Agar degrading enzyme-agarase-was purified from the culture fluid of Cytophaga so/ ACLJ-18, by acetone precipitation, DEAE-Cellulose, Sephadex G-100 and CM-Sephadex C25 column chromatographies. The molecular weight of purified agarase was estimated to be 24,700 dalton by SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature for agarase activity were 7.0 and 40$^{\circ}C$, respectively. this agarase was stable in the pH range of 6.5 - 8.0 and 40$^{\circ}C$, and required 0.35M NaCl for optimum activity. And this agarase was inhibited by metal ions such as Ba2+, Cu2+, Co2+, Mn2+, Hg2+, Zn2+, and showed specificity on agar.

• Microbiology and Biotechnology Letters
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• v.24 no.3
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• pp.316-320
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• 1996

An alkaline proteinase produced by Yarrowia lipolytica TH65 was purified by 40-65% ammonium sulfate fractionation, DEAE-cellulose chromatography, and gel filtration with Sephadex G-100 and Sephadex G-75. The purified enzyme was shown as a single band on SDS-PAGE, and its molecular weight 31,500. Optimum temperature and pH were 40$\circ$C and 8.5-9.0, respectively, and the enzyme was stable below 40$\circ$C and in the pH range of 6-8. The enzyme was strongly inhibited by divalent ions, completely by PMSF, and partially by EDTA, EGTA, and phenanthroline. But the inhibitory effect in the presence of EDTA, EGTA and phenanthroline could be reversed by addition of Ca$^{2+}$. Thus, these results indicated that the purified enzyme was an alkaline serine proteinase (E.C. 3.4.21.14).

• Journal of the Korean Society of Food Science and Nutrition
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• v.20 no.1
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• pp.21-26
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• 1991

the Bacillus subtilis LY-353 which secretes the protease isolated from seafoods. The opti-mum culture condition for production of protease from B. subtilis LY-353 was as follows ; tem-perature 35$^{\circ}C$ pH 7.5 salt concentration 1.0% The purification steps involved ammonium sulfate fractionation DEAE-Sephadex A-50 column chromatography and Sephadex G-100 gel filtration A 7.33 fold purification and 6.55 yield of protease was obtained from culture broth, The optimum pH and temperature for the enzyme action were pH7.5 and 55$^{\circ}C$ respecti-bely.

• Parasites, Hosts and Diseases
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• v.38 no.2
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• pp.65-74
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• 2000

Toxoplasma-killing activities of mouse peritoneal macrophages activated by the extracts of Tetrahymena pyriformis (Korean and Chinese strains) were evaluated, and the active protein fractions from both strains were partially characterized by a method including chromatographies and SDS-PAGE. The first peak in Korean strain and the second peak in Chinese strain of T. pyriformis obtained by DEAE-Sephadex A-50 chromatography were most effective in the activation of macrophages to kill Toxoplasma gondii tachyzoites in vitro. Subsequent fractionations of obtained peak fractions were performed on a Sephadex G-200 gel. The first peaks fractionated from both strains of T. pyrtyomis had the highest toxoplasmacidal activities, and when subjected to the SDS-PAGE, one prominent band was visualized for each of the strains showing the same molecular weight of ca. 52.6 kDa. This active protein is suggested to be related to non-specific activation of mouse peritoneal macrophages.