• Journal of Microbiology and Biotechnology
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• v.12 no.2
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• pp.196-203
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• 2002

A novel process for urokinase purification was studied using p-aminobenzamidine as the ligand and sepharose 4B as the matrix. The adsorption, washing, and elution conditions were optimized by an unusual method. An adsorption buffer containing 2.5 M NaCl and $1\%$ Tween 80 facilitated the adsorption of urokinase on the affinity media and prevented contaminants from binding to the p-aminobenzamidine affinity gel. It was found that $5\%$ Tween 80 removed most of the contaminants from the affinity column. A 0.2 M glycine elution buffer containing 0.5 M NaCl (pH 3.0) was found to have a strong elution ability with a high recovery and purity of urokinase. A crude urokinase material of231 IU/mg protein from human urine was purified to 124,300 IU/mg protein with a purification factor of 538 and yield of $86.7\%$. As a result, a high purity urokinase was obtained with only a single affinity chromatography step. The purification process was successfully scaled-up to a 2-1 chromatography column. The resulting urokinase eluate could be directly lyophilized, thereby complying with Chinese pharmacopoeia (1995 version) standards.

• Food Science of Animal Resources
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• v.22 no.4
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• pp.343-347
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• 2002

Bovine colostrum contains various bio-functional Proteins. Especially, transforming: growth factor-${\beta}$1 (TGF-${\beta}$1) has a function in concerns with immune response. The purpose of this study was to establish the purification Processing of transforming growth factor-${\beta}$1(TGF-${\beta}$1). The highest concentration of TGF-${\beta}$1 was measured within 48 h after parturition in bovine colostrum using ELISA kit. Purification of TGF-${\beta}$1 from whey protein was carried out by the gel filtration, AF-heparin chromatography and AF-heparin rechromatography. After final purification step, TGF-${\beta}$1 with a molecular weight of 25 kD was obtained, and confirmed by silver staining and western blotting. Finally, TGF-${\beta}$1 was identified native form of 25 kD and reducing form of 12.5 kD by reducing agent.

• KSBB Journal
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• v.15 no.5
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• pp.537-540
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• 2000

In developing a HPLC purification process, it was hoped that a single chromatographic system would be sufficient to abtain pure paclitaxel in high yield. However, no such system was found, due in part to the complex taxoid profile of crude paclitaxel and to the rigorous nature of the product specification. A two step HPLC purification was adopted using reverse-phase separation on C(sub)18 as a first step, and normal-phase separation on silica as the final polishing step. Impurity profiles were established and maintained for paclitaxel, which identified and quantified each impurity observed in purified paclitaxel from these two steps, all impurities at or above 0.1% were identified. Results provide information for improving the quality control of paclitaxel production.

• Journal of environmental and Sanitary engineering
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• v.14 no.2
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• pp.32-39
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• 1999

The following is experimental result of selecting soil bacteria showing toxicity against Root-knot nematode (Meloidogyne hapla). Out of 286 strains isolated from soil, one(NC67) showing toxicity against M.hapla is selected The selected strain(NC67) is identified of B. thuringiensis subsp. indiana. It proved out that the toxic maerial against M. hapla produce by NC67 strain is an exotoxin. The result of examining the existence of the extercellular toxicity product by the toxic strain(NC67) by usign activated carbon column chromatography, Dowex 50W column chromatography and TLC of silical gel etc. proved out that it is a single material.

• Bulletin of the Korean Chemical Society
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• v.9 no.1
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• pp.15-17
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• 1988

Titanium tetrachloride is purified using adsorption column packed with activated silica gel. When 120 ml of titanium tetrachloride was passed through an adsorption column filled with 7 g silica gel, iron content in titanium tetrachloride has been reduced from 7 ppm to less than 1 ppm, and aluminum from 46 ppm to 11 ppm, while silicon content being unchanged at about 60 ppm.

• Journal of Ginseng Research
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• v.44 no.6
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• pp.784-789
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• 2020

Background: The separation of isomeric compounds from a mixture is a recurring problem in chemistry and phytochemistry research. The purification of pharmacologically active ginsenoside Rb₃ from ginseng extracts is limited by the co-existence of its isomer Rb₂. The aim of the present study was to develop an enzymatic elimination-combined purification method to obtain pure Rb₃ from a mixture of isomers. Methods: To isolate Rb₃ from the isomeric mixture, a simple enzymatic selective elimination method was used. A ginsenoside-transforming glycoside hydrolase (Bgp2) was employed to selectively hydrolyze Rb₂ into ginsenoside Rd. Ginsenoside Rb₃ was then efficiently separated from the mixture using a traditional chromatographic method. Results: Chromatographic purification of Rb₃ was achieved using this novel enzymatic elimination-combined method, with 58.6-times higher yield and 13.1% less time than those of the traditional chromatographic method, with a lower minimum column length for purification. The novelty of this study was the use of a recombinant glycosidase for the selective elimination of the isomer. The isolated ginsenoside Rb₃ can be used in further pharmaceutical studies. Conclusions: Herein, we demonstrated a novel enzymatic elimination-combined purification method for the chromatographic purification of ginsenoside Rb₃. This method can also be applied to purify other isomeric glycoconjugates in mixtures.

• YAKHAK HOEJI
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• v.38 no.5
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• pp.608-613
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• 1994

We isolated two Cd-binding high molecular weight proteins from intraperitoneally cadmium injected rat liver. Molecular weight of Cd-BP(I) purified from Sephacryl S-100 and DEAE Sepharose column chromatography and Cd-BP(II) purified from DEAE Sepharose column chromatography and Sulphonyl Sepharose column chromatography was 33,000 and 18,400, respectively. Alcohol dehydrogenase and alkaline phosphatase acitivities were not detected from two purified proteins.

• 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.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1998.11a
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• pp.161-161
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• 1998

To achieve the aim of this investigation, the extracellular protease was isolated from bacteria BAC-4, a strain was cultivated in the medium for the production of penicillin acilase in a period of 32 hours. The enzyme was first purified by aceton precipitation method, followed by ion exchange chromatography on DEAE-sephacel column. The highest specific activity of the aceton fraction was found to be 2.19 unit per mg, with degree of purification of 13 times. Further purification of the enzyme on DEAE -sephacel had a specific activity of 58.6 unit per mg and degree of purification of 344 times compared to its crude extract. The optimum pH of the enzyme was 8.4, and the potimum temparature was 37$^{\circ}C$. The K$\_$M/ and $V_{max}$ calculated at experiment conditions were found to be 0.66%(W/V) and 3.61 unit per mL respectively.

• Applied Biological Chemistry
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• v.18 no.3
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• pp.167-176
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• 1975

As a study on the ATP-inhibited ribonuclease of Bacillus subtilis the screening work for obtaining the ATP-inhibited ribonuclease negative mutant were carried out. And mutant strain was selected by the treatment of N-methyl-N'-nitro-N-nitrosoguanidine (NTG). For the selected strain the enzyme purification and some physiological properties were examined and the results obtained were as follows. 1. Among tested 1817 strains with the treatment of NTG, 101 strain was selected as a mutant strain. 2. ATP-inhibited ribonuclease was tentatively purified by several independent column chromatography. The results with Sephadex G-75 column were 30 times purification, 99% recovery, and 20 times purification, 98% recovery, respectively. 3. ATP-inhibited ribonuclease was purified by 60 times through acid treatment, ammonium fractionation, and two successive chromatography. 4. The purified ribonuclease were shown to be effectively concentrated in robonnclease content and to have reduced numbers of protein band on Disc electrophoresis. 5. This enzyme degraded single-stranded RNA to 2',3'-cyclic AMP, 2',3'-cyclic CMP, 2',3,-cyclic GMP, 2',3'-cyclic UMP and some unknown intermediates. The enzyme could not split double-stranded RNA.