• Journal of Microbiology
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• v.37 no.1
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• pp.21-26
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• 1999

Two novel calcium-binding proteins, named CAB-I and CAB-II, have been isolated from Streptomyces coelicolor. Purification of the calcium-binding proteins involved heat treatment, fractionation with ammonium sulfate, acid treatment, anion exchange and hydrophobic interaction column chromatography, FPLC gel filtration, and preparative isoelectric focusing. A chelex competitive assay and 45Ca autoradiography verified the calcium-binding ability of the proteins. The major band CAB-II has an apparent molecular weight of 26,000 determined by SDS-polyacrylamide gel electrophoresis and 340,000 determined by gel filtration. The isoelectric point of this molecule showed the acidic nature of the molecule. N-terminal amino acid sequence analysis shows homology to rat Ca2+/calmodulin-dependent protein kinase-II (CAB-II) and yeast phosphoprotein phosphatase (CAB-I).

• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.800-805
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• 2007

Two endoglucanases with processive cellulase activities, produced from Fomitopsis palustris grown on 2% microcrystalline cellulose(Avicel), were purified to homogeneity by anion-exchange and gel filtration column chromatography systems. SDS-PAGE analysis indicated that the molecular masses of the purified enzymes were 47 kDa and 35 kDa, respectively. The amino acid sequence analysis of the 47-kDa protein(EG47) showed a sequence similarity with fungal glycoside hydrolase family 5 endoglucanase from the white-rot fungus Phanerochaete chrysosporium. N-terminal and internal amino acid sequences of the 35-kDa protein(EG35), however, had no homology with any other glycosylhydrolases, although the enzyme had high specific activity against carboxymethyl cellulose, which is a typical substrate for endoglucanases. The initial rate of Avicel hydrolysis by EG35 was relatively fast for 48 h, and the amount of soluble reducing sugar released after 96 h was $100{\mu}g/ml$. Although EG47 also hydrolyzed Avicel, the hydrolysis rate was lower than that of EG35. Thin layer chromatography analysis of the hydrolysis products released from Avicel indicated that the main product was cellobiose, suggesting that the brown-rot fungus possesses processive EGs capable of degrading crystalline cellulose.

• Parasites, Hosts and Diseases
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• v.41 no.4
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• pp.189-196
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• 2003

In order to evaluate the possible roles of secretory proteases in the pathogenesis of amoebic keratitis, we purified and characterized a serine protease secreted by Acanthamoeba lugdunensis KA/E2, isolated from a Korean keratitis patient The ammonium sulfate-precipitated culture supernatant of the isolate was purified by sequential chromatography on CM-Sepharose, Sephacryl S-200, and mono Q-anion exchange column. The purified 33 kDa protease had a pH optimum of 8.5 and a temperature optimum of $55^{\circ}C$. Phenylmethylsulfonylfluoride and 4-(2-Aminoethyl)-benzenesulfonyl-fluoride, both serine protease specific inhibitors, inhibited almost completely the activity of the 33 kDa protease whereas other classes of inhibitors did not affect its activity. The 33 kDa enzyme degraded various extracellular matrix proteins and serum proteins. Our results strongly suggest that the 33 kDa serine protease secreted from this keratopathogenic Acanthamoeba play important roles in the pathogenesis of amoebic keratitis, such as in corneal tissue invasion, immune evasion and nutrient uptake.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.5
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• pp.908-916
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• 2000

This study was undertaken to purify the anticoagulant polysaccharide from Pachymeniopsis elliptica. The anticoagulant compound (PE-2IVc-2-1) was isolated from the crude polysaccharide (PE-2) by the sequential steps such as ultrafiltration (PE-2IV), anion-exchange chromatography of DEAE-Toyopearl 650C(PE-2IVc), size-exclusion chromatography of Sepharose CL-6B (PE-2IVc), and HPLC Shodex OHpak column chromatography (PE-2IVc-2-1). The apparent molecular weight of purified polysaccharide (PE-2IVc-2-1) was approximately 710 kDa. It was mainly composed of galactose (48.6%) and 3,6-anhydrogalactose (44.0%) with 27.1% of sulfate residue, in addition a trace amount of xylose (3.6%), glucose (1.1%), mannose (1.1%), arabinose (0.8%) and fucose (0.8%). The IR spectrum of the polysaccharide showed the characteristic bands of S=O(1116.74 and 1143.17 $cm^{-1}$) and C-O-S (828.19 $cm^{-1}$) stretching, respectively. The anticoagulant effect of PE-2 in ex vivo was proportional to the concentration of intravenously injected dose, up to 100 mg/kg.

• Journal of the Korean Chemical Society
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• v.28 no.1
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• pp.47-53
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• 1984

The separation of the rare earth elements with diethylene triamine N, N, N', N', N"-pentaacetic acid (DTPA) as eluent was carried out at different pH and concentrations by using anion exchange resin column. The rare earth elements were absorbed on the upper of the resin column and the best condition of the separation behavior was 0.025M of DTPA at pH 8.35. The elution order of the rare earths was in the order of the atomic number of the rare earth elements except samarium. The resolution of adjacent rare earth elements that have been separated with 0.025M-DTPA as eluent, was from 3.03 to 1.25 at pH 8.35. Resolution of Ce-Pr was maximum value in 3.03 and Eu-Gd was minimum in 1.25 at condition mentioned above, respectively. The resolution of rare earth elements separated with 0.025M DTPA eluent was very good at pH range of 8.0~8.6.

• Food Science of Animal Resources
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• v.21 no.4
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• pp.367-373
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• 2001

Glycoproteins PAS-6(50 kDa) and -7(47 kDa) from the bovine milk fat globule membrane share a common protein core but differ in their carbohydrate moiety. We have analyzed and proposed the structures of the N-linked sugar chains of PAS-7 by Oxford Glyco System(OGS) RAAM2000. The N-linked sugar chains were liberated from PAS-7 by hydrazinolysis and, after modifying the reducing ends with 2-aminobenzamide(2-AB), were separated into one neutral(7N, 55%) and two acidic(7M, mono-, 43%; 7D, di-, 2%) sugar chain groups. 7N was finally separated into 5 chains(a, b, c, d, and e), respectively. The structure of this 2AB-neutral sugar chain was determined by sugar analysis, exoglycosidase digestion with OGS glycosidase Kit and OGS RAAM2000 system. The results show that fraction e was the same of reported 7N1A, the biantennary complex type with a fucose on reducing end and two N-acetyllactosamine branch on non-reducing end. Therefore, it was proved that OGS RAAM2000 method is in conformity with conventional analysis of sugar chain structure from bovine PAS-7.

• Journal of the Korean Chemical Society
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• v.12 no.2
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• pp.47-50
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• 1968

Two methods for separation of the impurities from bismuth metal have been found by the use of Dowex $1{\times}4,$ anion exchange resins. The first method is that Dowex $1{\times}4$ resins are packed into the two stage columns (height of under stage is 22cm and upper stage is 3cm, and diameter of columns are 1.5cm), and the impurities of Pb(II), Ag(I) and Cu(II) are separated by the eluent of 7.5M HCl soln, Zn(II), Fe(III) by 0.5M HCl solns, and Te(IV) in the upper stage by 2M NaOH soln. Remained Au(III) in the upper stage is determined by the ignition with resins. The 2nd method is that for the separation of all the impurities simultaneously the same resins are packed into single stage column(height is 10cm and diameter is 1.5cm), and all the impurities of Pb(II), Zn(II), Cu(II), Fe(III) and Ag(I) are eluted by the eluent of 0.5M HCl soln. Separated impurities are determined by the colorimetry.

• KSBB Journal
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• v.17 no.6
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• pp.566-570
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• 2002

In lactic acid fermentation, the end product inhibition by lactic acid causes several problems. The most important of which are low lactate formation rate and its recovery from fermentation broth. To overcome these problems, extractive lactic acid fermentation was carried out in a bioreactor, which was connected to a column packed with anion exchange resin (Amberlite IRA-400, 250 g). The system was started as a batch process, and then the separation process was started when the lactic acid concentration reached 10 g/L, 20 g/L or 30 g/L. In each case, total lactic acid concentration was reached to 48.6, 53.6, 52.6 g/L with its productivity of 1.2 g/L $.$ h, 1.6 g/L $.$ h, and 1.3 g/L $.$ h, respectively Especially, in the case of the 20 g/L recycling-initiation process, extractive fermentation reduced tie fermentation time (17 hrs) by 34% in comparison with the conventional batch process. The direct consequence of this time reduction was shown by a 1.8 fold increase in overall lactic acid productivity.

• Horticultural Science & Technology
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• v.28 no.4
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• pp.593-599
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• 2010

This study was carried out to investigate the level of total glucosinolates in different parts (outer and inner part) of fresh, salted Chinese cabbage ($Brassica$ $campestris$ L. ssp. $Pekinensis$) and Kimchi at different harvesting periods (June-July, August-September, October-November, December-April, and May). For determination of total glucosinolates, Chinese cabbage was used for analytical sample preparation, provided with an anion exchanges column and measured by UV-visible Spectrophotometer. The fresh Chinese cabbage (FCC) that was harvested in June-July contained the highest level of total glucosinolates and was higher in outer part than inner part in all harvesting periods. The salted Chinese cabbage (SCC) that was harvested in May contained the lowest level of total glucosinolates. Total glucosinolates level of SCC in outer part was higher in June-July and August-September. The manufactured Kimchi (K) using harvested Chinese cabbage in June-July and August-September contained the highest level of total glucosinolates while that harvested in May contained the lowest level. The level of total glucosinolates in different parts was higher in inner part than outer part in all harvesting periods except for May. In all harvesting times, the level of total glucosinolates of FCC was higher than processed Chinese cabbage (SCC and K). Based on these results, levels of total glucosinolates are influenced by harvesting periods, parts and processing conditions of Chinese cabbage.

• Food Science and Preservation
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• v.10 no.3
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• pp.365-369
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• 2003

The main objective of this study was to investigate the pattern of glucosinolates, in terms of 3-butenyl isothiocyanate, and total glucosinolates, in different parts of young seedlings and seeds in Korean Chinese cabbages. For determination of glucosinolates, 55 days and Winter pride cultivars of Korean Chinese cabbages seeds and different parts of 1, 3, 5, and 7-day-old seedlings were used for analytical sample preparation provided with an anion exchanges column, and measured by GC and UV-visible Spectrophotometer. The concentration of 3-butenyl ITC was higher in the cotyledon and the hypocotyl parts of 55 days cultivar seedling than those of Winter Pride cultivar. In the cotyledon parts of Winter pride cultivar seedling, 3-butenyl ITC amount was increase to 3-day-old seedling and then reduced. The cotyledon of 55 days cultivar contained the highest concentration of total glucosinolates while those were increased in the hypocotyl and decreased in the root. The level of total glucosinolates was increased to 5-day-old seedling and then decreased in the cotyledon and the hypocotyl parts of seedling in Winter pride cultivar. There was no significant difference of total glucosinolates in the root part. In the seeds, 55 days cultivar concentration was higher than Winter pride cultivar both of 3-butenyl ITC and total glucosinolates. In our study, the levels of 3-butenyl ITC and total glucosinolates depended on cultivars, growth stages, and parts in Korean Chinese cabbage.