• 한국축산식품학회지
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• 제18권4호
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• pp.316-321
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• 1998

In this study, we carried out to isolate lactoferrin from Korean native cattle and Holstein cow by batch extraction, ion exchange chromatography, gel filtration, and affinity chromatography. The purity of the isolated lactoferrin was higher than that of lactoferrin purchased from Sigma, when determined by SDS-PAGE and HPLC analysis. Antibacterial activity of E. coli O111 by Korean native cattle lactoferrin was lower than that of Holstein lactoferrin. A minimal inhibitory concentration(MIC) of Korean native cattle lactoferrin and Holstein lactoferrin was 2.75 mg/ ml and 1.5 mg/ml respectively. The lactoferrin hydrolyzate of Korean native cattle exhibited antimicrobial activity at 0.25 mg/ml, whereas that of Holstein cow exhibited antimicrobial activity at 0.12 mg/ml. The antibacterial potency of the hydrolyzate was at least tenfold greater than that of undigeated lactoferrin with strains tested. The effect of hydrolyzate was bactericidal as indicated by rapid loss of viability of E. coli O111.

• 한국동물학회지
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• 제35권4호
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• pp.466-473
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• 1992

Storage protein-1 (SP-1) of Gallerio mellonella was identified in hemolvmph and fat body by electrophoresis. SP-1 was purified from hemolvmph by KBr density gradient ultracentrifugation , DEAE-cellulose (DE52) ion-exchange chromatography, and gel permeation chromatography (Sephadex G-200). Purity of SP-1 was confirmed by Non-SDS PAGE and electron microscope. SP-1 is 9.4 nm in diameter and regular octahedron in shape. SP-1 has isoelectric point of 5.7 and native molecular weight of 365 K dalton and is composed of one type of subunit with molecular weight of 82 K dalton. Ttiacylslvcerol and phospholipid were found to be maior lipid components in SP-1.

• Animal cells and systems
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• 제2권3호
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• pp.367-370
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• 1998

Two molecular species of apolipophorin-III (spoLp-III) were purified from the last instar larval hemolymph of Galleria mellonella by gel permeation chromatography (Sephadex G-100), ion exchange chromatography (DE-52), heat treatment (90C for 30 min) and Mono S FPLC, and were named apoLp-III-a and apoLp-lll-b, respectively. They were indistinguishable by SDS-PAGE but could be separated by native PAGE. The molecular mass of apoLp-III determined by SDS-PAGE was approximately 18 kDa. The N-terminal amino acid sequence of apoLp-III-b revealed high similarities with the apoLp-III from Manduca sexta.

• 한국미생물·생명공학회지
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• 제23권4호
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• pp.411-415
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• 1995

The dextranase (EC 3.2.1.11) produced by Aspergillus ustus GR-98 was purified by the following sequential methods; salting-out and dialysis, gel filtration on BIO-GEL P-100, ion exchange chromatography on DEAH-cellulose, affinity chromatography on hydroxyapatite, and preparative electrophoresis. Three active fractions, dextranases 1, 11 and 111, were isolated in electrophoretically pure states, and specific activities of the dextranases were 1,276, 1,154 and 1,125 units/mg, the degrees of yield were 9.0, 3.6 and 2.2%, having 145, 131.1 and 127.8 times as those of culture filtrate in degree of purification, respectively. The enzyme purity was confirmed by the PAGE, SDS-PAGE and get permeation-HPLC.

• Environmental Analysis Health and Toxicology
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• 제12권3_4호
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• pp.15-22
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• 1997

This experiment was performed to examine the immuno-reactive characteristics of fish metal-binding protein, metallothionein (MT), and gain the practical understandings for the proposed use of fish MT as a biomarker. Liver MT induced by Cd in the silver carp was seperated and purified by gel filtration chromatography and ion exchange chromatography. The immuno-reactivity of fish MT was examined with 3 rabbit antisera. Fish MT showed little reactivity with rabbit anti-rat MT antiserum and a weak reactivity with anti-MT peptide antiserum while showed a strong reactivity with rabbit anti-fish MT antiserum. The time-course change of liver MT in the silver carp, after waterborne exposure to 1 ppm of Cd, was checked by Cd-hem method and established competitive ELISA. In both cases, the induction of liver MT showed a good increasing relationship with the exposure days. The results indicate that the fish MT can be developed as a useful biomonitoring means in the toxicological study and for the evaluation of water pollution.

• Environmental Analysis Health and Toxicology
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• 제11권1_2호
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• pp.11-17
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• 1996

The purpose of this experiment is to estabilish the sensitive and specific ELISA (enzyme linked immunosorbent assay) system for the detection of fish metallothionein (MT). Silver carp were injected with CA of 1-8mg/kg body wt. 4 times during 10 days. Silver carp was very tolerant species to CA. Cd induced MT in liver was seperated and purified by gel filtration chromatography and ion exchange chromatography and identified by spectrophotometry, native gel electrophoresis and western blot analysis. The rabbit antiserum was produced by immunizing rabbit with lyophilized MT, and the competitive ELISA system was estabilished for the detection of fish MT. In the present ELISA system, the detection limit was about 33 ng/ml. When this ELISA system was employed to determine the MT level in the supernatant sample of fish liver homogenate, the reaction curve showed a good parallel corelationship with the calibration curve over a certain dilution range. The results indicate that the competitive ELISA can be a useful tool for the detection of fish MT in the toxicological study and the evaluation of water pollution.

• 약학회지
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• 제39권3호
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• pp.306-313
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• 1995

The aim of the present study was to produce low molecular weight cathepsin B inhibitor. A strain of Streptomyces aburabiensis isolated from soil in Korea was selected and the optimum condition for the production of the inhibitor was evaluated. Glucose and soytone were selected as best carbon and nitrogen sources, respectively. From the kinetic analysis in batch fermentation, it was found that the specific cathepsin B inhibitor production rate (q$_{p}$) was linearly related to specific growh rate ($\mu$). The inhibitor in culture filtrate was purified by adsorption on activated charcoal, butanot extraction, silica gel chromatography, ion exchange chromatography using Dowex-1 (Cl form) and Amberlite IRC-50 (H$^{+}$ form), and preparative TLC. From the UV, IR, Mass spectroscopy and $^{1}$H-NMR, the inhibitor was thought to be a new inhibitor of which molecular weight was 199.

• Preventive Nutrition and Food Science
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• 제8권2호
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• pp.154-157
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• 2003

A peptide inhibiting HIV-1 pretense was isolated from the hydrolysate of manila clam (Ruditapes philippinarum) proteins digested with thermolysin. The peptide was purified by using membrane filtration, gel permeation chromatography, ion exchange chromatography, and reverse phase HPLC, The amino acid sequence of the peptide was determined to be Ile-Tyr-Glu-Gly. This tetrapeptide sequence exists in some proteins of Physarum polycephalum and Mycobacterium smegmatis. Chemically synthesized Ile-Tyr-Glu-Gly showed the $IC_{50}$/ value of 22.3 $\mu$M.

• Animal cells and systems
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• 제3권1호
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• pp.47-52
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• 1999

Change of proteins was confirmed during low temperature acclimation of overwintering larva, and some biochemical characteristics of the induced antifreeze protein-1 (AFP-1) were investigated in Protaetia brevitarsis. As the freezing point depression by the action of induced AFPs, a considerable thermal hysteresis was observed in the haemolymph and in partially purified proteins. AFP-1 was purified from the cold acclimation larvae by ammonium sulfate precipitation ion exchange chromatography, gel permeation chromatography, and electroelution. The purified AFP-1 was determined to be a glycoprotein (approximately 320 kDa, pl 5.8) composed of a single type of subunit (80 kDa). The high contents of hydrophilic amino acids (Asp, Glu, Lys, Asn, Gln, Arg, Ser, Thr) were also confirmed, showing similarity with antifreeze proteins from other insects.

• Journal of Plant Biology
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• 제12권1호
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• pp.1-6
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• 1969

General components, free sugars, and free amino acids about Zostera marina which is much distributing and producing along the coastlines of Korea have been separated and identified through Ion exchange resin and Paper chromatography method. So distrigushed are sugars contained that this was quantitated by Bertrand method. 1) The result of the general component analysis of Zostera marina shows that there are much free sugars. 2) According to Bertrand method sugar containment quantitated from terrestrial stem and subterranean stem on Po-hang and Ko-hung areas is much more in terrestrial stem than subterranean stem. 3) Seven kinds of sugars are used in experiments(Table 2) among there glucose, fructose and sucrose which keep the sweetness are very much contained. 4) Shown in Fig. 3 ten kinds of free amino acids inspected from experimental liquid by Paper chromatography are separated, which are known, and one spot which in unknown is additionally separated, and in which four kinds of essential amino acid to nutrient are contained.