• Korean Journal of Food Science and Technology
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• v.31 no.5
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• pp.1378-1385
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• 1999

The antioxidative activity of Fermented Anchovy on linoleic acid autooxidation was investigated in an aqueous system at pH 7.0. All solvent fractions from Fermented Anchovy were exhibited the strong antioxidative activity. Especially, BuOH and aqueous fractions were gained large amounts with strong antioxidative activity. Ultrafiltration, dialysis, heat treatment of aqueous fraction indicated that water-soluble antioxidants of Fermented Anchovy were heat-resistant, amino acid related compounds with smaller molecular weights than 1,000. Unbound fractions from DE-52 anion exchange chromatography were exhibited antioxidative activity with or without $15\;{\mu}M\;Fe^{+++}\;ion$. We were able to purify one methionine derivative from lots of antioxidative substances in Fermented Anchovy aqueous fraction by gel filtration, anion-exchange chromatography, TLC and HPLC, successfully. These data suggest that Fermented Anchovy aqueous fraction is a mixture of fermented small molecules with strong antioxidative activities.

• Applied Biological Chemistry
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• v.40 no.3
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• pp.178-183
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• 1997

A protease was purified from Bacillus subtilis CCKS-111 by ammonium sulfate treatment, DEAE-cellulose ion-exchange chromatography, Sephadex G-100 gel filtration and high performance liquid chromatography (HPLC). The specific activity of the purified enzyme was 24.3 unit/mg protein and the purification fold of enzyme was 50.6. Molecular weight of the purified enzyme estimated about 28,000 by HPLC gel filtration. The amino acid residues of this enzyme were 251.3 except threonine, serine and glycine. This result was similar to Bacillus subtilis subtilisin DY. From the first N-terminal amino acid to the 32th amino acid, the amino acid sequence was estimated after RP-HPLC elution. N-terminal and the 32th amino acids were alanine and aspartic acid. Alanine, serine, glycine and arginine were four major acids in the enzyme.

• Applied Biological Chemistry
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• v.35 no.1
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• pp.23-29
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• 1992

Bromelain was purified from Korean pineapple, Ananas comosus, L. The enzyme was purified about 21 fold by DEAF-cellulose ion-exchange chromatography and gel filtration on Sephadex G-150. Purified enzyme was confirmed as active single band by polyacrylamide electrophoresis and the molecular weight was estimated to be about 22,000 by SDS-PAGE. The optimum pH and temperature were 6.0 and $60^{\circ}C$, respectively. The range of its stability to the pH and temperature were respectively 5.0 to 7.0 and below $50^{\circ}C$. It was found that $Mn^{2+}$ increased the enzyme activity, whereas $Mg^{2+}\;and\;Fe^{2+}$ decreased it abruptly. The purified enzyme was inhibited by p-chloromercuribenzoic acid, indicating that reactive SH groups are required for the enzyme activity. The reaction of the enzyme followed typical Michaelis-Menten kinetics with Km value of $5.747{\times}10^{-4}\;M\;and\;Vmax\;of\;131.58\;{\mu}g/min$ for casein. When meat was treated with the enzyme, free soluble nitrogen and amino acid nitrogen increased as enzyme concentration increased.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.551-558
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• 1992

The extracellular endoinulase from Streptomyces sp. 556 was purified and characterized, The culture broth was fractionated by ammonium sulfate saturation followed by DEAE-cellulose column chromatography and 5ephadex G-200 gel filtration, The ultimately purified fraction revealed a single band in 7.5% polyacrylamide gel electropherogram. The purified enzyme showed the maximal activity at pH 5.5-6.0 and $50^{\circ}C$, but lost 93% of inulase activity after 30 min incubation at $55^{\circ}C$ . The essen.tial amino acid residue for catalytic activity appeared to be tryptophan. This endo inulase was activated by $Mn^{2+}$, whereas inactivated by $Ag^{+}$, $Hg^{+}$, $Cu^{2+}$, $Zn^{2+}$, $Fe^{3+}$ and $Mo^{6+}$ EDTA and 8-hydroxyquinoline inhibited the enzyme so that the enzyme was considered to be a metalloenzyme. The Km value for inulin was 0.287 mM, and no invertase or $\alpha$-glucosidase activity was found in the enzyme.

• Journal of the Korean Society of Food Science and Nutrition
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• v.7 no.2
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• pp.7-12
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• 1978

This experiment was conducted to fractionate the trypsin inhibitors and its heat stability of five varieties of soybean. It was observed that water extractable protein of all varieties used was fractionated into three peaks and the second peak seemed to show trypsin inhibitor activity. (TIA). The trypsin inhibitors were fractionated into four fractions-FI, FII, FIII and FIV. FIII showed the highest TIA in Suwon No. 81, Suwon No. 82 and Suwon No. 83 In the case of Kyungnam No. 3 and Suwon No. 62, the highest activity was found in FIV. When trypsin inbibitors fractionated in boiling water-bath for 20 min, the FIV showed the highest heat stability and FI was found to be the weakest.

• The Korean Journal of Zoology
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• v.36 no.3
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• pp.319-328
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• 1993

All vertebrates other than lampreys exhibit multiple loci encoding lactate dehydrogenase (EC 1.1.1.27,LDH). From the result shown by cellulose acetate and starch gel electrophoresis, the lampreys were-reported to have only one isozyme. However in our results the LDH of skeletal muscle, heart and kidney in Lampetra japonica were separated into three isozymes and that of liver was separated into two isozymes by polyacrylamide gel electrophoresis. The LDH of skeletal muscle and heart were separated into four isozymes and that of liver was separated into two isozymes by polyacrylamide gel isoelectric focusing (PAGlEF). The LDH of skeletal muscle were separated into four isozymes through the chromatofocusing. The molecular weight of LDH isozymes in skeletal muscle was approximately estimated to be 140,000 by Sephadex G-200 gel filtration. The LDH isozymes of skeletal muscle, heart and liver were inhibited by pyruvate to the nearly similar degree. And the degree of inhibition by pyruvate showed the value between LDH A$_4$and LDH B$_4$isozyme. And the LDH isozymes in heart, liver and skeletal muscle were thermostable. The results mentioned above indicate that the LDH isozyme in lamprey (Lampetra japonica) has not one isozyme but isozymes. And it is also found out that the two structures of their subunits are similar each others.

• KSBB Journal
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• v.10 no.5
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• pp.589-597
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• 1995

Protein-bound polysaccahrides(PBP) were isolated, purified, and characterized from the sawdust media after harvesting the fruit body of Flammulina velutipes. The yield of the crude PBP(Fr.CA) extracted from the sawdust media, was 0.367% relative to the original sawdust media. The total sugar and protein contents of Fr.CA were 19.8% and 23.8% respectively. Using the membrane filtration, the fraction of which the molecular weight is over 300 kDa(Fr.A) was isolated from the Fr.CA and the yield was 44.6% relative to the Fr.CA. This result indicates that high molecular PBP is the dominant components of the Fr.CA. The Fr.A was separated into three fractions (Fr.A-1, Fr.A-2 and Fr.A-3) whose yields are 5.8%, 8.5% and 13.2% respectively. These fractions were further purified using gel filtration, obtaining a single peak in each fraction that considered as pure PBP Among them, the yield of Fr.A-1-${\alpha}$ was 20.9% relative to the Fr.A-1, and the molecular weight was 800 kDa. Monosaccharide components such as glucose, galactose, mannose, xylose and fucose could be detected all fractions by a HPLC analysis. Especially in the Fr.A-1-${\alpha}$ fraction, the content of glucose and galactose appeared to be high.

• Korean Journal of Pharmacognosy
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• v.24 no.4
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• pp.267-281
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• 1993

To find antitumor components from higher fungi, the mycelia of Collybia confluens (Pers. ex Fr.) Kummer were cultured in artificial media. For efficient production of the mycelia, the influences of various modifications of culture conditions were examined. A water-soluble protein-bound polysaccharide fraction, Fr. A, was obtained from the mycelia by hot water extraction. When Fr. A was purified and fractionated by DEAE-cellulose and Sepbadex G-200 gel filtration chromatographies into four fractions which were designated B, C, C-I and C-II. The tumor inhibition ratios of these fractions ranged from 46% to 75% against the solid forms of sarcoma 180 in ICR mice at doses of 20 and 50 mg/kg/day when given intraperitoneally. Especially, Fr. C which was named Collyban(CB) exhibited a marked life-prolonging effect of the mice against ascitic forms of sarcoma 180 at a dose of 50 mg via i.p. administration. To extend spectra of the antitumor activities and eliminate the effects of allograft rejection, the characterization of antitumor effects of CB was performed in syngeneic host-tumor systems. It did not show any antitumor activity against L1210 murine leukemia in $CD_2Fl$ mice but prolonged their life span against ascitic forms of $MM_{46}$ carcinoma in $C_3H/He$ mice. Also it exhibited antitumor activity against human cervical cancer HeLa cells that were xenografted into nude mice having BALB/c genetic backgrounds by the i.p. injection at a dose of 100 mg/kg/day. In order to characterize the antitumor components, CB was examined by chemical analysis. It was acidic protein-bound polysaccharides composed of 31% polysaccharide, 27% protein and 3% hexosamine. CB was fractionated into two fractions, Fr. C-I(M.W.: 500 Kd) and Fr. C-II(M.W.:30 and 8 Kd) by Sephadex G-200 gel filtration chromatography.

• Parasites, Hosts and Diseases
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• v.50 no.1
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• pp.37-43
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• 2012

Although schistosomicidal drugs and other control measures exist, the advent of an efficacious vaccine remains the most potentially powerful means for controlling this disease. In this study, native fatty acid binding protein (FABP) from $Fasciola$ $gigantica$ was purified from the adult worm's crude extract by saturation with ammonium sulphate followed by separation on DEAE-Sephadex A-50 anion exchange chromatography and gel filtration using Sephacryl HR-100, respectively. CD1 mice were immunized with the purified, native $F.$ $gigantica$ FABP in Freund's adjuvant and challenged subcutaneously with 120 $Schistosoma$ $mansoni$ cercariae. Immunization of CD1 mice with $F.$ $gigantica$ FABP has induced heterologous protection against $S.$ $mansoni$, evidenced by the significant reduction in mean worm burden (72.3%), liver and intestinal egg counts (81.3% and 80.8%, respectively), and hepatic granuloma counts (42%). Also, it elicited mixed $IgG_1/IgG_{2b}$ immune responses with predominant $IgG_1$ isotype, suggesting that native $F.$ $gigantica$ FABP is mediated by a mixed Th1/Th2 response. However, it failed to induce any significant differences in the oogram pattern or in the mean granuloma diameter. This indicated that native $F.$ $gigantica$ FABP could be a promising vaccine candidate against $S.$ $mansoni$ infection.

• Microbiology and Biotechnology Letters
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• v.18 no.5
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• pp.496-500
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• 1990

When pepsin was used at a concentration of 8 mglml for hydrolysis of 0.02% casein, inhibitory activity of this inhibitor was proportional to a inhibitor concentration of 20 ${\mu}g$/ml, and fifty percent inhibition ($IC_{50}$) was observed to be 15 ${\mu}g$/ml. The inhibitor was pH-stable at pH range of 5-9 at $100^{\circ}C$ for 10 minutes and thermo-stable at pH 7.0 at $100^{\circ}C$ to give 100% activity for 20 minutes. The formation of pepsin-inhibitor complex was confirmed by sephadex 6-25 gel filtration and type of inhibition was determined as non-competitive inhibition by Lineweaver-Burk plot. The inhibitor strongly inhibited acid proteases such as pepsin and renin, and it was soluble in methanol very well. On TLC analysis of silicagel 60 using various sohent systems, the inhibitor gave a single spot at Rf range 0.4-0.6. From the result of IR spectrum and color reaction (Rydon-Smith, Biuret), this inhibitor was considered as peptide substance. Melting point and elemental contents were 220-$230^{\circ}C$, and C 50.61%-H 8.02%-N 9.34% (found), respectively.