• Microbiology and Biotechnology Letters
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• v.26 no.5
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• pp.379-386
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• 1998

Deterioration of food is in general caused by the presence of microorganisms and chemical compounds of food itself. There exists antimicrobial compound in the food, however, addition of food antiseptics, additives, or physico-chemical processing is a common practice. The safety of artificial chemical antiseptics became a serious public concern, therefore, new natural antiseptic compounds are in need to be developed. We have isolated a new natural antifungal protein (KBS-B2) from Astragal seed through ammonium sulfate precipitation and column chromatography using FPLC Mono-S and Superose 12HR. The purified protein inhibited growth of Candida albicans, and spore germination of food spoiling fungi such as Aspergillus ochraceus, Penicillium expensum, P. digitatum and Botrytis cineria. Antifungal effect of the KBS-B2 protein could be directly assayed by bioautography overlaying the fungal spores on the electrophoresed acrylamide gel. The comparison of N-terminal amino acid sequences of the KBS-B2 with known antifungal protein revealed that had 50% homology to thaumatin and zeamatin like proteins.

• Current Research on Agriculture and Life Sciences
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• v.7
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• pp.109-116
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• 1989

This study was carried out to confirm the agent responsible for the antibacterial activity in milk culture or Lactobacillus bulgaricus to extract and purify it. The following results were summarized as followings : The antibacterial agent was extracted from the cultured skim milk with methanol and acetone and was purified by Sephadex G-50 gel filteration and thin layer chromatography on silica gel. The antibacterial substance other than lactic acid was confirmed by turbidimetric technique using the neutralized culture filtrate which inhibited the growth of Bacillus subtilis. The purified agent showed inhibitory activity against Bacillus subtilis, Escherichica coli, Pseudomonas fluorescens Staphylococcus aureus, Proteus vulgaris and Shigells $dysenteria^2$. The agent obtained from thin layer chromatography was free from $H_2O_2$ or lactic acid.

• Journal of Environmental Science International
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• v.5 no.1
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• pp.43-50
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• 1996

Thiobacillus neopolitanus R-10, which produces a active thiosulfate oxidase was isolated from nightsoil. The optimal culture conditions of Thiobacillus neopolitanus R-10 for the production of enzyme was determined as followed: 0.8% $Na_2S_2O_3$, 0.2% $KH_2PO_4$, 0.2% $K_2HPO_4$, 0.04% $Na_2CO_3$, 0.02% $MgSO_4$.$7H_2O$, 2ml trace elements solution, ann PH 6.5 at 3$0^{\circ}C$ and 72hr cultivation. The oxidase was successively purified 83 folds yield by ${(NH_2)}_2SO_4$ fractionation, DEAE-Cellulose, Sephadex A-50 column chromatogrophy and gel Sephadex G-150 gel filteration with yield of 5.9%. The molecular weight of purified enzyme was estimated to be 43.000 dalton by SDS-polyacrylamide gel electrophoresis and gel filteration column chromatography The enzyme activity was highest at 40t and PH 7.0 The enzyme activity was relatively high by $\beta$-mercaptoethanol but strongly inhibited by cysteine.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.20 no.4
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• pp.273-281
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• 1987

In this work the antioxidant effects of browning reaction products prepared by xylose-tryrtophan reaction system were discussed. The antioxygenic brown pigments were separated by solvent extraction, and column chromatography and isolated by gel filteration. The functional groups of the brown pigments which had antioxidant activity were examined. The brown pigments extracted with methanol showed antioxidant effect and were fractionated in 5portions on DEAE-cellulose column. The elutes with methanol: acetic acid(10:30 v/v sol n(A), methanol: chloroform(95:5 v/v) sol n(C), and chloroform: acetic acid(10:30 v/v) sol n(E) only showed antioxidant activity and their compositions were 22.43, 21.51 and $34.43\%$ respectively. When each fraction on DEAE-cellulose column was reseparated on Sephadex LH-20 column, 2 fractions were obtained from portion A and C respectively. Molecular weights of A, C and E fraction of brown pigments were from 2,600 to 3,700. By elucidation of IR spectra, the pigment fractions which showed a strong antioxidant activity were tearing the indole group. It is suggested that the antioxidant function of the brown pigment is due to hydroxy and amino group. A higher activity of the brown pigment fraction E might be attributed to carboxylic acid or carboxylic ester compounds.

• Food Science and Biotechnology
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• v.16 no.6
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• pp.873-878
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• 2007

In this study, we isolated porphyran was isolated from the red seaweed Porphyra yezoensis and assessed in terms of in vitro anti-proliferative activity. Sequential anion-exchange and gel-filtration chromatography led to purification of 3 porphyrans of different molecular masses, which contained <$50\;{\mu}g/mL$ protein and >$10\;{\mu}g/mL$ porphyran. Crude porphyran inhibited cell growth in a dose-dependent manner (0-5 mg/mL). When HT-29 colon cancer cells and AGS gastric cancer cells were cultured with various concentrations of the purified porphyran, cancer cell growth was inhibited by 50% at a low concentration (5 or $10\;{\mu}g/mL$). Furthermore, the polysaccharide portion of the porphyran preparation, rather than the protein portion, is the most effective at inhibiting cancer cell proliferation via apoptosis, as indicated by increased caspase-3 activity. Our results indicate that purified porphyran has significant in vitro anti-proliferative activity (p<0.05).

• Journal of Dairy Science and Biotechnology
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• v.17 no.1
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• pp.11-15
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• 1999

The objective of this study was to extract and purity the antibacterial agent from the fermented milk with Lactobacillus helveticus CH-1. The extraction and purification of antibacterial agent from the Lb. helveticus fermented milk were carried out by methanol extraction, acetone extraction, Sephadex G-200 gel filteration and thin layer chromatography and the results were as followings. The antibacterial activity of methanol-acetone extraction showed antibacterial activity against test organisms, B. subtilis, E. coli, Pseu. fluorescens, Sal. typhimurium, Shi. flexneri, and Sta. aureus. Sephadex G-200 gel chromatography showed only antibacterial activity from 33 to 37th fractions of 60 fractions. The agent purified from TLC plate confirmed the antibacterial activity by the means of bioautography.

• Journal of the Korean Society of Food Science and Nutrition
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• v.18 no.3
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• pp.279-286
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• 1989

The alkaline protease producing mold isolated from and identified as Aspergillus fumigatus. It was found that the production of alkaline protease reach to maximum was cultured for 3 days at $30^{\circ}C$. The enzyme was purified 86.13 fold and yield of the enzyme purification was 6.4%, The purification procedure include ammonium sulfate treatment, gelfiltration on Sephadex G-25, G-75, G-150 and DEAE-cellulose ion-exchange chromatography. When the purified enzyme was applied sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the molecular weight was estimated about 63000. This enzyme composed 17 amino acids and main amino acids of this enzyme were glycine and glutamic acid.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.5
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• pp.432-442
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• 1992

To identify the histological site of synthesis of yolk protein precursor, vitellogenin, by immunocytochemical method in the freshwater crab Eriocheir japonicus, we purified the yolk protein, vitellin, from crude egg extracts, and prepared the anti-rabbit serum against vitellin. Then, the site of vitellogenin synthesis was demonstrated by immunotytochemical method with PAP(peroxidase-antiperoxidase) reaction using the rabbit antiserum aganist vitellin. Female specific serum protein was identified in female serum by immunoelectrophoresis and Ouchterlony's immunodiffusion test for mature male and female sera. Based on the immunoelectrophoresis and Ouchterlony's diffusion test for mature male and female sera and crude egg extracts using antiserum against vitellogenic female serum absorbed with male serum, the female specific serum protein was identified as vitellogenin, detected in female serum only. The major yolk protein, vitellin, was purified from the crude egg extracts by DEAE-cellulose ion exchange chromatography, followed by sepharose CL-4B gel filteration chromatography. The molecular weight of vitellin was estimated to be about 245,000 dalton by sepharose CL-4B gel filteration chromatography. from the results of immunological analysis for vitellin, it was found that the vitellin antiserum contained the antibody against vitellogenin. In the results of immunocytochemical reaction by PAP method with the rabbit antiserum against vitellin, the vitellogenic oocytes and the hepatopancreas of mature female showed positive PAP reaction, but not in follicle cells and previtellogenic oocytes nf ovary, muscle of female and mature male hepatopancreas. Therefore, it showed that the hepatopancreas of mature female is the site of vitellogenic synthesis.

• Journal of Life Science
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• v.13 no.5
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• pp.718-722
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• 2003

Various marine microorganisms were isolated from seaweed, and their alginate-degrading activities were investigated. An alginate-degrading bacteria, Vibrio sp. AEBL-211, showed highest level of alginase activity when cultured on a mineral salt medium containing 0.7%(w/v) sodium alginate as the sole carbon source. The intracellular alginase from AEBL-211 was partially purified by ion chromatography on DE 52-cellulose column and gel filteration on Sepacryl G-200 column, and showed guluronate-specific 1yase activity.

• Korean Journal of Food Science and Technology
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• v.28 no.5
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• pp.859-864
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• 1996

The rhamnan sulfat extracted from green algae seaweed, Monostroma nitidum was characterized on sugars, sulfate compositions and molecular structure. Rhamnan sulfate was extracted with boiling water, and purified with two steps of cetylpyridinium chloride and ion exchange chromatography. The yield of crude rhamnan sulfate was about 2% from raw material. Rhamnan sulfate fraction, F-4 was composed of 30% rhamnose, 0.9% arabinose, 2.5% xylose, 2% glucose and 32.6% sulfate. Rhamnan sulfate F-4-3 obtained from F-4 fraction was composed of 36.8% rhamnose, 3.6% xylose, 2.7% glucose, 1.4% galactose and 30.8% sulfate. The molecular weight of F-4-3 fraction was estimated as 10,000-10,300 dalton with Sephacryl S-200 gel filtration chromatography.