• Microbiology and Biotechnology Letters
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• v.17 no.1
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• pp.19-23
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• 1989

A sequential system composed of hydroxylapatite chromatography and gel permeation chromatography was developed to purify the IgM type monoclonal antibody against the colon cancer cell SC-1 from the ascitic fluid of mice injected with the murine hybridoma CH07E02. In the hydroxylapatite chromatographic step the band dilution could be reduced by controlling the gradient and flow rate of the eluent, the sodium phospate buffer, the optimum values for these variables being 5.82$\times$10$^{-3}$M/cm and 0.2$m\ell/\textrm{cm}^2$/min, respectively. A degree of purity better than 99.99% as judged from silverstaining of the SDS-PAGE bands, was obtained by adding the gel permeation chromatographic step in tandem.

• Korean Journal of Food Science and Technology
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• v.17 no.4
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• pp.276-282
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• 1985

To characterize bitter peptides in fermented protein foods, peptides were extracted with 2:1 (v/v) chloroform-methand from various samples and separated into fractions I, II, and III by Sephadex G-25 gel chromatography. Amino acid compositions of Mozzarella cheese, soybean paste, and each fraction from the two samples were analyzed to calculate the average hydrophobicity. All the solvent extracts of the food samples had strong bitter taste, although the original samples did not taste bitter. The yield of solvent extraction ranged from 0.08 to 62.50% of total nigrogen of food samples. The average hydrophobicity calculated from the amino acid composition of Mozzarella cheese was 1376 cal/mole, solvent extract 1,623 cal/mole, gel chromatography traction I, 1,797 cal/mole, fraction II, 2,454 cal/mole, and fraction III, 1,559 cal/mole. In the case of soybean paste, the average hydrophobicity of original sample, solvent extract, gel chromatography fraction I, II, and III wre 1,229, 1,654, 1,900,998 cal/mole, respectively. The important amino acids in bitter peptides were leucine, 2016, phenylalanine, proline, and voline.

• Applied Biological Chemistry
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• v.40 no.1
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• pp.39-42
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• 1997

Angiotensin converting enzyme (ACE) inhibitors were isolated and purified from pig blood plasma. Pig blood plasma was obtained after removing blood cells by centrfugation, followed by the addition of anticoagulant to whole pig blood. To precipitate plasma proteins, pig blood plasma was treated with 4% trichloroacetic acid (TCA) as a final concentration. ACE inhibitors were isolated from plasma protein hydrolysates and TCA supernatant, using ultrafiltration, gel permeation chromatography, and reverse-phase high pressure liquid chromatography. ACE inhibitors isolated from plasma hydrolysates and TCA supernatant had $IC_{50}$ values of $23\;{\mu}M$ and $2\;{\mu}M$, pentapeptide, respectively.

• Korean Journal of Food Science and Technology
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• v.21 no.5
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• pp.606-612
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• 1989

A New lectin from baby clam, Tapes japonica, was isolated and purified through the following procedures; acetone powder, 0.15M NaCl extraction, ammonium sulfate fractionation, N-acetyl-D-galactosamine-agarose affinity column, and ion exchange Mono Q of FPLC. This lectin nonspecifically agglutinated human erythrocytes but didn't agglutinate mouse and rabbit erythrocytes. And the lectin neither stimulated human lymphocytes nor agglutinated Sarcoma 180 cells. On polyacrylamide gel electrophoresis, the lectin migrated as a major single band indicating homogeneous. A molecular weight was estimated to be about 131,000 daltons by Biogel P-300 and 125,000 daltons by SDS-PAGE without ${\beta}-mercaptoethanol$. This lectin is supposed to be a tetramer composed of heterogeneous subunits, about 30,000 and 33,000 daltons. Baby clam lectin was inhibited by EDTA and recovered agglutinating activity by $Ca^{++}\;and\;Mn^{++}$. This lectin is revealed as glycoprotein that contained about 4.2% neutral sugar.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.23 no.1
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• pp.12-24
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• 1990

Two trypsin-like enzymes, designated trypsin A and 3, purified from the intestine of menhaden by $(NH_4)_2SO_4$ fractionation, Benzamidine-Sepharose 6B affinity chromatography, DEAE-Sephacel ion exchange chromatography and Sephadex G-75 gel filtration chromatography. The two trypsins were subjected to compare the enzymatic properties of the trypsin-like enzymes from the other dark fleshed fishes. Both trypsins catalysed the hydrolysis of N$\alpha$-benzoyl-DL-arginine-p-nitroanilide and they were remarkably inhibited by several well known trypsin-inhibitors, tosyllysyl chloromethyl ketone, soybean trypsin inhibitor, be-nzamidine, leupeptin and antipain, etc. Therefore, it was ascertained that the two enzymes are serine-type trypsins. The molecular weights of these enzymes were about 25,000 and 26,200, respectively, ;Is determined by SDS-PAG electrophoresis and by Sephadex G-100 gel filtration, and the molecular weights of these two enzymes are somewhat fewer than those from the other dark fleshed fishes. Both enzymes had less basic amino acids such as arginine and Iysine, whereas they had slightly high contents of neutral amino acids, glycine, alanine and tryptophane. The enzymes showed a pH optimum of $8\~11$ at $60^{\circ}C$ against the $N\alpha$-benzoyl-DL-argi-nine-p-nitroanilide substrate and they were quite unstable above $40^{\circ}C$ and under the atidic pH region. The Km constant of the two enzymes against the $N\alpha$-benzoyl-DL-arginine-p-nitroanilide was $1.4\times10^{-4}M$ for trypsin A and $4.3\times10^{-5}M$ for trypsin B, respectively.

• Journal of the Korean Society of Clothing and Textiles
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• v.22 no.4
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• pp.477-481
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• 1998

Colored components in black tea were extracted, freeze-dried, and analysed to investigate the possibility using as a natural dye. Fractionation of the colored components was carried out by gel permeation chromatography. The colored components in black tea were elected into seven fractions. Each fraction was analyzed by UV spectrophotometer. The early fluted fractions 1-4 did not show any absorption peaks in 320-700 nm and showed the increase in absorption as it approaches to short wavelength and are considered as highly polymerized colored substances. Fractions 5-6 showed tar at 350 m and are considered as thearubigins. Fraction 7 showed absorption peaks at 376 nm and 456 nm and is considered as theaflavin. IR spectra of each fraction show: Strong C=0 stretching band at 1650 cm-1 appears in fractions 1-4, but not in fractions 5-7. Strong C=0 stretching band at 1700 cm-L appears in fraction 3-7. C=0 stretching band at 1610 cm-1 appears as a shoulder in fraction 4 and progressively changes into strong peak in fraction 5-7. From these results, it is assumed that colored components in black tea consist of polyphenolic substances having different molecular weight which were formed during tea manufacturing process. The colorants from black tea infusion were applied to silk, wool, cotton and nylon fabrics. Black tea colorants showed high affinity to wool, silk and nylon, but very low affinity to cotton fabrics.

• Korean Journal of Microbiology
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• v.49 no.2
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• pp.179-183
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• 2013

Marine-derived microbes have yielded a variety of metabolites so far. In the course of the project to find metabolites from marine microbes, an isolate of Aspergillus protuberus (SF 5767) was selected for chemical investigation. A large scale culture of this strain in PDA media was extracted with an organic solvent and the extract was fractionated by silica gel column chromatography. Repeated reverse phase HPLC of the fractions led to the isolation of three metabolites. Their chemical structures were elucidated as deoxybrevianamide E (1), brevianamide V (2), and ergosterol peroxide (3) on the basis of spectroscopic data including MS, NMR, and UV. To the best of our knowledge, chemical investigation of A. protuberus was conducted for the first time in this study.

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.269-273
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• 2005

Toll-like receptors (TLR) are important components of innate immunity in the defense against pathogens. TLRs recognize pathogen-associated common molecular patterns. TLRs are similar to the receptors involved in defense responses in plants. TLR protein is a type 1 membrane protein, consisting of an extracellular domain containing leucine-rich repeats and a cytoplasmic domain. The cytoplasmic domain delivers ligand recognition signals that result in production of anti-microbial agents. The cytoplasmic domain (amino acid 858-1032) of toll-like receptor 9 has been expressed using methylotrophic yeast Pichia pastoris. The protein expression was confirmed by Western-blot, N-terminal sequencing and MALDl-TOF mass spectrometry. The proteins have been purified by nickel affinity, cation exchange and gel-filtration chromatography.

• The Korean Journal of Mycology
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• v.23 no.4 s.75
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• pp.332-339
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• 1995

Water-soluble polysaccharide (FCW) was extracted from the fruiting body of Fomitella fraxinea with neutral sodium chloride solution. The polysaccharide was further fractionated into FCW-I and FCW-II by ion exchange chromatography. The FCW-I and FCW-II were then purified by gel permeation chromatography and named as FCW-Ia and FCW-IIa, respectively. FCW-IIa showed relatively strong immuno-stimulating activity but FCW-Ia did not. By analyses of HPLC and GPC, FCW-Ia and FCW-IIa were identified to be homogeneous and their molecular weights were estimated to be about 15,000 and 8,700, respectively. FCW-Ia consisted of fucose, galactose, and mannose as main sugars and their molar ratio was 19.5 : 63.2 : 25.0. Protein was not detected in FCW-Ia. However, FCW-IIa was composed of glucose, galactose, and mannose at a molar ratio of 1.0 : 0.3 : 0.4 and contained 0.4% protein with a higher amount of glutamic acid. A small amount of uronic acid was detected in both FCW-Ia and FCW-IIa.

• Korean journal of food and cookery science
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• v.9 no.4
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• pp.261-265
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• 1993

For the purpose of supplying the information to improve the acceptability of soy paste as the condi-ment, the changes of peptide were determined. The results were as follows; Average peptide length were decreased. It was 102 at 0 day, 15 at 10 day and 4.1 at 180 day. Peptide fraction were the same as in 60 day and 180 day. Low molecular weight peptide were not changed greatly during fermention. Peptide identified in 180 day fermentation were Ala-Ser, Gly-Glu, Glu-Ser, Asp-Glu, Asp- Tyr, Asp-Ala-Ser, Ala-Ser-Glu, Glu-Ser-Ala, and Ala-Lys-Met. In the characteristics of bitter peptide in 180 day fermentation, soy paste itself didn't show bitter taste', solvent extration fraction I'showed bitter taste. After gel chromatography, fraction I, fraction II and fraction III were obtained and fraction II were bitter peptide of low molecular weight. After gel chromatography', solvent extration fraction 2'(water extration) were divided into fraction IV, V, VI,VII and VIII. Fraction IV, V and VI showed bitter taste. Amino acids composition of the fractions showing bitter taste were like that; fr. 1: Glu- (Asp, Pro, Val, lie or Leu)-Met fr. II Pro-(Glu, Val, Phe)-lle or Leu fr. IV: Glu-(Asp, Ala, Tyr, Leu of lie)-Phe fr. V: Ala-(Met, Glu, Pro)-lle or Leu fr. VI: Asp-(Phe, Ser, fly)-Val.