• KOREAN JOURNAL OF CROP SCIENCE
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• v.39 no.4
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• pp.348-352
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• 1994

Soybean seed consists of two major storage protein, the 7S and 11S globulins. For improving the quality of soybean seed protein, an increase of 11S/7S ratio would be a desirable objective because the 11S globulin contains much more the sulfur-containing amino acids than the 7S globulin. In this study, some soybean varieties were used to investigate the analyzing method for 7S and 11S globulins. 7S and 11S globulins couble be fractionated by their different solubilities in tris buffers. Adjusting the pH and tris concentration were major factors affecting the precipitation of the two globulins. And it was possible to screen the soybean genotypes having aberrant subunit compositions of the two globulins by an sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of total soybean proteins. The ratio of 11S to 7S globulin ranged from 1.29 to 1.38. This paper also dealed with the contribution of protein components in soybean seeds to the physical properties of soycurd. It indicated that the soycurd from crude 11S was remarkably harder than that from crude 7S, and springiness and cohesiveness were slightly higher in soycurd having higher proportion of 11S. So, it may concluded that proportion of protein components in soybean seed can be important factor which controls the suitability for soycurd or other foods.

• Microbiology and Biotechnology Letters
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• v.44 no.2
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• pp.130-139
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• 2016

A bacterial strain, producing an excellent lipolytic enzyme, was isolated from the intestinal tracts of an earthworm (Eisenia fetida). The strain was identified as Aeromonas hydrophila by phenotypic, chemotaxonomic characteristics and 16S ribosomal DNA analysis, and was designated as Aeromona hydrophila PL43. The lipolytic enzyme from A. hydrophila PL43 was purified via 35−45% ammonium sulfate precipitation, DEAE-sepharose fast flow ion-exchange, and sephacryl S-300HR gel filtration chromatography. The yield of the purified enzyme was 3.7% and 2.5% of the total activity of crude extracts with p-nitrophenyl butyrate (pNPB) and p-nitrophenyl palmitate (pNPP) as substrates, respectively. The molecular weight of the purified enzyme was approximately 74 kDa using gel filtration, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and zymography. The optimal activity of purified enzyme was observed at 50℃ and pH 8.0 using pNPB, and 60℃ and pH 8.0 using pNPP. The purified enzyme was stable in the ranges 20− 60℃ and pH 7.0−10.0. The activity of purified enzyme was inhibited by PMSF, pepstatin A, Co²⁺, Cu²⁺, and Fe²⁺, but was recovered by metal chelating of EDTA. The K_m and V_max values of the purified enzyme were 1.07 mM and 7.27 mM/min using pNPB and 1.43 mM and 2.72 mM/min using pNPP, respectively.

• BMB Reports
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• v.39 no.2
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• pp.216-222
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• 2006

In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris base, 65 mM DTT(dithiothreitol) and 0.2% carrier ampholytes; (b) 5M urea, 2M thiourea, 2% CHAPS, 2% SB 3-10 (N-decyl-N, N-dimethyl-3-ammonio-1-propanesulfonate), 40mM Tris base, 65 mM DTT and 0.2% carrier ampholytes or (c) 7M urea, 2M thiourea, 4% CHAPS, 65 mM DTT and 0.2% carrier ampholytes. The optimal protocol for isoelectric focusing (IEF) was accumulated voltage of 16,500 Vh and 0.6% DTT in the rehydration solution. In the experiments conducted for the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), best results were obtained with a doubled concentration (50 mM Tris, 384 mM glycine, 0.2% SDS) of the SDS electrophoresis buffer in the cathodic reservoir as compared to the concentration in the anodic reservoir (25 mM Tris, 192 mM glycine, 0.1% SDS). Among the five protocols tested for gel storing, success was attained when the gels were stored in plastic bags with 50% glycerol. This is the first report describing the successful solubilization and 2D-electrophoresis of proteins from human lymph node tissue and a 2-D gel storage protocol for easy gel handling before mass spectrometry (MS) analysis.

• Journal of fish pathology
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• v.5 no.1
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• pp.29-35
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• 1992

This study was carried out in order to identify the biochemical and serological characteristics of Edwardsiella tarda isolated from cultured flounder, Paralichthys olivaceus in the east and south coast of Korea. During the year of 1990 and 1991, the number of isolated E. tarda were 131 strains. To identify the biochemical characteristics of them kinds of tests were conducted. The results represented that all the strains had the same biochemical characteristics, and their biochemical characteristics were no differences among strains. A serological analysis was carried out based on agglutination test with antiserum belonging to E. tarda serotype a (E-22), b (SU-138), c (SU-100) classfied in japan. The selected 10 isolates showed agglutinin titer of 5120-2560, 40-80 and below 40 against E. tarda serotype a, b and c, respectively. Sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) profiles of cell proteins of selected 10 isolates were showed no differences in kinds and volumes of proteins among strains.

• Journal of Animal Science and Technology
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• v.44 no.5
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• pp.633-642
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• 2002

Enteric colibacillosis has economically become an important disease of young animals as a result of increasing intensification of farrowing management. The objective of this experiment is to isolate fimbrial antigen from enterotoxigenic E. coli F41, to develop specific polyclonal IgY which can effectively neutralize or reduce the proliferation of pathogens in feed or living animal system, and to apply IgY technologies to animal industry. The results obtained were as follows: The molecular weight of the purified F41 antigen was 29,500 dalton on sodium dodecyl sulfate-polyacrylamide gels. Fimbrial antigen was confirmed by the western blot method. It was observed that after immunization the level of serum antibody titer of laying hen was shown in two weeks and gradually increased. The antibody titer in egg yolk appeared two weeks after it was shown in serum antibody. The titers of egg yolk antibody were gradually increased to the maximum level of 320,000 (antigen 50${\mu}g$/$m\ell$), 450,000 (antigen 200${\mu}g$/$m\ell$), and 320,000 (antigen 600${\mu}g$/$m\ell$). According to the results of specificity test by ELISA, the anti-F41 antibodies from chicken serum and egg yolk reacted only with ETEC F41 antigen. There was no cross reaction with other ETEC strains (K88, K99, and 987P). In vitro condition, as a result of antigen binding ability of yolk antibodies, bacterial concentration was rapidly decreased to $10^5$ CFU/$m\ell$ from $10^9$ CFU/$m\ell$ when 2${\sim}$4 mg/$m\ell$ of freeze dried WSF (water soluble fraction) was added.

• Journal of Korean Society of Environmental Engineers
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• v.29 no.6
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• pp.670-677
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• 2007

The stability of CGAs(colloidal gas aphrons) prepared from non-ionic and ionic surfactants was investigated. Those surfactants were sodium dodecyl sulfate(SDS), Triton X-100, Tween 80 and Quillaja Saponin. The stability of CGAs prepared from single surfactants or mixed surfactants(two components) using a CGA generate. was investigated as functions of temperature, surfactant concentration and stirring time. Saponin among the single surfactants has shown the longest duration time(143 min) and then, Triton X-100, SDS, and Tween 80 were followed by at room temperature. In case of CGAs heated up to $70^{\circ}C$, SDS endured for 116 min but Saponin lasted for only 105 mit which was a considerable reduction of the duration time of CGAs at room temperature. For mixed surfactant pairs, stability of any one pairs stood between the two. That meant no synergic effect for surfactant blending. At the higher temperature, Saponin+Triton X-100 was disclosed to be the lowest, 53 min meanwhile Saponin+SDS was the highest at ambient temperature. The CGAs, initially about 140 ${\mu}m$ in diameter, began to grow right after the agitation to be about 190 ${\mu}m$ owing to coalescence of the bubbles and then became to collapse. When heated, CGAs including Saponin tended to be smaller while the others to be larger. In summary, we found that the stability of CGAs or the duration time was greater for single surfactants and at room temperature rather than for mixed surfactants that caused substantial intermolecular interactions in the CGA structure and at the higher temperature.

• Parasites, Hosts and Diseases
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• v.27 no.1
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• pp.1-8
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• 1989

Serodiagnosis of parasitic infections is widely used, since parasites or their eggs are not always detected by ordinary methods. The sensitive tests such as ELISA are highly dependent on the purity of antigens used. To solve this problem. many workers have tried to find species-specific components of antigens, The present study was performed to determine the antigenic profile of crude saline extracts of 3, 5, 8 and 12-week old p. westermani worms, which were collected from experimentally infected cats, based on SDS-PAGE and immunoblot technique. The results were as follows: 1. The SDS-PAGE showed at least 30 Protein bands ranging from 229 kDa to 10 kDa molecular weight. The protein components of p. wsstermani changed chronologically during its developmental period. The 229 kDa band was recognized only in 12-week old worms ($$SEP_{l2}$). 2. Analysis by ELISA showed a significant increase in antibody levels at 3 weeks in infected cats using crude saline extract antigens ($SEP_3,{\;}SEP_5,{\;}SEP_8,{\;}SEP_{l2}$). 3. By EITB using $SEP_3$ and $SEP_5$ infected cats recognisea major protein bands with molecular weight of 60, 35, 28, 25 or 21 kDa at 3~12 weeks of infection, and 3 additional antigens, 19, 13 and 10 kDa, were detected at 8~12 weeks of infections. 4. Using $SEP_8$ 5 antigens, 91, 85, 31, 25 and 21 kDa, were consistently detected by all infected sera tested. In addition, 3 antigens of lg. 13 and 10 kDa were detected at 8~12 weeks of infection. Using $SEP_12$, similar results were obtained with that by using $SEP_8$ and 1 additional antigen of 229 kDa, specifically reacting with the sera from 12 weeks of in(traction, was recognized.

• Applied Biological Chemistry
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• v.20 no.1
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• pp.26-32
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• 1977

The multiple 7S globulins composed of two fractions (A and B) in the electrophoresis with Davis' method were isolated at different stages of the soybean seed development. Electrophoresis of their subunits liberated in PAWU solvent [phenol-acetic acid-water (2 : 1 : 1) solution plus 5M urea] yielded 4 major bands. Observation of both the electrophoretic bands of the multiple 7S fractions(7S-A and 7S-B) and those of their subunits was suggestive of a similarity of the subunit pattern between two 7S fractions. The two fractions in multiple 7S globulins were isolated with DEAE-Sephadex A-50 column$(2.0{\sim}100cm)$ chromatography. They were separated into 2 fractions in a linear gradient concentration of 0.28 to 0.40M NaCl with phosphate buffer (pH 7.8) containing 10mM ${\beta}-mercaptoethanol$(ME). The isolated protein was dissociated into subunits with two different solvent systems; in PAWU solvent and in Tris-HCl buffer(pH 8.0) containing 1% sodium dodecyl sulfate (SDS) and 40mM ME. The dissociated subunits were subjected to electrophoresis in PAWU-treated 7.5% acrylamide gel and in 1% SDS-treated 5.6% acrylamide gel. In PAWU gel electrophoresis, total 7S globulin was separated into 5 major bands, two of which were occupied in common by two 7S fractions(7S-A and 7S-B). In SDS gel electrophoresis, total 7S globulin was separated into 7 major bands, three of which were overlapped with the subunit of the two 7S fractions. The above results alluded us to the presence of a common and/or similar subunit between the multiple 7S globulins.

• The Korean Journal of Zoology
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• v.33 no.1
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• pp.63-69
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• 1990

To study the follicular atretic mechanism in mammalian ovary, the medium-sized (3.0-6.0mm) follicles of porcine ovary were morphologically classified as nonnal and atretic. Steroid concentrations in the follicular fluid were analyzed by radioimmunoassay, and the proteins in that fluid were electrophoretically separated. Concentrations of progesterone in the atretic follicular fluid of follicular phases were higher than those of normal ones (p < 0.05). Concentrations of testosterone were high in normal luteal and atretic follicular-phases follicles. The concentrations of estradiol remained significanily lower in atretic follicular-phases follicles than normal. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis of follicular fluid proteins, four kinds of proteins (20K, 32K, 33K, 38K) were detected, and those proteins were not present in sera. According to the ovarian cycle, proteins of MW of 112K and 141K were identified. In atretic follicular fludies, MW of 23K and 24K were specifically detected. From the above results, we can conclude that, as ovarian cycle changes, steroid contents and protein composition in atretic follicular fluid are different from the normal developing follicular fluid. To further understand the physiologic roles of the proteins present in the atretic follicular fluids, these proteins need to be characterized and identified.

• The Korean Journal of Zoology
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• v.28 no.1
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• pp.31-43
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• 1985

The $(Ca^{2+}+Mg^{2+})$-ATPase has been purified homogeneously from sarcoplasmic reticulum of rat skeletal muscle by sucrose density gradient centrifugation. The purified enzyme has a molecular weight of 115,000 as judged by polyacrylamide gel electrophoresis in the presence of sodium dedecyl sulfate, and therefore has the same size of the enzyme in rabbit and chick skeletal muscle. $Ca^{2+}, Mg^{2+}, Fe^{2+}, Co^{2+}, and Mn^{2+}$ at 50 $\\muM$ show stimulatory effect on the ATP-ase, while $Zn^{2+}, Cu^{2+}, and Hg^{2+}$ inhibit it at the same concentration. The ATPase activity is insensitive to antimalarial drugs such as quinine and quinacrine, but is sensitive to inhibition by p-hydroxymecurie benzoate and phenylmethylsulfonylfluoride. The enzyme has optimum pH of 6 to 7 and Km value for ATP is estimated to be 98 $\\muM$. Thus, a number of biochemical properties of this enzyme appear to be different from those of the enzyme that have been isolated from rabbit skeletal muscle. The $(Ca^{2+}+Mg^{2+})$-ATPase appears to be selectively degraded in microsomal fraction. The activity of metalloendoprotease is evident in the microsomal preparation when assayed by radioactively labeled protein substrate, such as $^{3}H-casein and $^{125}I$-insulin. However, it is presently unclear whether the metalloendoprotease is responsible for the degradation of the $(Ca^{2+}+Mg^{2+})$-ATPase.