• Nutritional Sciences
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• 제9권4호
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• pp.317-322
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• 2006

Soybeans are well-known as allergenic foods. Koreans consume large amounts of soybean foods, such as kochujang, which have gone through the fermentation process. To lower the allergenicity of these foods, we prepared hypo allergenic kochujang with aloe extract (AK). A sensory evaluation was conducted along with a clinical evaluation that used a double-blind, placebo-controlled food challenge (DBPCFC) test These tests were designed to evaluate the acceptability of the fermented foods. In comparison to normal kochujang (NK), AK elicited a higher sensory test score, and the rate of positive reactions in atopic dermatitis patients during the DBPCFC test was reduced. Methods of protein extraction, protein quantitation with sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), and protein identification using two-dimensional (2D) gel electrophoresis were performed for both NK and AK to compare the functional factors. We found a reduction in the levels of high molecular proteins even though the bands of the proteins had not entirely disappeared, indicating that the boiling and fermentation process changed the soybean protein patterns. The rate of the reduction of high molecular proteins was more effective in the AK. In conclusion, AK can be recognized as a food with hypoallergenic effect.

• 대한수의학회지
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• 제5권1호
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• pp.1-16
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• 1965

I. A Comparison of Sodium Sulfate Precipitation and Zone(Paper, Agar) Electrophoresis; Many kinds of techniques have been used for fractionating serum proteins. In the present study, using bovine serum, the fractions obtained with sodium sulfate were compared with those determined by zone electrophoresis. 1. Fibrinogen was precipitated with 4 to 10 percent of sodium sulfate. 2. ${\gamma}$-globulin required 10 to 16 percent of the salt for precipitation. 3. ${\beta}$-globulin began to precipitate at 12 percent sodium sulfate, and completed precipitation at approximately 26 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. 4. ${\alpha}$-globulin completed precipitation at 13 to 28 percent sodium sulfate in paper electrophoresis and at 22 percent in agar electrophoresis. 5. Albumin began to precipitate at 14 percent of the salt, and was free from the mixture of globulins approximately at 28 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. The results of comparing fractions by the two methods were as follows: 1. Euglobulin (15%) was equal to the sum of the most ${\gamma}$-globulin and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. 2. Pseudoglobulin I (15-17.5%) corresponded to the most ${\alpha}$-, ${\beta}$-globulins and a small quantity of albumin. 3. Pseudoglobulin II(18-22%) was a mixture of the ${\alpha}$-, ${\beta}$-globulins and albumin fraction. 4. Albumin (above 22%) contained the most albumin fraction separated by zone electrophoresis and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. As mentioned above the fractions obtained with sodium sulfate were a mixture of the various proportion of the fractions determined by zone electrophoresis. The solubility of serum fractions to sodium sulfate coincided with the mobility of those by zone electrophoresis. (By percent of sodium sulfate we mean gram of sodium sulfate contained in $100m{\ell}$ of solution). II. Immunological Studies on Serum Protein Fractions with Sodium Sulfate; In the previous report the fractions of bovine serum protein with sodium sulfate compared with those obtained by zone electrophoresis, and the findings were that the former contained various proportion components of the latter. In this study the author studied whether or not the fractions with sodium sulfate are simple component antigenically by immunoelectrophoresis and micro double diffusion test (Immuno-precipitation), using rabbit antiserum to bovine serum. In immunoelectrophoresis, normal bovine serum developed with rabbit antibovine serum showed about ten distinct precipitin arcs. The distribution of these arcs was as follows: 1 albumin, 2 ${\alpha}_1$-, 3 ${\alpha}_2$-, 2 ${\beta}_1$-, ${\beta}_2$-, and 1 ${\gamma}$-globulin (Fig. 7, 9). In micro double diffusion test, five to six precipitation bands could be seen between antigens and antibody, the order of the precipitation bands location is albumin, ${\alpha}$-, ${\beta}$-, and ${\gamma}$-globulin from the side of antiserum well (Fig.19). Frequently the ${\alpha}$-, and ${\beta}$-precipitation bands were separated into two or three precipitation bands, which indicated that these globuline are not a pure component antigenically as shown in immuno-electrophoresis. In both Immunological methods, the two ${\alpha}$-, ${\beta}$-precipitin arcs and bands appeared clear and strong, indicating that the two globulins reacted as strong antigens. The precipitate reaction of ${\gamma}$-globulin was shown at 12 to 16 percent sodium sulfate; ${\beta}$-globulin at 12 to 20 percent; ${\alpha}$-globulin at 12 to 22 percent (immuno-electrophoresis), at 12 to 26 percent (Diffusion); and albumin at above 22 percent. Antigenically euglobulin contained ${\gamma}$-, ${\beta}$-, and ${\alpha}$-globulins, Pseudoglobulin I and Pseudoglobulin II were composed of ${\alpha}$-, and ${\beta}$-globulins, and albumin was a mixture of ${\alpha}$-globulin and albumin determined by zone electrophoresis. The results indicated that the fractions of serum protein obtained by either method were constituents of various proteins antigenically except ${\gamma}$-globulin and albumin by Zone electrophoresis.

• 한국식품조리과학회지
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• 제8권4호
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• pp.379-385
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• 1992

1. Amino acid compositions were determined by amino acid analyzer. Through the analysis of these samples, it was found that glutamic acid was the most abundant; glycine, aspartic acid, lysine and threonine were rich; and tryptophan and methionine were the limiting amino acid. 2. Albumins, globulins, gliadins and glutelins were extracted from the Kangwon hull, Kangwon rice buckwheat, and wheat. The relative proportions of protein fractions were 52.45 : 10.14 : 16.61 : 20.80% in Kangwon hull buckwheat, 21.10 : 13.80 : 28.40 : 36.70% in Kangwon rice buckwheat and 6.87 : 1.65 : 42.85 : 48.6% in wheat, in the order of albumins, globulins, gliadins and glutelins. 3. Polyacrylamide gel electrophoresis (PAGE) and SDS-polyacrylamide gel electrophoresis (SDS-PAGE) were performed to identify the subfractions of each protein fraction. The electrophoregrams of PAGE showed that the same fractions of both Kangwon hull buckwheat protein and Kangwon rice buckwheat protein had very similar electrophoretic patterns to each other respectively, but there were significant differences in the patterns between buckwheat proteins and wheat proteins.

• Archives of Pharmacal Research
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• 제25권5호
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• pp.704-708
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• 2002

A fast and sensitive protein staining method in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using both an acidic dye, Coomassie Brilliant Blue R-250 (CBBR) and a basic dye, Neutral Red (NR) is described. It is based on a counter ion-dye staining technique that employs oppositely charged two dyes to form an ion-pair complex. The selective binding of the free dye molecules to proteins in an acidic solution enhances the staining effect of CBBR on protein bands, and also reduces gel background. It is a rapid staining procedure, involving fixing and staining steps with short destaining that are completed in about 1 h. As the result, it showed two to fourfold increase in sensitivity comparing with CBBR staining. The stained protein bands can be visualized at the same time of staining.

• Applied Biological Chemistry
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• 제30권2호
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• pp.163-168
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• 1987

전보에서 보고한 Bradyrhizobium japonicum 9 균주와 Rhizobium fredii 7 균주의 균체 중의 단백질 Pattern의 차이를 일차원 및 이차원 전기영동법에 의하여 조사하였다. 일차원 전기영동법 (SDS-PAGE)에 의해서는 두 group의 균체에서 모두 52개의 band가 관찰되었고 그 중 6개의 main band 로써 group 간의 차이가 확인되었으며, 이차원 전기영동법(2D-PAGE)에 의한 두 group간에 단백질 구성은 Rhizobium fredii 에서는 단백질이 산성 쪽에, Bradyrhizobium japonicum 는 alkali성 쪽에 비교적 많이 분포되어 있었다. 또한 두 group 간의 균체 아미노산 조성을 조사한 결과 조선상의 뚜렷한 차이가 없었다. 분리된 근류균을 확인하는데 전기영동법이 유용하였으며, 일차원 전기영동법은 많은 균주를 신속하게 확인할 수 있었고 이차원 전기영동법은 해상력 및 분리된 단백질 spot를 분석하는데 용이하였다.

• 한국작물학회지
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• 제30권4호
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• pp.405-411
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• 1985

본 실험은 보리품종의 구분에 가장 적합한 전기영동법과 효소를 구명하고자 6개 보리품종(알찬보리, 수원 216호, 조풍보리, 사천 6호, 향맥, 히노데하다가)의 종실 단백질을 7.5％ polyacrylamide slab gel, 2-30％ polyacrylamide porosity gradient tube gel, isoelectricfocusing(pH 4-9)과 starch gel을 사용 전기영동 후 protein band pattern과 esterase, acid phosphatase, malate dehydrogenase, glutamate dehydrogenase 및 leucine aminopeptidase의 동위효소 pattern을 관찰하였던 결과는 다음과 같았다. 1. 7.5％ polyacrylamide slab gel에서 단백질 pattern과 2-30％ polyacrylamide porosity gradient tube gel에서의 단백질과 esterase의 band pattern이 품종간 뚜렷한 차이를 보여 보리품종구분에 가장 적합하였다. 2. Acid phosphatase, malate dehydrogenase, glutamate dehydrogenase 및 leucine aminopeptidase 등의 동위효소 pattern은 모든 실험한 품종이 동일한 형태를 보여 품종구분에 이용하기에는 적합하지 않았다.

• BMB Reports
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• 제39권6호
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• pp.717-721
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• 2006

In vitro analyses of type I signal peptidase activities require protein precursors as substrates. Usually, these pre-proteins are expressed in vitro and cleavage of the signal sequence is followed by SDS polyacrylamide gel electrophoresis coupled with autoradiography. Radioactive amino acids have to be incorporated in the expressed protein, since the amount of the in vitro expressed protein is usually very low and processing of the signal peptide cannot be followed by SDS polyacrylamide gel electrophoresis alone. Here we describe a rapid and simple method to express large amounts of a protein precursor in E. coli. We have analyzed the effect of ionophors as well as of azide on the accumulation of expressed protein precursors. Azide blocks the function of SecA and the ionophors dissipate the electrochemical gradient across the cytoplasmic membrane of E. coli. Addition of azide ions resulted in the formation of inclusion bodies, highly enriched with pre-apo-plastocyanine. Plastocyanine is a soluble copper protein, which can be found in the periplasmic space of cyanobacteria as well as in the thylakoid lumen of cyanobacteria and chloroplasts, and the pre-protein contains a cleavable signal sequence at its N-terminus. After purification of cyanobacterial pre-apo-plastocyanine, its signal sequence can be cleaved off by the E. coli signal peptidase, and protein processing was followed on Coomassie stained SDS polyacrylamide gels. We are optimistic that the presented method can be further developed and applied.

• BMB Reports
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• 제28권6호
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• pp.504-508
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• 1995

The electrophoretic patterns of plasma membrane surface proteins of normal rat liver cells and rat hepatomas were compared in 10% non-denaturing and 7-15% gradient non-denaturing gel. Chemical carcinogens, 2-Me DAB (2-methyl-4-dimethylaminoazobenzene) and DENA (diethylnitrosamine), were used to induce hepatoma in rats. One protein which disappeared in hepatoma was identified in normal rat liver by non-denaturing gel electrophoresis. Rabbit antisera were raised against this specific protein, and the protein was purified by Sephacryl S-200 column and immunoaffinity chromatography using the purified antibody. The purified protein showed two bands of molecular weights approximately 50 $kD_{\alpha}$ and 52 $kD_{\alpha}$ by SDS-polyacrylamide gel electrophoresis, which reacted specifically with the antibody. However only one band was observed in non-denaturing gel and also in isoelectric focusing with a pI value of 6.6. This study showed the existence of an unique protein on the plasma membrane surface of normal rat liver cells which disappeared in rat hepatomas induced by chemical carcinogens.

• 한국식품과학회지
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• 제17권1호
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• pp.1-4
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• 1985

연근별로 인삼 단백질의 생화학적 특성을 체계적으로 규명하기 위하여 polyacrylamide gel 전기영동, SDS-PAGE, gel filteration 및 아미노산 자동분석기에 의한 실험결과는 다음과 같다. 인삼의 수용성 단백질을 Sephadex G-200으로 분획하여 얻은 2개의 peak 중주 단백질은 SDS-polyacrylamide gel 전기영동에 의해 분자량은 43,000으로 추정되었다. Polyacrylamide gel 전기영동 및 SDS-PAGE에 의한 band의 수는 연근에 따라 별 차이 없이 각각 8 및 $7{\sim}11$개 확인되었다. 인삼의 수용성 단백질과 그의 주 단백질을 아미노산 분석한 결과 arginine이 각각 45.17과 57.36%로 최대의 함량을 나타내었고 methionine과 cystine의 함량은 매우 낮았다.

• 약학회지
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• 제22권1호
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• pp.42-50
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• 1978

The plasma proteins of fresh water fishes have been fractionated by disc electrophoresis in acrylamide gels utilized as an electrophoretic supporting medium. The species of fishes examined in this experiment were Anguilla japonica, Misgurnus mizolepis, Parasilurus asotus, Siniperca scherzeri, Pelteobagrus fulvidra, Carassius carassius, Cyprinus carpio, and Hemibarbus labeo, obtained in the Han River. Disc electrophoresis was performed as described by Ornstein and Davis. Gels and buffer solution were prepared by the method developed by W.J.Kim. The separation gels were 7% acrylamide gel. The fractionation of plasma proteins showed 13 bands in Anguilla japonica, 10 in Misgurnus mizolepis, 15 in Parasilurus asotus, 12 in Siniperca scherzeri, 11 in Pelteobagrus fulvidra, 13 in Carassius carassius, 9 in Cyprinus carpio, and 13 in Hemibarbus labeo. The patterns of plasma protein on the each species of fishes were different in the number of bands, ratio of contents, relative mobilities, and forms of fractionation.