• Title/Summary/Keyword: SDS-PAGE pattern

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Derivatization of Rice Wine Meal Using Commercial Proteases and Characterization of Its Hydrolysates (단백질가수분해효소를 이용한 탁주박의 가수분해 및 그 분해물의 특성)

  • Kim, Chang-Won;Choi, Hyuk-Joon;Han, Bok-Kyung;Yoo, Seung-Seok;Kim, Chang-Nam;Kim, Byung-Yong;Baik, Moo-Yeol
    • Korean Journal of Food Science and Technology
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    • v.43 no.6
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    • pp.729-734
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    • 2011
  • With the goal of transforming rice protein from an insoluble to a soluble form to increase the industrial utilization of rice wine meal (RWM), RWM was derivatized using commercial proteases and the RWM hydrolysates were characterized. Eight commercial proteases were used individually or in combination for hydrolysis of RWM. The degree of hydrolysis was assessed by determining the soluble protein in supernatant using the Lowry assay, protein in precipitates using a semimicro Kjeldahl procedure, and gravimetrically by the weight difference before and after hydrolysis. Protamex, Alcalase and Protease N proteases were most effective for hydrolysis of RWM. Although these assessment methodologies displayed some variation, they generally showed a similar pattern. When the aforementioned three proteases were simultaneously used to treat RWM, no significant difference was observed between the three assays (p<0.05) indicating an absence of enzymatic synergy.

Conditions of Water Activity of Raw Material and Adding Levels of Papain and Glucose for Processing Fermented Squid with Low Salt Concentrations (저염 오징어젓갈의 제조를 위한 원료어육의 수분활성도와 papain과 glucose의 첨가조건)

  • Jo, Jin-Ho;Oh, Se-Wook;Kim, Young-Myoung;Chung, Dong-Hyo
    • Korean Journal of Food Science and Technology
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    • v.30 no.1
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    • pp.62-68
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    • 1998
  • To develop methods to produce low salt fermented squid with rich flavor and acceptable shelf life, the optimum processing conditions such as water activity of raw material, amounts of NaCl, papain and gucose were investigated. Water activity of squid meat was adjusted to 0.94 (raw meat), 0.90 and 0.88 by cold air blast and each was salted with 3, 5, or 7% NaCl followed by fermenting at $10^{\circ}C$ for 6 weeks. Amino nitrogen was increased rapidly with high water activity and low NaCl concentration. As a result of organoleptic evaluation it was concluded that optimum conditions were to adjust water activity of raw material to 0.90 and to salt with 5% NaCl. When squid meat adjusting water activity to 0.90 was treated with 0.05, 0.1 and 0.5% papain and fermented at $10^{\circ}C$ for 6 weeks, SDS-PAGE pattern showed rapid breakdown of myofibrilar protein with increasing amounts of papain but the treatment with 0.1% enzyme was best organoleptically. pH values of squid meat added with 1 and 2% glucose were maintained lower than control (glucose 0%) but there were no significant differences between the two glucose treatments. Therefore, it was thought that adding of glucose might be extended shelf life of fermented squid with low salt concentration.

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Functionality of Extracted Proteins by Additives and Ionic Strength (첨가제의 종류와 이온강도에 따라 추출되는 육단백질의 기능적 특성)

  • Rhee, Min-Suk;Lee, Jun-Sup;Koh, Kyung-Chul;Kim, Young-Kyo;Kim, Byoung-Chul
    • Korean Journal of Food Science and Technology
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    • v.30 no.1
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    • pp.69-76
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    • 1998
  • This study was investigated to determine the effect of additives and ionic strength on the functionality of extracted proteins in preblends in order to use less additive in restructured meat products. Preblends contained the combinations of sodium chloride (NaCl; 0, 4.5, 9.0%), sodium tripolyphosphate (STPP; 0, 2.5, 5.0%), and tetrasodium pyrophosphate (PP; 0, 2.44, 4.88%). The pH values increased linearly with increasing STPP and PP concentrations (p<0.01). In the equivalent ionic strengths, PP was more effective than STPP in increasing pH. Phosphate ions were more effective on total extractable protein (used 1 M NaCl buffer) than chloride ion at equivalent ionic strengths. Solubility was decreased by adding NaCl and increasing total extractable proteins. Meat sulfhydryl contents were high with increasing total extractable proteins. When protein extracts were heated at $65^{\circ}C$, 7 min, meat sulfhydryl contents decreased and surface hydrophobicity increased (p<0.01). However, all protein extracts showed no differences in SDS-PAGE pattern. In conclusion, PP is more effective than STPP in order to use less additive but there was no linear relationship between functionnal improvement and ionic strength.

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Identification of the Protein Function and Comparison of the Protein Expression Patterns of Wheat Addition Lines with Wild Rye Chromosomes (야생 호밀 염색체 첨가 밀 계통의 단백질 발현 양상 비교 분석)

  • Lee, Dae Han;Cho, Kun;Woo, Sun Hee;Cho, Seong-Woo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.64 no.4
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    • pp.373-383
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    • 2019
  • The objectives of this study were to compare the protein expression patterns and degrees and identify the protein function of disomic addition lines (DAs) in Leymus racemosus, in order to improve the quality of wheat. Upon SDS-PAGE, L. racemosus showed two major protein bands whereas Chinese Spring (CS) had four major protein bands of high molecular weight. The DA(s) generally showed a similar protein expression pattern to that of CS, because 42 chromosomes were from CS and two chromosomes were from L. racemosus. However, only the L.r[J] line showed two protein bands of between 15 and 20 kDa, like L. racemosus. Image analysis based on 2-DE revealed that L.r[F] had the most upregulated protein spots, whereas L.r[N] had the least upregulated protein spots. For L.r[I], the frequency of the downregulated protein spots was higher than that of the upregulated ones. Using MALDI-TOF MS, the protein function was identified for each protein spot on the 2-DE polyacrylamide gel. The protein spots were classified into 11 groups according to protein function. Among the 11 groups, most protein spots of the DA(s) were identified as proteins related to metabolism. Additionally, unique protein spots of the DA(s) were related to abiotic stressors such as cold and heat. Those proteins are useful for improving wheat quality with resistance against abiotic stressors.

Texture Characteristics of Soybean-Curds Prepared with Different Coagulants and Compositions of Soybean-Curd Whey (응고제를 달리하여 제조한 두부의 텍스쳐 특성과 두부순물의 성분)

  • 이선미;황인경
    • Korean journal of food and cookery science
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    • v.13 no.1
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    • pp.78-85
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    • 1997
  • To determine the optimum coagulants concentrations for preparing soybean-curds, the transmittance of soybean-curd whey using spectrophotometer has been measured. The textural properties of soybean-curds were examined by texture analyzer and sensory evaluations. The general components, oligosaccharides and amino acids in soybean-curd wheys were analyzed. Protein patterns of soybean-curd wheys comparing with soyflour and soymilk were investigated. By texture analyzer, hardness, cohesiveness, springiness, and gumminess of Cacl$_2$ soybean-curd, MgCl$_2$ soybean-curd were higher than those of CaSO$_4$ soybean-curd and GDL soybean-curd. In the sensory evaluations, CaSO$_4$ soybean-curds and GDL soybean-curds were smoother and moister than others. Glutamic acid and aspartic acid were the first two abundant amino acids in three kinds of soybean-curd wheys, but arginine was the most abundant amino acid in GDL soybean-curd whey. Total sugar content of soybean-curd wheys were about 12-13 g/l, and the main sugars among 5 kinds of sugars were sucrose and raffinose. Electrophoresis using SDS-PAGE showed that glycinin and P-conglycinin, the main proteins of soybean appeared in soy flour and soymilk, and only low molecular weight subunits appeared in soybean-curd wheys.

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Production of Xylooligo-Saccharides and Purification of Extracellular Xylanase from Streptomyces chibaensis J-59 (방선균 Streptomyces chibaensis J-59 Xylanase의 정제 및 자일로 올리고당(Xylooligo-Saccharides)의 생산)

  • Joo, Gil-Jae;Rhee, In-Koo
    • Current Research on Agriculture and Life Sciences
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    • v.14
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    • pp.111-122
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    • 1996
  • S. chibaensis J-59 produced an extracellular xylanase in a CSL medium composed of 1.5% com steep liquor, 0.1% $MgSO_4{\cdot}7H_2O$, 0.012% $CoCl_2{\cdot}6H_2O$, and 0.15% glucose containing xylan. but it did not produce in the culture medium containing xylose. The production of enzyme reached to a maximum level (0.83 uints/ml) when bacteria were cultured in 2.5 l jar fermentor for 48hrs at $30^{\circ}C$ and pH 7.0. Furthermore, S. chibaensis J-59 produced an intracellular glucose isomerase in a medium containing xylan and/or xylose. Xylanase was purified 29-fold over the culture supernatants of S. chibaensis J-59 by ammonium sulfate fractionation, chromatography on DEAE-Sephadex A-50, and gel filtration on Sephadex G-200. The purified enzyme is a monomeric enzyme with a native molecular mass of 25 kDa and a subunit molecular mass of 25 kDa. The purified enzyme requires $Mg^{2+}$ for activity, $Ca^{2+}$, $Co^{2+}$ is not an inhibitor but inhibit by $Fe^{3+}$, $Hg^{2+}$, and $Cu^{2+}$, sodium dodecyl sulfate, N-bromosuccinide. Pattern of hydrolysis demonstrated that the xylanase was an endo-splitting enzyme able to break down birchwood xylan at random giving xylobiose, xylotriose and xylotetrose as the main end products.

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Gel and Texture Properties of Fish-meat Gel Prepared with Pagrus major in Comparison to Different Grades of Alaska Pollock (도미를 활용하여 제조한 연제품의 겔 및 texture 특성)

  • Gao, Ya;Oh, Jung Hwan;Karadeniz, Fatih;Lee, Seul-Gi;Kim, Hyung Kwang;Kim, Se Jong;Jung, Jun Mo;Cheon, Ji Hyeon;Kong, Chang-Suk
    • Journal of Life Science
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    • v.26 no.8
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    • pp.955-962
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    • 2016
  • Fish-meat gel is an intermediate product used in a variety of surimi-based seafood. One of the most-used raw materials of fish-meat gel is Alaska Pollock due to its high-quality meat in terms of gel strength and texture. However, increasing demand for fish-meat gel, along with overexploitation of the wild catch Alaska Pollock, has put the industry in need of low-cost sustainable alternative sources for fish-meat gel. Pagrus major (PM) is a widely aquacultured fish known for having white meat that is low in fat. The current study compares the quality of fish-meat gel prepared from aquacultured PM to that of high and mid-grade Alaska Pollock fish-meat gel. Gels were compared in terms of gel strength, texture, color, and protein pattern. Results indicated that fish-meat gels prepared from PM were superior to Alaska Pollock fish-meat gels with regard to gel strength, hardness, springiness, chewiness, cutting strength, and breaking force. In addition, although not matching in quality, PM exhibited a cohesiveness, whiteness, and expressible moisture content comparable to Alaska Pollock of both grades. Protein pattern analysis also showed that PM and Alaska Pollock fish-meat gels had similar protein profiles before and after gel preparation. Therefore, P. major is suggested as a potential substitute for Alaska Pollock in fish-meat gel production.

Physiological Changes of Saccharomyces cerevisiae KNU5377 Occurred in the Process of the 48-hour Ethanol Fermentation at 40℃ (40℃ 48시간 에탄올발효 과정 중 일어나는 Saccharomyces cerevisiae KNU5377의 생리 변화)

  • Kwak, Sun-Hye;Kim, Il-Sup;Kang, Kyung-Hee;Lee, Jung-Sook;Jin, Ingn-Yol
    • Journal of Life Science
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    • v.21 no.1
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    • pp.146-154
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    • 2011
  • In this study, physiological changes in a thermotolerant yeast Saccharomyces cerevisiae KNU5377 cell exposed to 48-hour alcohol fermentation at $40^{\circ}C$ were investigated. After 12 hours of alcohol fermentation at $40^{\circ}C$, the $C_{16:1}$ unsaturated acid of plasma membrane increased to 1.5 times more than the $C_{16:0}$ saturated fatty acid, and to about 2 times more for the $C_{18:1}$ unsaturated fatty acid. Fermentation at both $30^{\circ}C$ and $37^{\circ}C$ fermentation showed the same pattern as that done at $40^{\circ}C$. The pH of the alcohol-fermentation medium was reduced to pH 4.1 from a starting pH of 6.0 through the 12-hr fermentation and then maintained this level during the continuing fermentation. With the process of fermentation, the remaining glucose was reduced, but its amount remaining during the $40^{\circ}C$-fermentation was less reduced than those fermented at $30^{\circ}C$ and $37^{\circ}C$. In the study investigating the changing pattern of cellular proteins in the alcohol-fermenting cells, the SDS-PAGE and 2-D data indicated the most expressed dot was phosphoglycerate kinase, which is one enzyme involved in glycolysis. Why this enzyme was most expressed in the cells exposed to unfavorable conditions such as high temperature, increasing concentration of produced alcohol and long time exposure to other stress factors remains unsolved.

Plant Regeneration and Protein Analysis from Cadmium Resistant Callus of Tobacco (Nicotiana tabacum cv. BY4) (담배 (Nicotiana tabacum cv. BY4)카드뮴 저항성 캘러스로부터 식물체 재생과 단백질 분석)

  • 오승철;소웅영;조덕이;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.7-13
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    • 2001
  • Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.

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ELISA of paragonimiasis in cat by crude and purified antigens of Paragonimus westermani (폐흡충(肺吸蟲)의 조(粗)항원과 정제(精製)항원에 의한 폐흡충(肺吸蟲)감염 고양이혈청의 면역효소반응(免疫酵素反應))

  • Lee, Ok-Ran;Chang, Jae-Kyung
    • Parasites, Hosts and Diseases
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    • v.24 no.2
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    • pp.187-193
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    • 1986
  • Enzyme-linked immunosorbent assay(ELISA) using crude and affinity-purified antigens of adult worms of Paragonimus westermani was performed for infected cat sera with different worm burden, from preinfection to 18th week after infection. Crude antigen was used with supernatant of homogenated worms by freezing-thawing method, and the supernate was centrifuged for 1 hour at 10,000 rpm at $4^{\circ}C$. Affinity-purified antigen(antibody-bound antigen) was prepared from fractions(bound and unbound) of crude antigen by affinity chromatography on CNBr-activated sepharose 4B, and IgG as a ligand was prepared from paragonimiasis cat serum(6 months infected) obtained by ammonium sulfate ($40%{\sim}45%$ saturated) precipitation method. By SDS-PAGE, crude antigen showed 22 polypeptide fractions while purified antigen showed 4 fractions: 36, 400, 34, 700, 27, 600 and 11, 500 in molecular weights. All cats were divided into five groups($G_1-G_5$) by different worm burdens. The mean of recovered worms(${\pm}SD$) and the number of cats in each group are as follows: $G_1$, 2 worms(0) and 4 cats; $G_2$, 4.75 (${\pm}0.66$) and eight; $G_3$, 10.75(${\pm}1.92$) and four; $G_4$, 23.20(${\pm}3.43$) and five; $G_5$, 48(${\pm}12.63$) and five cats. The results were summarized as follows: 1. The antibody levels(OD value) increased by worm burden in $G_1$ to $G_4$ generally. However, individual antibody levels were not exactly related with worm burden in all groups, especially there was a wide difference in $G_4$ and $G_5$. These results suggested that the worm burden in $G_4$ (about $20{\sim}30$ worms) is enough to produce antibody maximum in cats of $2{\sim}3kg$ weight. 2. The antibody levels increased significantly(p<0.05) compared to control sera at the 3rd week in $G_1$ and $G_2$, at the 2nd week in $G_3$, and at the 1st week in $G_4$ and $G_5$. Especially in the 4th week, OD value increased more in $G_1$(p<0.01) and in $G_2$ to $G_5$(p<0.001). In the pattern of antibody levels by ELISA in each group, OD in $G_1$ increased to the 18th week continuously, in $G_2$ OD was maintained same after the 16th week, but in $G_3$ it decreased after the 16th week, and it was maintained same in $G_4$ and $G_5$ after the 14th week. 3. The antibody levels by ELISA with the affinity-purified antigen were higher than those with crude antigen in all groups generally. Especially, the difference of OD values between two antigens was larger from the 4th to the 10th week. In $G_1$ and $G_2$ OD with purified antigen was higher than that with crude one to the 18th week. It was also higher in $G_3$ than that with crude antigen to the 16th week and OD of $G_4$ and $G_5$ were higher before the 14th week than that with crude antigen, however became lower at the 16th week. Consequently, the antibody level in ELISA with affinity-purified antigen was more sensitive at the early weeks after infection and in light infection groups than that with crude antigen.

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