• Title/Summary/Keyword: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Chemical composition of protein concentrate prepared from Yellowfin tuna Thunnus albacares roe by cook-dried process

  • Lee, Hyun Ji;Park, Sung Hwan;Yoon, In Seong;Lee, Gyoon-Woo;Kim, Yong Jung;Kim, Jin-Soo;Heu, Min Soo
    • Fisheries and Aquatic Sciences
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    • v.19 no.3
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    • pp.12.1-12.8
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    • 2016
  • Roe is the term used to describe fish eggs (oocytes) gathered in skeins and is one of the most valuable food products from fishery sources. Thus, means of processing are required to convert the underutilized yellowfin tuna roes (YTR) into more marketable and acceptable forms as protein concentrate. Roe protein concentrates (RPCs) were prepared by cooking condition (boil-dried concentrate, BDC and steam-dried concentrate, SDC, respectively) and un-cooking condition (freeze-dried concentrate, FDC) from yellowfin tuna roe. The yield of RPCs was in the range from 22.2 to 25.3 g/100 g of roe. RPCs contained protein (72.3-77.3 %), moisture (4.3-5.6 %), lipid (10.6-11.3 %) and ash (4.3-5.7 %) as the major constituents. The prominent amino acids of RPCs were aspartic acid, 8.7-9.2, glutamic acid, 13.1-13.2, and leucine, 8.5-8.6 g/100 g of protein. Major differences were not observed in each of the amino acid. K, S, Na, and P as minerals were the major elements in RPCs. No difference noted in sodium dodecyl sulfate polyacrylamide gel electrophoresis protein band (15-100 K) possibly representing partial hydrolysis of myosin. Therefore, RPCs from YTR could be use potential protein ingredient for human food and animal feeds.

Effect of Recombinant Olive Flounder Stanniocalcin on Serum Calcium Levels (혈청 칼슘 농도에 미치는 넙치 유전자 재조합 스타니오칼신의 효과)

  • Shin, Ji-Hye;Jung, Yu-Jung;Han, Yoon-Hee;Lee, Kyun-Young;Lee, Kyung-Mi;Kaneko, Toyoji;Sohn, Young-Chang
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.4
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    • pp.307-313
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    • 2010
  • Stanniocalcin 1 (STC1) is a glycoprotein hormone that is important in the maintenance of calcium and phosphate homeostasis in both fish and mammals. STC1 and its paralog STC2 are expressed in multiple tissues in fishes, although the physiological roles of piscine STCs are still unclear compared with those of mammals. In this study, we cloned olive flounder STC1 (ofSTC1) and ofSTC2 cDNAs into pET28a vector and used E. coli Rosetta (DE3) as the host strain for protein expression. Expression experiments were carried out using isopropyl-$\beta$-D-thiogalactoside (IPTG) and nickel affinity chromatography. We could identify the recombinant proteins as single 29.5 kDa (ofSTC1) and 33.2 kDa (ofSTC2) bands in the insoluble fraction on sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE). These results indicate that ofSTC1 and ofSTC2 were expressed as insoluble proteins in E. coli. Furthermore, the injection of ofSTC1 protein into juvenile tilapia resulted in a decrease of the serum calcium level. These results suggest that the purified fish STC1 and STC2 proteins may be used to elucidate the physiological role of STCs in fishes.

Comparative effects of dry-aging and wet-aging on physicochemical properties and digestibility of Hanwoo beef

  • Kim, Ji-Han;Kim, Tae-Kyung;Shin, Dong-Min;Kim, Hyun-Wook;Kim, Young-Boong;Choi, Yun-Sang
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.3
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    • pp.501-505
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    • 2020
  • Objective: The purpose of this study was to investigate the effects of aging methods (AM) i.e. dry-aging (DA) and wet-aging (WA) on the physicochemical properties and in vitro digestibility of proteins in beef short loin. Methods: Short loins (M. longissmus lumborum), were trimmed and boned-out on the fifth day postmortem, from a total of 18 Hanwoo, which were purchased from a commercial slaughterhouse. Short loins were separated randomly grouped into one of the three treatments: control, WA (1℃, 7 days), and DA (1℃, 0.5 m/s, 85% relative humidity [RH], 30 days). Results: Dry-aged beef (DAB) exhibited higher pH, water holding capacity (WHC), myofibrillar fragmentation index (MFI), and digestibility, however lower lightness, redness, and yellowness values, cooking loss, and shear force (SF), than those of wet-aged beef (WAB) (p<0.05). The myosin light chain band intensity of DAB was higher than that of control and WAB in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The in vitro digestibility of aged beef was highly (p<0.001) correlated to physicochemical properties except WHC. The correlation coefficient between AMs and WHC was higher than that between AM and SF (p<0.05) or MFI (p<0.001). A high correlation was observed between SF and MFI (p<0.001). Conclusion: Thus, we believe that DAB is more advantageous than WAB owing to its high digestibility and WHC and low SF.

Purification and Properties of Extracellular Cytosine Deaminase from Chromobacterium violaceum YK 391

  • Yu, Tae-Shick;Kim, Tae-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.173-178
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    • 1999
  • The extracellular cytosine deaminase (EC 3.5.4.1) from Chromobacterium violaceum YK 391 was purified 264.7-fold with an overall yield of 14.3%. The enzyme was for the first time homogeneous by the criteria of polyacrylamide gel electrophoresis performed in the absence and in the presence of sodium dodecyl sulfate. The molecular weight of the purified enzyme was estimated to be about 156 kDa. The enzyme consisted of two identical subunits of approximate molecular weight 78 kDa. The isoelectric point of the enzyme was pH 5.55. The enzyme had a pH optimum of 7.5 and a temperature optimum of around 40 to $45^{\circ}C$. Besides cytosine, the enzyme deaminated 5-fluorocytosine, cytidine, 5-methylcytosine, and 6-azacytosine, but not 5-azacytosine. The extracellular cytosine deaminase is believed to be unique because it was active not only on cytosine but also on cytidine. The apparent $K_m$ values for cytosine, 5-fluorocytosine, cytidine, and 5-methylcytosine were determined to be 1.55 mM, 5.52 mM, 10.4 mM, and 67.2 mM, respectively. The enzyme activity was strongly inhibited by heavy metal ions such as $Fe^{2+},Pb^{2+},Cd^{2+},Zn^{2+}, Hg^{2+}, and Cu^{2+}$ at 1 mM, and completely by $\alpha,\alpha$'-dipyridyl, and $\rho$-chloromercuribenzoate at 1 mM, and weakly inhibited by 1mM ο-phenanthroline. The enzyme activity was not affected by various nucleosides and nucleotides.

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Development of an Enzyme-linked Immunosorbent Assay Using Vitellin for Vitellogenin Measurement in the Pale Chub, Zacco platypus

  • Lim, Eun-Suk;Lee, Eun Hee;Kim, Myung Hee;Han, Chang-Hee;Lee, Sung-Kyu;Kim, Jiwon
    • Environmental Analysis Health and Toxicology
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    • v.28
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    • pp.16.1-16.8
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    • 2013
  • Objectives Fish vitellogenin (VTG) is produced in the female liver during oogenesis through the estradiol cycle and produced in the male liver by endocrine disrupting chemicals (EDCs) such as alkylphenols. In this study, we propose that the VTG concentration in the pale chub could be detected using monoclonal antibodies and polyclonal antibodies against vitellin (Vn) in a VTG enzyme-linked immunosorbent assay (ELISA) system. Methods Monoclonal antibodies and polyclonal antibodies were produced using the Vn extracted from the matured ovum of the ovary. The VTG was extracted from the plasma of the male pale chub. The Vn and VTG were confirmed by measuring the molecular weight of their proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the specificity of the antibodies was checked through western blotting methods. The assay system was validated with respect to optimal assay concentrations, specificity, recovery, and intra- and inter-assay variations. Results The Vn consisted of two protein bands with apparent molecular weights of 64 and 37 kDa. The SDS-PAGE indicated protein weights of 146 and 77 kDa in the VTG. The assay range was 15.6 ng/mL to 2,000 ng/mL, and the value of the intra- and inter-assay variations were within 10.0% and 14.7%, respectively. The recovery rate was $99.5{\pm}5.5%$. Conclusions A sandwich ELISA was developed that could be used to qualify the VTG of pale chub in screening for EDCs. Pale chub is an ideal species for observing estrogen activity in the environment because of its extensive habitat and extensive food chain. The ELISA developed here would be more favorable than those for other species for determining the effect of long-term food chain accumulation of EDCs in aquatic environments.

Collection, Conservation, and Utilization of Plant Genetic Resources -emphasis on weeds- (유용식물 유전자원의 수집 보존 및 이용에 관한 연구 -잡초를 대상으로-)

  • Kim, Kil Ung
    • Current Research on Agriculture and Life Sciences
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    • v.8
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    • pp.9-18
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    • 1990
  • Germinating ability of 54 weed species collected throughout Korea in 1987 and 1988 was determined in incubator at $25^{\circ}C$ for 15 days. The seeds of Echinochloa sp, collected from 70 different places throughout Korea, were classified by Yabuno's seed morphology method. Plant characters of two identified Echinochloa species, one having two subspecies were evaluated. SDS-PAGE(sodium dodecyl sulfate polyacrylamide gel electrophoresis) of seed protein assay was used to identify locally collected Echinochloa species classified by Yabuno's seed morphology method. 1. Out of 54 weed species collected, 31 species showed germination at 15 days after incubation, and 14 species showed over 50% and no germination was observed in 24 species, indicating that approximately 40 species had varied degree of germinating ability. 2. It was confirmed that two annual species of Echinochloa such as E. oryzicola and E. crus-galli occurred in Korea and one of which particularly E. crus-galli has two subspecies such as E. c. var. praticola and E. c. var. crus-galli. 3. E. oryzicola was distinguished from E. c. var crus-galli and E. c. var praticola by presence or absence of one major protein band in B zone. However, it was rather difficult to distinguish E. crus-galli species because they showed similar protein pattern.

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Effects of Psychrotrophic Bacteria, Serratia liquefaciens and Acinetobacter genomospecies 10 on Yogurt Quality

  • Shin, Yong Kook;Oh, Nam Su;Lee, Hyun Ah;Choi, Jong-Woo;Nam, Myoung Soo
    • Food Science of Animal Resources
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    • v.34 no.4
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    • pp.543-551
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    • 2014
  • The aim of this study was to evaluate the effect of proteolytic (Serratia liquefaciens, match %: 99.39) or lipolytic (Acinetobacter genomospecies 10, match %: 99.90) psychrotrophic bacteria (bacterial counts, analysis of free fatty acids (FFA) and analysis of free amino acids) on the microbial and chemical properties (yogurt composition), and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of yogurt during storage. Yogurts were prepared with raw milk preinoculated with each psychrotrophic bacteria. The total solid, fat, and protein content were not affected by preinoculation, but the pH of yogurt preinoculated with psychrotrophic bacteria was higher than in control. There was a dramatic increase in short chain free fatty acids among FFA in yogurt with Acinetobacter genomospecies 10. For 14 d of cold storage condition, SCFFA was 25.3 mg/kg to 34.4 mg/kg (1.36 times increased), MCFFA was 20.4 mg/kg to 25.7 mg/kg (1.26 times increased), and LCFFA was 240.2 mg/kg to 322.8 mg/kg (1.34 times increased). Serratia liquefaciens (match %: 99.39) in yogurt caused a greater accumulation of free amino acids (FAA), especially bitter peptides such as leucine, valine, arginine, and tyrosine, but SDS-PAGE showed that the inoculation of Serratia liquefaciens did not affect the degree of casein degradation during storage. Taken together, the excessive peptides and FFA in yogurt generated from psychrotrophic bacteria could develop off-flavors that degrade the quality of commercial yogurt products.

Marination and Physicochemical Characteristics of Vacuum-aged Duck Breast Meat

  • Khan, Muhammad Issa;Lee, Hyun Jung;Kim, Hyun-Joo;Young, Hae In;Lee, Haelim;Jo, Cheorun
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.11
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    • pp.1639-1645
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    • 2016
  • We investigated marinade absorption and physicochemical characteristics of vacuum-aged duck breasts that were halved and individually vacuum-packed for chiller aging at $4^{\circ}C$ for 14 d. One half was marinated for 0, 7, or 14 d, while the second half was used as a control. Marinade absorption, cooking loss, cooking yield, texture profile, pH, color, protein solubility, and thiobarbituric acid reactive substances (TBARS) values were evaluated, and protein sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed. Marinade absorption and pH did not vary significantly after 14 d of aging. Marination increased the pH, color ($a^*$ and $b^*$) values, and cooking yield and reduced cooking loss. TBARS values significantly increased with aging time, but were significantly reduced by marination. Myofibril and total protein solubility increased with aging and marination, while SDS-PAGE showed protein degradation. Hence, aging and marination can be used simultaneously to improve physicochemical quality and cooking yield of vacuum-aged duck breast.

A Promising Serum Autoantibody Marker, Anti-Heat Shock Protein 90α, for Cholangiocarcinoma

  • Boonjaraspinyo, Sirintip;Juasook, Amornrat;Boonmars, Thidarut;Aukkanimart, Ratchadawan;Silsirivanit, Atit;Loilome, Watcharin;Sriraj, Pranee;Wu, Zhiliang;Ratanasuwan, Panaratana
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.14
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    • pp.5779-5785
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    • 2015
  • The present study was designed to investigate cholangiocarcinoma (CCA) antibodies in hamster serum. Hamster CCA cell lines were processed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. A candidate biomarker was confirmed by immunoprecipitation and western blot, and was further analyzed using ELISA and sera from normal control hamsters, hamsters with opisthorchiasis and hamsters with various stages of CCA, as well as from CCA patients and healthy individuals. One candidate marker was identified as $HSP90{\alpha}$, as indicated by a high level of anti-$HSP90{\alpha}$ in hamster CCA sera. It was found that the levels of anti-$HSP90{\alpha}$ were specifically elevated in the sera of hamsters with CCA compared with other groups and progressively increased with the clinical stage. At the cut-off point of 0.4850 on the receiver operating characteristic curve, anti-$HSP90{\alpha}$ could discriminate CCA from healthy control groups with a sensitivity of 76.2%, specificity of 71.4% and total accuracy 75.5%. In the present study, we have shown that anti-$HSP90{\alpha}$ may be a potential useful serum biomarker to discriminate CCA cases from healthy persons.

Functionality Changes of Rapeseed Protein upon Proteolysis (유채단백질의 Proteolysis에 의한 기능성 변화)

  • Kim, Chung-Hee;Kim, Hyo-Sun;Lee, Jang-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.5
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    • pp.519-524
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    • 1992
  • purified rapeseed(Brassica napus var. Youngsan) protein was hydrolyzed by pronase. The hyrolysate protein was investigated for the some physicochemical and functional properties. UV and intrinsic fluorescence spectra of the hydrolysate showed the maximum absorption at 274nm and 360nm respectively. Intensity of yellow color decreased in the process of hydrolysis and the surface hydrophobicity decreased up to fourfold. The main bands of hydrolysate by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) were observed at 14,000 to 12,000 dalton molecular weight. Solubilities of hydrolyzed protein increased by 10~15% compared to those of unhydrolyzed protein at acidic pH. In the hydrolysate, while absorption of both water and oil, foam expansion and emulsion stability were increased, absolute viscosity, heat coagulation, calcium coagulation, foam stability and emulsion activity were decreased.

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