• Title/Summary/Keyword: sodium dodecyl sulfate (SDS)

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Expression of Pseudorabies Virus (PRV) Glycoproteins gB, gC and gD using Bacterial Expression System

  • Yun, Bit-Na-Rae;Bae, Sung-Min;Lee, Jun-Beom;Kim, Hee-Jung;Woo, Soo-Dong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.23 no.1
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    • pp.147-153
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    • 2011
  • The Pseudorabies (PR), also called Aujeszky's disease (AD), is an infectious viral disease caused by an alpha herpes virus and has domestic and wild pigs, as well as a wide range of domestic and wild animals, as the natural host. Pseudorabies virus (PRV) virions contain several envelope glycoproteins. Among them, gB, gC and gD are regarded as the major immunogenic proteins. We expressed these glycoproteins using the bacterial expression system and analyzed recombinant proteins. Expression of glycoproteins gC and gD were observed on SDS-PAGE or Western blot analysis, but gB was not. Optimal concentration of IPTG and inducing time were determined as 1.0 mM and 4 h, respectively, for the expression of both gC and gD in E. coli. A sodium dodecyl sulfate (SDS) was the most efficient detergent in solubilizing insoluble recombinant protein.

An Extraction of Barley Protein and a Comparison of the Protein Composition of Some Barleys -Electrophoretic Pattern of Barley Protein- (보리단백질의 추출 및 품종간 조성비교 -II. 보리단백질의 품종간 조성비교-)

  • Kim, Ze-Uook;Kim, Jung-Sang
    • Applied Biological Chemistry
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    • v.29 no.1
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    • pp.57-61
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    • 1986
  • Total protein from 4 barley varieties(Olbori, Young San-bori, Sacheon 6, and Suwon 228) was separated into albumin, globulin hordein and glutelin fractions by the Osborne method and tile modified Osborne method in a previous report. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for the protein fractions revealed that there was a little difference in the polypeptide composition of each fraction among four varieties. The comparision of hordeins and hordein-Is by polyacrylamide gel electrophoresis at pH 3.0 showed a marked difference among the varieties. Hordein-I contained a high level of glutamic acid and proline and low level of lysine. And (here was little difference in amino acid composition of hordein-Is which were extracted from the 4 varieties.

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Characterization of Edible Film Fabricated with Channel Catfish Ictalurus punctatus Gelatin by Cross-Linking with Transglutaminase

  • Oh, Jun-Hyun
    • Fisheries and Aquatic Sciences
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    • v.15 no.1
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    • pp.9-14
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    • 2012
  • The objectives of this research were to improve the film-forming properties of Channel Catfish Ictalurus punctatus skin gelatin (CSG) by cross-linking with transglutaminase (TG), determine and optimize the TG reaction time, and characterize the mechanical and barrier properties of CSG edible film. Cross-linking of CSG was performed by TG for 0, 10, 20, 30, and 40 min at $50^{\circ}C$, and the reaction was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The color and mechanical and barrier properties of edible films fabricated with CSG cross-linked with TG were characterized. Gelatin yields from the extraction ranged from 18.2% to 23.3%. SDS-PAGE exhibited dark bands at 120 and 250 kDa, indicating successful TG-mediated cross-linking. The color of CSG film was not affected by TG cross-linking. The tensile strength of CSG films cross-linked with TG decreased from 42.59 to 21.73 MPa and the percent elongation increased from 42.92% to 76.96% as reaction times increased from 0 to 40 min. There was no significant difference in water vapor permeability of CSG films.

Effect of Ginseng Saponins on $K^+-Dependent$ Phosphatase Activity of Dog Cardiac Sarcolemma (인삼 사포닌이 개 심실 형질막의 $K^+$-의존성 포스파타제 활성에 미치는 영향)

  • Lee, Shin-Woong;Lee, Jeung-Soo
    • YAKHAK HOEJI
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    • v.36 no.2
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    • pp.129-136
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    • 1992
  • The effects of ginseng saponins, gypsophila saponin, sodium dodecyl sulfate(SDS), and Triton X-100 on membrane $K^+-dependent$ phosphatase activity which is lipid dependent and represents dephosphorylation step of the complete Na+, $K^+-ATPase$ reaction were investigated in this study to elucidate whether the effects of ginseng saponins are due to the detergent action, using sarcolemma enriched preparation isolated from dog ventricle. $Na^+$, $K^+-ATPase$ and $K^+-dependent$ phosphatase activities of cardiac sarcolemma were about $143\;{\mu}mol$ Pi/mg protein/hr and $34\;{\mu}mol$ p-nitrophenol/mg protein/hr, respectively. While ginseng saponins (triol>total>diol) inhibited $K^+-dependent$ phosphatase activity, gypsophila saponin, and low dose of SDS($0.4\;{\mu}g/{\mu}g$ protein), and Triton X-100 ($0.6\;{\mu}g/{\mu}g$ protein) increased the enzyme activity, indicating disruptive effect of detergents on membrane barriers. The activating effect of low doses of Triton X-100 on membrane $K^+-dependent$ phosphatase appeared at concentration decreasing light scattering. However, the inhibitory effect of ginseng saponin appeared before a decrease in light scattering. These results suggest that low concentrations of ginseng saponins inhibit the membrane $K^+-dependent$ phosphatase by interacting directly with enzyme before membrane disruption.

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The Synthesis and Optical Properties of Silica Coated CdSe/ZnS QDs (실리카가 코팅된 양자점의 코팅두께에 따른 광 특성 변화)

  • Lee, Ji-Hye;Shin, Hyun Ho;Lee, Jong-Heun;Hyun, Sang Il;Koo, Eunhae
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.26 no.3
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    • pp.221-226
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    • 2013
  • The water soluble quantum dots (QDs) are synthesized by the phase transfer and silica coating reaction. The photoluminescence intensity of silica-coated QDs are mainly affected by the amount of phase transfer agent, SDS (sodium dodecyl sulfate), and the maximum value is obtained at the cmc (critical micell concentration) concentration of SDS in the phase transfer reaction. Based on fluorescence spectra and field emission transmission electron microscope (FETEM), the energy transfer rate by forster resonance energy transfer (FRET) is increasing with the thickness of the silica shell coated on CdSe/ZnS QDs.

Quality properties of whole milk powder on chicken breast emulsion-type sausage

  • Kang, Kyu-Min;Lee, Sol-Hee;Kim, Hack-Youn
    • Journal of Animal Science and Technology
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    • v.63 no.2
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    • pp.405-416
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    • 2021
  • The aim of the study was to determine the effect of whole milk powder (WMP) as heterologous proteins on chicken breast emulsion-type sausages. The quality properties of WMP on such chicken breast emulsion-type sausages were investigated by measuring the proximate composition, pH, color, cooking yield, protein solubility, and by applying other methods, such as texture profile analysis (TPA), microphotograph, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and electronic nose. The crude fat, protein, and ash contents of 15% WMP samples were significantly higher than the control samples (p < 0.05). The redness of the cooked samples significantly increased with an increase in the WMP contents (p < 0.05). The cooking yield of WMP treated samples was significantly higher than the control sample (p < 0.05). Additionally, the hardness, gumminess, and chewiness of WMP treated samples were significantly higher than the control sample (p < 0.05). The sarcoplasmic and myofibrillar proteins of samples containing 15% WMP were significantly higher than the control samples (p < 0.05). The result of SDS-PAGE showed that the C protein, sarcoplasmic protein, actin, and tropomyosin increased with an increase in the WMP contents. The principal component analysis plot of WMP-treated samples was clearly different from that of the control samples. Based on these results, it was predicted that WMP could be useful as heterologous protein on emulsion-type sausage.

Purification, Characterization and Application of a Cold Active Lipase from Marine Bacillus cereus HSS

  • Hassan, Sahar WM.;Abd El Latif, Hala H.;Beltagy, Ehab A.
    • Microbiology and Biotechnology Letters
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    • v.50 no.1
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    • pp.71-80
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    • 2022
  • Lipases (triacylglycerol acylhydrolases [EC 3.1.1.3]) are water-soluble enzymes. They catalyze the hydrolysis of fats and oils. A cold-active lipase from marine Bacillus cereus HSS, isolated from the Mediterranean Sea, Alexandria, Egypt, was purified and characterized. The total purification depending on lipase activity was 438.9 fold purification recording 632 U/mg protein. The molecular weight of the purified lipase was estimated to be 65 kDa using sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The optimum substrate concentration, enzyme concentration, pH, and temperature were 1.5 mM, 100 µl, pH 6 and 10℃, respectively. The lipase was tolerant to NaCl concentrations ranging from 1.5 to 4.5%. The lipase was affected by the tested metal ions, and its activity was inhibited by 16% in the presence of 0.05 M SDS. The application of the cold-active lipase for the removal of an oil stain from a white cotton cloth showed that it is a promising biological agent for the treatment of oily wastes and other related applications. To the best of our knowledge, this is the first report of the purification and characterization of a lipase from marine B. cereus HSS isolated from the Mediterranean Sea.

Effects of Using Soybean Protein Emulsion as a Meat Substitute for Chicken Breast on Physicochemical Properties of Vienna Sausage

  • Kang, Kyu-Min;Lee, Sol-Hee;Kim, Hack-Youn
    • Food Science of Animal Resources
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    • v.42 no.1
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    • pp.73-83
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    • 2022
  • The aim of this study is to determine the effects of using emulsion manufactured with soybeans (ES) to substitute chicken breast in Vienna sausages. Four types of Vienna sausages (S1: 10% ES and 50% chicken, S2: 20% ES and 40% chicken, S3: 30% ES and 30% chicken, and S4: 40% ES and 20% chicken) for this study were made. The pH, color, proximate composition, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), microphotographs, cooking yields, and texture profile analysis of sausages were examined. The pH value of uncooked and cooked sausages increased significantly with increasing ES content (p<0.05). The crude protein contents of S2, S3, and S4 were significantly higher than that of the control (p<0.05). Furthermore, the SDS-PAGE results showed that α-conglycinin, β-conglycinin, and the acidic subunit of glycinin all increased with increasing ES content. Microphotographs revealed that increasing the ES content decreased the size of fat globules. The cooking yields of samples increased significantly with increasing ES content (p<0.05). The hardness values of ES treated samples were significantly lower than that of the control (p<0.05). Therefore, 30% substitute of chicken breast with ES can improve the quality and structure of Vienna sausage, without inducing critical defects.

Purification and Characterization of a Fibrinolytic Enzyme from Bacillus pumilus 2.g Isolated from Gembus, an Indonesian Fermented Food

  • Afifah, Diana Nur;Sulchan, Muhammad;Syah, Dahrul;Yanti, Yanti;Suhartono, Maggy Thenawidjaja;Kim, Jeong Hwan
    • Preventive Nutrition and Food Science
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    • v.19 no.3
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    • pp.213-219
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    • 2014
  • Bacillus pumilus 2.g isolated from gembus, an Indonesian fermented soybean cake, secretes several proteases that have strong fibrinolytic activities. A fibrinolytic enzyme with an apparent molecular weight of 20 kDa was purified from the culture supernatant of B. pumilus 2.g by sequential application of ammonium sulfate precipitation, ion-exchange chromatography, and hydrophobic chromatography. The partially purified enzyme was stable between pH 5 and pH 9 and temperature of less than $60^{\circ}C$. Fibrinolytic activity was increased by 5 mM $MgCl_2$ and 5 mM $CaCl_2$ but inhibited by 1 mM phenylmethylsulfonyl fluoride (PMSF), 1 mM sodium dodecyl sulfate (SDS), and 1 mM ethylenediaminetetraacetic acid (EDTA). The partially purified enzyme quickly degraded the ${\alpha}$ and ${\beta}$ chains of fibrinogen but was unable to degrade the ${\gamma}$ chain.

Purification and Characterization of a Keratinase from a Feather-Degrading Fungus, Aspergillus flavus Strain K-03

  • Kim, Jeong-Dong
    • Mycobiology
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    • v.35 no.4
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    • pp.219-225
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    • 2007
  • A keratinolytic enzyme secreted by Aspergillus flavus K-03 cultured in feather meal basal medium (FMBM) containing 2% (w/v) chicken feather was purified and characterized. Keratinolytic enzyme secretion was the maximal at day 16 of the incubation period at pH 8 and $28^{\circ}C$. No relationship was detected between enzyme yield and increase of fungal biomass. The fraction obtained at 80% ammonium sulfate saturation showed 2.39-fold purification and was further purified by gel filtration in Sephadex G-100 followed by ion exchange chromatography on DEAE-Sephadex A-50, yielding an active protein peak showing 11.53-fold purification. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and zymograms indicated that the purified keratinase is a monomeric enzyme with 31 kDa molecular weight. The extracellular keratinase of A. flavus was active in a board range of pH ($7{\sim}10$) and temperature ($30^{\circ}C{\sim}70^{\circ}C$) profiles with the optimal for keratinase activity at pH 8 and $45^{\circ}C$. The keratinase activity was totally inhibited by protease inhibitors such as phenylmethylsulfonyl fluoride (PMSF), iodoacetic acid, and ethylenediaminetetraacetate (EDTA) while no reduction of activity by the addition of dithiothreitol (DTT) was observed. N-terminal amino acid sequences were up to 80% homologous with the fungal subtilisins produced by Fusarium culmorum. Therefore, on the basis of these characteristics, the keratinase of A. flavus K-03 is determined to be subtilisins-like.