• Title/Summary/Keyword: Soluble protein

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Influence of Site-Directed Mutagenesis on Protein Assembly and Solubility of Tadpole H-chain Ferritin

  • Kim, Kyung-Suk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.2
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    • pp.67-70
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    • 1998
  • In order to understand the influence of ferroxidase center on the protein assembly and solubility of tadpole ferrin, three mutant plasmids, pTH58K, pTH61G, and pTHKG were constructed with the aid of site-directed mutagenesis and mutant proteins were produced in Eshcerichia coli. Mutant ferritin H-subunits produced by the cells carrying plasmids pTH58K and pTHKG were active soluble proteins, whereas the mutant obtained from the plasmid pTH61G was soluble only under osmotic stress in the presence obtained from the plasmid pTH61G was soluble only under osmotic stress in the presence of sorbitol and betaine. Especially, the cells carrying pTH61G together with the plasmid pGroESL harboring the molecular chaperone genes produced soluble ferritin. The mutant ferritin H-subunits were all assembled into ferritin-like holoproteins. These mutant ferritns were capable of forming stable iron cores, which means the mutants are able to accumulate iron with such modified ferroxidase sites. Further functional analysis was also made on the individual amino acid residues of ferroxidase center.

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Protein Profiles of Major Korean Rice Cultivars

  • Wei, Chuanha;Kwon, Oh-Yun;Liu, Xi-Wen;Kim, Hyoung-Chin;Yoon, Won-Kee;Kim, Hwan-Mook;Kim, Mee-Ree
    • Preventive Nutrition and Food Science
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    • v.12 no.2
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    • pp.103-110
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    • 2007
  • The protein profiles among Korean rice cultivars were assessed by total protein determination, solubility fractionation, SDS-PAGE analysis and scanning densitometry. In the extraction of protein, the SDS/urea system at a neutral pH was more efficient than that at alkaline pH. The determination of total protein showed that the protein content was similar among cultivars, ranging from 87.9 to 92.7 mg/g dry weight. Additionally, the water/NaCl-soluble protein fraction, containing 14${\sim}$16 kDa albumin and 22 kDa globulin ${\alpha}$-globulin, was also similar among cultivars, with a range of 9.94 to 11.98 mg/g dry weight. The SDS-PAGE/densitometry of total protein showed that there was no discernable difference in proteins of higher molecular weights among various cultivars, whereas the amount of lower molecular weight proteins (14${\sim}$16 kDa) is somewhat variable among cultivars. Furthermore, SDS-PAGE analysis of water/NaCl-soluble and propanol-soluble fractions indicates that there is a discernible change in the content of albumin, globulin or prolamin among cultivars. Thus, the PAGE/densitometry method, preceded by solubility fractionation, is useful for examining differences in protein profiles of rice cultivars.

Interactions between Chicken Salt-soluble Meat Proteins and Makgeolli Lees Fiber in Heat-induced Gels

  • Choi, Yun-Sang;Park, Kwoan-Sik;Kim, Hack-Youn;Kim, Hyun-Wook;Song, Dong-Heon;Chung, Hai-Jung;Lee, Ju-Woon;Kim, Cheon-Jei
    • Food Science of Animal Resources
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    • v.31 no.6
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    • pp.817-826
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    • 2011
  • The technological effects of Makgeolli lees fiber (0, 0.5, 1.0, 2.0, and 4.0%) on chicken salt-soluble breast meat proteins in a model system on proximate composition, physicochemical properties, and textural properties were investigated. Makgeolli lees fiber was obtained from Makgeolli brew processing, and the by-products showed good dietary fiber. The moisture and ash contents, water holding capacity, redness, yellowness, hardness, and apparent viscosity of chicken salt-soluble meat protein heat-induced gel systems with Makgeolli lees fiber were all higher than the control without Makgeolli lees fiber. However, protein solubility and electrophoretic patterns did not differ among the control and treatments with Makgeolli lees fiber samples. The chicken salt-soluble protein heat-induced gel systems incorporating Makgeolli lees fiber had improved water holding capacity, textural properties, and viscosity due to Makgeolli lees fiber addition. These results suggest that the addition of 4.0% Makgeolli lees fiber to gel is helpful to improve the physical properties of heat-induced gels.

Investigation into the Distribution of Total, Free, Peptide-bound, Protein-bound, Soluble-and Insoluble-Collagen Hydroxyproline in Various Bovine Tissues

  • Siddiqi, Nikhat J.;Alhomida, Abdullah S.
    • BMB Reports
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    • v.36 no.2
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    • pp.154-158
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    • 2003
  • Collagen is a family of proteins which consists of several genetically distinct molecular species and is intimately involved in tissue organization, function, differentiation and development. The purpose of this study was to investigate the concentration of different hydroxyproline (Hyp) fractions viz., total, free, peptide-bound, protein-bound, soluble- and insoluble-collagen hydroxyproline (Hyp) in various bovine tissues. Results showed that liver had the highest concentration of free Hyp followed by kidney, brain, spleen, lungs, muscle and heart. Liver also had the highest concentration of peptide-bound collagen Hyp followed by kidney, heart, spleen, lungs, brain and muscle. The concentration of protein-bound collagen Hyp was highest in the liver, followed by kidney, spleen, lungs, muscle, brain and heart. Total Hyp was highest in the liver, followed by kidney, spleen, brain, heart, muscle and lungs. Liver also had significantly high concentration of collagen as compared to other tissues examined (P<0.001). Spleen had the significantly higher concentration of soluble-collagen Hyp when compared to other tissues (P<0.001). This was followed by heart, muscle, lungs, brain, kidney and liver. Heart had the highest concentration of insoluble-collagen Hyp followed by lungs, kidney, liver, muscle, spleen and brain. The variation among the insoluble-collagen Hyp concentration of heart and muscle, spleen and brain was significant (P<0.001). We speculate that these differences could be due to the variation in turn over of rate of collagen metabolism in this species.

Processing, Quality Stability and Utilization of Approved Sardine Surimi for Surimi-based Products (정어리 개량고기풀의 제조와 품질 안정성 및 이용성)

  • 김태진;서상복;이두석;민진기
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.2
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    • pp.403-408
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    • 1999
  • This study was conducted to investigate chemical properties, quality stability and utilization of approved sardine surimi(ASS) which is developed our laboratory. The product yield of the ASS was about 3 times higher than that of usual sardine surimi(SS). The proper addition concentration of sodium bicarbonate was 0.1% for the neutralization of the ASS. The content of salt soluble protein nitrogen in the ASS was about the half of that in the SS, while the content of water soluble protein nitrogen was 2.4 times higher in ASS. The total amount of free amino acids in the ASS was about 11 times higher than that of the SS. Predominant free amino acids in the ASS were histidine, taurine, glutamic acid and alanine, and those four amino acids occupied 94% of total amount of free amino acids. During cold storage at 21oC for 6 months, the quality of ASS was more stable than that of SS in judging from changes of water soluble and salt soluble protein nitrogen, AV and POV. Quality of fish burger, fish sausage and fried fish paste processed in accordance with commercial processing preparation using the ASS or SS exclusively and mixtures which other white meat fish surimi(alaska pollack, hair tail and sole) were proportionallly added to each of two types of sardine meat were evaluated. In case of fish burger, the product processed from the ASS only were superior.

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Studies on the Composition of Fatty Acid and Protein in Pumpkin Seeds (호박씨의 지방산(脂肪酸) 및 단백질(蛋白質)의 조성(組成)에 관(關)한 연구(硏究))

  • Kim, Jun-Pyong;Lee, Young-Ja;Namkung, Sok
    • Korean Journal of Food Science and Technology
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    • v.10 no.1
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    • pp.83-87
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    • 1978
  • Dehulled pumpkin seeds produced in Korea were defatted and their composition of fatty acid was analyzed by gas chromatography. Soluble protein and insoluble protein were isolated from the dehulled and defatted pumpkin seeds. The protein component was identified by disc-electrophoresis and the amino acid composition of the protein was analyzed. (1) The pumpkin seed contained 46.5% of fat and 25.5% of protein respectively. (2) The fatty acid of the pumpkin seed was compassed chiefly of linoleic acid (51.74%), oleic acid (22.2%), palmitic acid (18.1%) and stearic acid (7.84%) (3) The amino acid composition of the soluble and insoluble seed protein contained most of all the amino acids except for S-contained amino acids. (4) It has been identified by disc-electrophoresis that the soluble protein had 13 bands and the insoluble protein had 4 bands.

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Modulation of the Tendency Towards Inclusion Body Formation of Recombinant Protein by the Addition of Glucose in the araBAD Promoter System of Escherichia coli

  • Lee, You-Jin;Jung, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1898-1903
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    • 2007
  • We attempted to modulate the overall protein expression rate through the addition of a repressor against the araBAD promoter system of Escherichia coli, in which glucose was used as a repressor. Therefore, 0.5% L-arabinose was initially contained as an inducer in culture medium, and either 2% glucose or 2% glycerol was used as a carbon source, and it was found that the expression of recombinant interferon-${\alpha}$ could be observed at the beginning of the batch culture when glycerol was used as a carbon source. However, when glucose was used, the initiation of recombinant interferon-${\alpha}$ expression was delayed compared with that when glycerol was used. Furthermore, when the addition of 0.5% glucose was carried out once or twice after 0.5% L-arabinose induction during DO-stat fed-batch culture, the distributions of soluble and insoluble recombinant interferon-${\alpha}$ were modulated. When glucose was not added after the induction of L-arabinose, all of the expressed recombinant interferon-${\alpha}$ formed an inclusion body during the later half of culturing. However, when glucose was added after induction, the expressed recombinant interferon-${\alpha}$ did not all form an inclusion body, and about half of the total recombinant interferon-${\alpha}$ was expressed in a soluble form. It was deduced that the addition of glucose after the induction of L-arabinose might lower the cAMP level, and thus, CAP (catabolite activator protein) might not be activated. The transcription rate of recombinant interferon-${\alpha}$ in the araBAD promoter system might be delayed by the partial repression. This inhibition of the transcription rate probably resulted in more soluble interferon-${\alpha}$ expression caused by the reduction of the protein synthesis rate.

Optimization of Expression Conditions for Soluble Protein by Using a Robotic System of Multi-culture Vessels

  • Ahn, Woo-Sung;Ahn, Ji-Young;Jung, Chan-Hun;Hwang, Kwang-Yeon;Kim, Eunice Eun-Kyeong;Kim, Joon;Im, Ha-Na;Kim, Jin-Oh;Yu, Myeong-Hee;Lee, Cheol-Ju
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1868-1874
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    • 2007
  • We have developed a robotic system for an automated parallel cell cultivation process that enables screening of induction parameters for the soluble expression of recombinant protein. The system is designed for parallelized and simultaneous cultivation of up to 24 different types of cells or a single type of cell at 24 different conditions. Twenty-four culture vessels of about 200 ml are arranged in four columns${\times}$six rows. The system is equipped with four independent thermostated waterbaths, each of which accommodates six culture vessels. A two-channel liquid handler is attached in order to distribute medium from the reservoir to the culture vessels, to transfer seed or other reagents, and to take an aliquot from the growing cells. Cells in each vessel are agitated and aerated by sparging filtered air. We tested the system by growing Escherichia coli BL21(DE3) cells harboring a plasmid for a model protein, and used it in optimizing protein expression conditions by varying the induction temperature and the inducer concentration. The results revealed the usefulness of our custom-made cell cultivation robot in screening optimal conditions for the expression of soluble proteins.

Effect of Soluble EPCR on the Anti-Inflammatory Effects by Activated Protein C (수용성 EPCR에 의한 활성화된 단백질 C의 항염증 작용에 관한 연구)

  • Bae, Jong-Sup;Park, Moon-Ki;Park, Sang-Wook
    • Korean Chemical Engineering Research
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    • v.47 no.4
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    • pp.501-505
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    • 2009
  • In this study, we evaluated the effect of soluble EPCR(Soluble Endothelial Protein C Receptor, sEPCR) on the anti-inflammatory activities by activated protein C(APC) in endothelium. We demonstrated that sEPCR inhibited the barrier protective activity, the inhibition of neutrophils adhesion toward endothelial cells and the inhibition of transendothelial migration by APC in endothelial cells. Interestingly, sEPCR also blocked the mechanism by which APC inhibited the expression of cell adhesion molecules(CAM) by TNF-alpha in endothelial cells. These results suggested that the anti-inflammatory activities of APC was inhibited by sEPCR which blocked the binding motifs of Gla domain of APC to membrane bound EPCR. This finding will provide the important evidence in the development of new medicine for the treatment of severe sepsis and inflammatory diseases and good clue for understanding unknown mechanisms by which APC showed the anti-inflammatory activities in endothelium.

Production of Single-Cell Protein from Starchy Material by the Fusant (전분이용성 세포융합 효모를 이용한 단세포단백질 생산)

  • 정건섭;최신양;구영조;신동화
    • Microbiology and Biotechnology Letters
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    • v.16 no.2
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    • pp.105-110
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    • 1988
  • The production of single cell protein using the amylolytic fusant obtained from cell fusion between Hansenula anomala and Saccharomyces cerevisiae was studied. The fusant12 strain was selected for single cell protein production from starchy materials among five fusants. Optimum nitrogen source and its concentration for the growth of fusant12 were ammonium sulfate and 0.1%, respectively. Optimum concentration of soluble starch and optimum pH of the basal medium were lord and pH 5.6, respectively. Autolysis of fusant12 was effectively carried out by addition of 5% (v/v) ethyl acetate to the cell suspension and liquidization for 30 min before incubation for 24 hr at 3$0^{\circ}C$. Coculture of fusant12 and non-amylolytic yeast, Torulopsis candida YA-l5, resulted in the increase of the mass as compared to the monoculture of fusant12. The cell mass on tapioca medium was increased about 2.5 times as on soluble starch medium. The content of crude protein and nucleic acid of the dry cell were 39% and 5.8%, respectively.

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