• Title/Summary/Keyword: bovine serum albumin

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Experimental Study on Effects of Cucurbita moschata Duch. on Antioxidation (호박잎의 항산화 효과 연구)

  • Cha, Yun-Yeop
    • Journal of Korean Medicine for Obesity Research
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    • v.9 no.2
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    • pp.57-63
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    • 2009
  • In recent year, We are concerned about anti-aging, disease-protection, long-life, many methods are used in solving this problem. And Those are related with antioxidative ability. Recently, We heard that Cucurbita moschata Duch. has anti-hypertensive effect and good for health. So I let made a experiment for this result of the anti-oxidant effect of Cucurbita moschata Duch. used for 3 methods, those are DPPH radical scavenging activity, Nitric oxide(NO) radical scavenging activity and Bovine serum albumin(BSA). The results of this study were as follows: We measured level of DPPH radical scavenging activity. And we obtained results that the ability of DPPH radical's elimination was increased when concentration of Cucurbita moschata Duch. was to $20\;mg/m{\ell}$. We measured level of Nitric oxide(NO) radical scavenging activity. And we founded that the ability of NO radical's elimination was increased when concentration of Cucurbita moschata Duch. was to $10\;mg/m{\ell}$. When we inspected antioxidation with Bovine serum albumin(BSA). And we obtained that antioxidative ability was increased after$2.5\;mg/m{\ell}$ of Cucurbita moschata Duch.. So I guess that Cucurbita moschata Duch. has effect of antioxidative ability. Hereafter we need differential experimental methods for evidence of antioxidative effect on Cucurbita moschata Duch.

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The Influence of Bakers' Yeast Cells on Protein Adsorption Performance in Dye-Ligand Expanded Bed Chromatography

  • Chow, Yen Mei;Tey, Beng Ti;Ibrahim, Mohd Nordin;Ariff, Arbakariya;Ling, Tau Chuan
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.552-555
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    • 2005
  • The influence of whole yeast cells $(0{\sim}15%\;w/v)$ on the protein adsorption performance in dye-ligand chromatography was explored. The adsorption of a model protein, bovine serum albumin (BSA), was selected to demonstrate this approach. The UpFront adsorbent $(p=1.5\;g/cm^3)$ derivatised with Cibacron Blue 3GA and a commercially available expanded bed column (20 mm i.d.) from UpFront Chromatography, Denmark, were employed in the batch binding and expanded bed operation. The BSA binding capacity was demonstrated to not be adversely affected by the presence of yeast cells. The dynamic binding capacity of BSA at a $C/C_0=0.1$ biomass concentration of 5, 10, 15% w/v were 9, 8, and 7.5mg/mL of settled adsorbent, respectively.

Protein Adsorption and Hydrodynamic Stability of a Dense, Pellicular Adsorbent in High-Biomass Expanded Bed Chromatography

  • Chow, Yen Mei;Tey, Beng Ti;Ibrahim, Mohd Nordin;Ariff, Arbakariya;Ling, Tae Chuan
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.3
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    • pp.268-272
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    • 2006
  • A dense, pellicular UpFront adsorbent ($p=1.5 g/cm^3$, UpFront Chromatography, Cophenhagen, Denmark) was characterized in terms of hydrodynamic properties and protein adsorption performance in expanded bed chromatography. Cibacron Blue 3GA was immobilised into the adsorbent and protein adsorption of bovine serum albumin (BSA) was selected to test the setup. The Bodenstein number and axial dispersion coefficient estimated for this dense pellicular adsorbent was 54 and $1.63{\times}10^{-5}m^2/s$, respectively, indicating a stable expanded bed. It could be shown that the BSA protein was captured by the adsorbent in the presence of 30% (w/v) of whole-yeast cells with an estimated dynamic binding capacity $(C/C_o = 0.01)$ of approximately 6.5 mg/mL adsorbent.

Subtilisin QK, a Fibrinolytic Enzyme, Inhibits the Exogenous Nitrite and Hydrogen Peroxide Induced Protein Nitration, inVitro and inVivo

  • Ko, Ju-Ho;Yan, Junpeng;Zhu, Lei;Qi, Yipeng
    • BMB Reports
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    • v.38 no.5
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    • pp.577-583
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    • 2005
  • Subtilisin QK, which is newly identified as a fibrinolytic enzyme from Bacillus subtilis QK02, has the ability of preventing nitrotyrosine formation in bovine serum albumin induced by nitrite, hydrogen peroxide and hemoglobin in vitro verified by ELISA, Western-blot and spectrophotometer assay. Subtilisin QK also attenuates the fluorescence emission spectra of bovine serum albumin in the course of oxidation caused by nitrite, hydrogen peroxide and hemoglobin. Furthermore, subtilisin QK could suppress the transformation of oxy-hemoglobin to met-hemoglobin caused by sodium nitrite, but not the heat-treated subtilisn QK. Compared with some other fibrinolytic enzymes and inactivated subtilisin QK treated by phenylmethylsulfonylfluoride, the ability of inhibiting met-hemoglobin formation of subtilisin QK reveals that the anti-oxidative ability of subtilisin QK is not concerned with its fibrinolytic function. Additionally, nitrotyrosine formation in proteins from brain, heart, liver, kidney, and muscle of mice that is intramuscular injected the mixture of nitrite, hydrogen peroxide and hemoglobin is attenuated by subtilisin QK. Subtilisin QK can also protect Human umbilical vein endothelial cell (ECV-304) from the damage caused by nitrite and hydrogen peroxide.

Peroxynitrite Scavenging Activity and its Mechanism of Cheonga-hwan (청아환의 Peroxynitrite 제거 활성 및 기전)

  • 김성호;정지천
    • The Journal of Korean Medicine
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    • v.23 no.4
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    • pp.55-63
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    • 2002
  • Objectives: Peroxynitrite ($ONOO^{-}$), formed from the reaction of superoxide <${\cdot}O_2^{-}$) and nitric oxide (NO), is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been implicated in diseases such as aging process, Alzheimer's disease, rheumatoid arthritis, cancer and arteriosclerosis. Due to the lack of endogenous enzymes responsible for $ONOO^{-}$ inactivation, developing a specific $ONOO^{-}$ scavenger is of considerable importance. The aim of this study was to evaluate $ONOO^{-}$ scavenging activity and its mechanism in Cheonga-hwan (CAH). Methods: The $ONOO^{-}$ scavenging activity in CAH was assayed by measuring oxidized dihydrorhodamine 123 (DHR 123) by fluorescence. The scavenging efficacy was expressed as $IC_{50}$, showing the concentration of each sample required to cause 50% inhibition of DHR 123 oxidation. In a separate study, the protective effect of CAR on $ONOO^{-}$-induced nitration of bovine serum albumin (BSA) was investigated using immunoassay with a monoclonal anti-nitrotyrosine antibody, and a horseradish peroxidase-conjugated anti-mouse secondary antibody from sheep. Results: CAH showed potent scavenging activities of $ONOO^{-}$, NO and ${\cdot}O_2^{-}$. The data demonstrated that CAH led to decreased $ONOO^{-}$-mediated nitration of tyrosine through electron donation. CAH showed significant inhibition on nitration of bovine serum albumin by $ONOO^{-}$ in a dose-dependent manner. Conclusions: CAH can be developed as an effective peroxynitrite scavenger for the prevention of the $ONOO^{-}$ involved diseases.

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Preparation and Characterization of Poly[3-hydroxybutyrate-co-4-hydroxybutyrate] Microsphere (Poly[3-hydroxybutyrate-co-4-hydroxybutyrate] 미립구의 제조 및 특성화)

  • Kang, Hye-Su;Kim, Beom-Soo
    • KSBB Journal
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    • v.22 no.3
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    • pp.146-150
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    • 2007
  • Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] microspheres were prepared using solvent evaporation technique. P(3HB-co-4HB) with 3.9 mol% 4HB was synthesized by fed-batch culture of Ralstonia eutropha. The effects of concentration and type of surfactant (Tween 80, sodium dodecylsulfate, and polyvinyl alcohol), addition of dispersion stabilizer (Acacia), concentration of polymer and model drug (bovine serum albumin) on particle size of the microspheres and their in vitro drug release characteristics were investigated. The average particle size of the microspheres decreased with the addition of dispersion stabilizer and increased with the concentration of surfactant, drug and polymer. Amount of drug release increased with decreasing particle size of the microspheres.

Analysis of the Formation of Protoplasts and Regeneration of Cells in Phycomyces blakesleeanus

  • Joe, Fukui;Choi, Kwan-Sam;Atsushi Miyazaki;Tamotsu Ootaki;Taneaki Oikawa
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.72-78
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    • 2001
  • It is possible ot prepare protoplasts of the zygomycete fungus, Phycomyces blakesleeanus, by digesting the cell wall of spore germlings with commercially available chitinase and chitosanase. However, the cells without any cell walls immediately form large aggregates, and thus, it is difficult to isolate the individually separated protoplasts. Inherent problem with the formation of aggregates in preparing protoplasts could be solved by the use of bovine serum albumin (BSA). As a result, we were able to prepare a large number of single protoplsts quickly and easily. We took time-lapse photomicrographs of the formation of protoplasts, and found that there were certain regions of the cell wall of spore germlings that were sensitive to chitinase and chitosanase, although the cell wall of the original spores is known to be insensitive to these enzymes. There are two kinds of cell walls on a spore germling; one with a bound wheat germ agglutinin (WGA), and the other a bound concanavalin A (ConA). Furthermore, only cells with walls which had bound WGA were able to regenerate, while those with walls with bound ConA were not able to regenerate.

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Effect of Linker for Immobilization of Glutathione on BSA-Assembled Controlled Pore Glass Beads

  • Chen, Li-Hua;Choi, Young-Seo;Park, Jung-Won;Kwon, Joseph;Wang, Rong-Shun;Lee, Tae-Hoon;Ryu, Sung-Ho;Park, Joon-Won
    • Bulletin of the Korean Chemical Society
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    • v.25 no.9
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    • pp.1366-1370
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    • 2004
  • Controlled pore glass bead was modified with bovine serum albumin (BSA), and glutathione (GSH) was immobilized through three kinds of linkers on top of BSA. Bis(3-sulfo-N-hydroxysuccinimide suberate) sodium salt $(BS^3)$, N-hydroxysuccinimide 3-(2-pyridyldithio)propionate (SPDP), or N-hydroxysuccinimide 4-maleimidobutyrate (GMBS) was introduced into the BSA-bound matrix. Subsequently, GSH was immobilized by addition of thiol side chain into the maleimido moiety, replacing a disulfide group, or formation of an amide group upon releasing 3-sulfo-N-hydroxysuccimide group. It was observed that conjugation methodology played a critical role for activity of the immobilized GSH. SDS-PAGE chromatogram showed that the matrix of glutathione immobilized on BSA through GMBS manifested high selectivity towards glutathione-S-transferase (GST) in cell lysate.

Effect of Bovine Serum Albumin on the Stability of Methotrexate-encapsulated Liposomes

  • Kim, Chong-Kook;Kim, Han-Sung;Lee, Beum-Jin;Han, Jeong-Hee
    • Archives of Pharmacal Research
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    • v.14 no.4
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    • pp.336-341
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    • 1991
  • The effect of bovine serum albumin (BSA) on the encapsulation efficiency and stability of liposomes containing methotrexate (MTX) having different surface charges and cholesterol contents were investigated. The encapsulation efficiency of MTX was lower and the release of MTX was faster by the addition of BSA. The leaking of MTX from lipid bilayer depends upon the BSA concentrations. These results may be derived from the interaction of BSA with lipid bilayers. The dynamic structural changes of BSA were monitored indirectly using circular dichroism spectra. Observed dynamic structural changes of BSA with liposomes are presumed to reflect the interaction of BSA with liposomes. Negatively charged liposomes have more strong interaction with BSA than neutral and positively charged liposomes. BSA attacks lipid bilayers whether it is at the inner or at the outer phase of lipid bilayer and induces leakage of entrapped MTX. Especially, negatively charged liposomes are more sensitive than others. The inclusion of cholesterol in the lipid layers inhibits the interaction of BSA with liposomes and shows protective effect against BSA-induced leakage of MTX. To endure the attacking of BSA liposomes as drug carriers should be made using cholesterol.

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Protein-Coating Evaluation Method of Colloidal Gold Nanoparticles (콜로이드 골드 나노입자의 단백질 수송성 평가법)

  • Kim, Mi-Young;Noh, Sang-Myoung;Kim, Jung-Mogg;Choi, Han-Gon;Kim, Jung-Ae;Oh, Yu-Kyoung
    • Journal of Pharmaceutical Investigation
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    • v.34 no.6
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    • pp.465-469
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    • 2004
  • Colloidal gold nanoparticles might be of use as nano scale delivery systems of various therapeutic materials in the future. Recent studies have reported the feasibility of colloidal gold nanoparticles as gene delivery systems or protein delivery systems. In this study, we aimed to develop a short-step method useful for screening the optimal coating conditions of colloidal gold nanoparticles with proteins. We observed that colloidal gold nanoparticles have properties of changing its unique color when they were exposed to NaCl solution. Taking advantage of the color changing properties of colloidal gold nanoparticles, we applied the color testing method of colloidal gold nanoparticles solutions for evaluating the protein coating nature. Using bovine serum albumin as a model protein, we tested the protein coating of colloidal gold nanoparticles via the color change upon NaCl addition. The optimal coating concentration and coating conditions of colloidal gold nanoparticles with bovine serum albumin were fixed using the color testing methods. We suggest that the color testing method might be applied to optimize the coating condition of colloidal gold nanoparticles with other therapeutic proteins.