• Title/Summary/Keyword: Bovine serum albumin

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Protein Partition in an Aqueous Poly (ethyleneglycol)-salt Two-phase System (Poly(ethyleneglycol)과 인산염용액이 형성하는 2상계에서의 단백질 분획에 관한 연구)

  • Lee, Sam-Pin;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
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    • v.19 no.4
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    • pp.285-289
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    • 1987
  • The partition behavior or proteins in an aqueous two-phase system of poly (ethyleneglycol)-potassium phosphate buffer (PEG/PPB) was investigated. The proteins of different surface hydrophobicity, i.e. Bovine serum albumin (BSA), ${\beta}-lactoglobulin$, ovalbumin. moved to the PPB-rich bottom phase in a PEG(12%)/PPB (12%) two-phase system resulting in very low partition coefficients. When the concentration of PPB increased to 15% level. the electric potential of bottom phase changed from +50 mV to zero and the partition coefficient tended to increase. The change In the molar ratio of $K_2HPO_4/KH_2PO_4$ in PPB from 1.43 to 9.55 caused the volume ratio of top to bottom phase $(V_t/V_b)$ to be decreased and protein partition coefficient increased. When the concentration of PPB was elevated from 14% to 26%, the $V_t/V_b$ decreased from 1.5 to 0.39 and the partition coefficient of proteins increased drastically; ${\beta}-lactoglobulin$ 74 fold. BSA 32 fold, ovalbumin 12 fold and lysozyme 5 fold.

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An Enzyme-Linked Immunosorbent Assay for Detection of Pantothenic Acid (판토텐산의 분석을 위한 효소면역측정법)

  • Shon, Dong-Hwa;Park, Youn-Sick;Bae, Gun-Won
    • Korean Journal of Food Science and Technology
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    • v.32 no.5
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    • pp.1009-1014
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    • 2000
  • In order to detect pantothenic acid (PA), conditions for enzyme-linked immunosorbent assay (ELISA) were established. Anti-PA-BSA antibody was produced from rabbits immunized with PA-bovine serum albumin (BSA) conjugates which were prepared by the bromoacetyl chloride [Bc] method (PA-BSA[Bc]) and by the periodate oxidation [Po] method (PA-BSA[Po]). PA-BSA[Bc] and PA-BSA[Po] was used as a coating antigen for competitive indirect(ci)ELISA. The Anti-PA-BSA[Po] antibody on ciELISA showed no competitive reaction. The detection limit of PA by ciELISA using Anti-PA-BSA[Bc] antibody was 1 ppm. The Anti-PA-BSA[Bc] antibody showed little cross-reactivity to PA derivatives such as pantoyllactone, pantetheine, pantothenyl alcohol, and acetyl CoA. The detection limit of PA by microbiological assay (MBA) was 10 ppb. Assay recoveries of PA in egg, cow's liver, and lettuce by ciELISA were 109, 64, and 344%, respectively, comparing with the MBA results.

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Microplate-Based Oxygen Radical Absorbance Capacity (ORAC) Assay of Hydrophilic and Lipophilic Compartments in Plasma

  • Kwak Ho Kyung;Blumberg Jeffrey B.;Chen Chung Yen;Milbury Paul E.
    • Nutritional Sciences
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    • v.9 no.1
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    • pp.48-54
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    • 2006
  • Methods have been developed to evaluate the total antioxidant capacity of foods and plasma but limitations are associated with their ability to determine precisely the contribution of lipophilic antioxidants in a lipid milieu as well as interactions among them Thus, we modified the Oxygen Radical Absorbance Capacity (ORAC) assay to determine the peroxyradical scavenging ability of both hydrophilic and lipophilic compartments in plasma The hydrophilic ORAC assay was performed in a phosphate buffer system utilizing 2,2'-azobis (2-amidinopropane) dihydrochloride as a peroxyradical generator and fluorescein as the target The lipophilic ORAC assay was carried out in a dimethylsulfoxide :butyronitrile (DMSO/BN, 9:1 v/v) system using 2,2'-azobis (2,4-dimethyl valeronitrile) as a peroxyradical generator and BODIPY C11 581/591 as the target Analyses were conducted in bovine serum supplemented with water - and lipid - soluble antioxidants and in human plasma. Albumin (0.5$\sim$5 g/dL) and uric acid (0.1$\sim$0.5 $\mu$mol/L) increased hydrophilic ORAC values in a dose-dependent fashion ($R^{2}$=0.97 and 0.98, respectively) but had no impact on lipophilic ORAC values. $\alpha$-Tocopherol (15$\sim$200 $\mu$mol/L) increased lipophilic ORAC values in a dose-dependent fashion ($R^{2}$=0.94); neither $\alpha$-tocopherol nor $\beta$-carotene had an impact on hydrophilic ORAC values. However, addition of $\beta$-carotene at physiological concentration (0.23$\sim$1.86 $\mu$mol/L), either alone or in combination with other carotenoids, had no significant impact on lipophilic ORAC values. Thus, while assays of 'total antioxidant capacity' in biological matrices would be a useful research and clinical tool, existing methods are limited by the lack of complete responsiveness to the full range of dietary antioxidants.

A Rapid and Universal Direct PCR Method for Macrofungi

  • Park, Mi-Jeong;Lee, Hyorim;Ryoo, Rhim;Jang, Yeongseon;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.49 no.4
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    • pp.455-467
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    • 2021
  • Macrofungi are valuable resources as novel drug candidates, new biomaterials, and edible materials. Recently, genetic approaches pertaining to macrofungi have been continuously growing for their identification, molecular breeding, and genetic engineering. However, purification and amplification of fungal DNA is challenging because of the rigid cell wall and presence of PCR inhibitory metabolites. Here, we established a direct PCR method to provide a rapid and efficient method for PCR-grade macrofungal DNA preparation applicable to both conventional PCR and real-time PCR. We first optimized the procedure of lysis and PCR using the mycelia of Lentinula edodes, one of the most widely consumed macrofungal species. Lysates prepared by neutralizing with (NH4)2SO4 after heating the mycelia in a mixture of TE buffer and KOH at 65℃ for 10 min showed successful amplification in both conventional and real-time PCR. Moreover, the addition of bovine serum albumin to the PCR mixture enhanced the amplification in conventional PCR. Using this method, we successfully amplified not only internal transcribed spacer fragments but also low-copy genes ranging in length from 500 to 3,000 bp. Next, we applied this method to 62 different species (54 genera) of macrofungi, including edible mushrooms, such as Pleurotus ostreatus, and medicinal mushrooms such as Cordyceps militaris. It was found that our method is widely applicable to both ascomycetes and basidiomycetes. We expect that our method will contribute to accelerating PCR-based approaches, such as molecular identification, DNA marker typing, gene cloning, and transformant screening, in macrofungal studies.

Inhibitory Activity of Advanced Glycation Endproducts (AGEs) Formation and Antioxidant Activity of Processed Korean Medicines (포제한약재의 최종당화산물 생성 억제 활성 및 항산화 효과)

  • Lee, AhReum;Kwon, OJun;Choi, JoonYoung;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.32 no.3
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    • pp.63-69
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    • 2017
  • Objectives : Advanced glycation end product (AGEs) is combine formation of glucose and protein. AGEs and reactive oxygen species are potential therapeutic targets for the various disease such as diabetic complications, renal injury, skin damage. The aim of this study was investigated the AGEs inhibitory activity and antioxidant activity of water extracts from 40 Korean medicines and 5 heating-processed Korean medicines. Methods: AGEs formation inhibitory activities of Korean medicines measured using bovine serum albumin (BSA), glucose, and fructose. Then, five effective Korean medicines were selected and heated with 30% ethanol. The AGEs inhibitory activities of heated Korean medicine were measured compared with not-heated Korean medicines. The antioxidant activities were evaluated through radical scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radicals. Furthermore, we examined total phenol and flavonoids contents. Results: Scutellariae Radix, Corni Fructus, Persimmon Fruit, Paeoniae Radix, Mori Folium respectively reduced AGEs production. Morever, heating-processed Scutellariae Radix has AGEs inhibitory activities better than not-processed Scutellariae Radix. Heating- processed Scutellariae Radix scavenged DPPH and ABTS effectively and $IC_{50}$ of DPPH and ABTS radical scavenging activity of Heat processed Scutellariae Radix were $15.47{\pm}0.26{\mu}g/m{\ell}$ and $12.07{\pm}1.23{\mu}g/m{\ell}$. It caused heat processing methods of Scutellariae Radix up regulated total phenol and flavonoids contents ($26.68{\pm}0.01$ to $46.15{\pm}0.10$, $20.30{\pm}0.38$ to $64.20{\pm}0.52$). Conclusion: It has AGEs inhibitory activities that 20 kind of medicinal plants of 40 medicinal plants. Especially, heat processed Scutellariae Radix has excellent AGEs inhibitory activities and antioxidant effect.

High fat diet-induced brain damaging effects through autophagy-mediated senescence, inflammation and apoptosis mitigated by ginsenoside F1-enhanced mixture

  • Hou, Jingang;Jeon, Byeongmin;Baek, Jongin;Yun, Yeejin;Kim, Daeun;Chang, Boyoon;Kim, Sungyeon;Kim, Sunchang
    • Journal of Ginseng Research
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    • v.46 no.1
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    • pp.79-90
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    • 2022
  • Background: Herbal medicines are popular approaches to capably prevent and treat obesity and its related diseases. Excessive exposure to dietary lipids causes oxidative stress and inflammation, which possibly induces cellular senescence and contribute the damaging effects in brain. The potential roles of selective enhanced ginsenoside in regulating high fat diet (HFD)-induced brain damage remain unknown. Methods: The protection function of Ginsenoside F1-enhanced mixture (SGB121) was evaluated by in vivo and in vitro experiments. Human primary astrocytes and SH-SY5Y cells were treated with palmitic acid conjugated Bovine Serum Albumin, and the effects of SGB121 were determined by MTT and lipid uptake assays. For in vivo tests, C57BL/6J mice were fed with high fat diet for 3 months with or without SGB121 administration. Thereafter, immunohistochemistry, western blot, PCR and ELISA assays were conducted with brain tissues. Results and conclusion: SGB121 selectively suppressed HFD-induced oxidative stress and cellular senescence in brain, and reduced subsequent inflammation responses manifested by abrogated secretion of IL-6, IL-1β and TNFα via NF-κB signaling pathway. Interestingly, SGB121 protects against HFD-induced damage by improving mitophagy and endoplasmic reticulum-stress associated autophagy flux and inhibiting apoptosis. In addition, SGB121 regulates lipid uptake and accumulation by FATP4 and PPARα. SGB121 significantly abates excessively phosphorylated tau protein in the cortex and GFAP activation in corpus callosum. Together, our results suggest that SGB121 is able to favor the resistance of brain to HFD-induced damage, therefore provide explicit evidence of the potential to be a functional food.

A Study on the Physicochemical Properties and Antioxidative Activity of Whey Protein Isolate (WPI의 이화학적 특성과 항산화성에 관한 연구)

  • Ahn, Myung-Soo;Kim, Chan-Hee
    • Journal of the Korean Society of Food Culture
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    • v.22 no.1
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    • pp.97-103
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    • 2007
  • In this study, physicochemical properties and the antioxidative activity of whey protein isolate(WPI) for com germ oil were measured. The pH of WPI was 6.26, and the titrable acidity was 0.18%. The WPI’s moisture content was 5.2% and each of the other element content such as lactose, crude protein, crude ash and crude fat was found to be 0.8%, 90.7%, 2.7% and 0.6%, respectively. The amounts of active SH group in WPI 9 ${\mu}$ M-g and total colony counts of bacteria was 5.9 ${\times}$ 10$^3$ CFU-g. ${\alpha}$-Lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin(BSA) were shown in WPI as major protein by electrophoresis. The antioxidative effect of WPI and other antioxidants on com germ oil used as substrate was determined by peroxide value(POV) and conjuqated dienoic acid value(CDV). By these results, the order of antioxidative effects could be defined as BHT 0.02%>ascorbic acid 0.1%>WPI 0.1%>WPI 0.02%>ascorbic acid 0.02%>control>tocopherol 0.02%>tocopherol 0.1%, respectively. Also the induction period of com germ oil added with WPI was longer by 1.6 times than that of control(none added any antioxidant). Therefore the fact suggested that WPI could be utilized as a good antioxidative agents.

Preparation of Protein-coated Cationic Liposomes Containing Doxorubicin and Their Binding Property of Blood Plasma Protein (독소루비신을 함유하고 단백질로 수식된 양이온성 리포솜의 제조 및 혈장 단백흡착 특성)

  • Kim, Sung-Kyu;Jung, Soon-Hwa;Jung, Suk-Hyun;Seong, Ha-Soo;Chi, Sang-Cheol;Cho, Sun-Hang;Shin, Byung-Cheol
    • Journal of the Korean Chemical Society
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    • v.52 no.1
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    • pp.57-65
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    • 2008
  • are nanometer or micrometer scale vesicles that can be used as drug delivery carriers. However, plain liposomes are plagued by rapid opsonization, making their circulation time in bloodstream be shortened. In this study, model protein, bovine serum albumin (BSA)-coated liposomes were prepared by coating cationic liposomes with BSA molecules at higher pH than isoelectric point of BSA. The BSA molecules coated on the liposomal surface were denatured by thermal treatment at above 60oC. While both plain and cationic liposomes had about mean particle diameter of 1041 nm, BSA-coated cationic liposomes (BCL) had mean particle diameter of 1091 nm. Encapsulation of model drug, doxorubicin (DOX), in liposomes were carried out by using remote loading method and the loading efficiency of DOX to liposomes was about 90%. The mean particle diameter of BCL did not increase in blood plasma and adsorption of plasma protein was much less than plain or cationic liposomes. These results suggest that BCL can be used as a long-circulating liposomes in bloodstream.

The Release of Albumin from PLGA and PCL Wafers Containing Natural and Synthetic Additives for Protein Delivery (단백질 전달체로서 천연 및 합성재료의 첨가에 따른 PLGA와 PCL웨이퍼로부터 알부민의 방출거동)

  • Hyun Hoon;Lee Jae Ho;Seo Kwang Su;Kim Moon Suk;Rhee Jhon M.;Lee Hai Bang;Khang Gilson
    • Polymer(Korea)
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    • v.29 no.5
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    • pp.468-474
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    • 2005
  • PLGA and PCL copolymers initiated by carbitol as drug carriers were synthesized by ring-opening polymerization of L-lactide (LA), glycolide (GA), and $\varepsilon-caprolactone(\varepsilon-CL)$. Implantable wafers were simply fabricated by direct compression method after physical mixing of copolymers and bovine serum albumin-fluorescein isothiocyanate (BSA-FITC) as a model protein drug. The release amounts of BSA-FITC from wafers were determined by fluorescence intensity using the fluorescence spectrophotometer. Also, the release behavior of BSA-FITC on wafers was controlled by adding the additives such as collagen, small intestinal submucosa (SIS), poly(vinyl pyrrolidone) (PVP), and poly(thylene glycol) (PEG). The wafer prepared by PLGA and PCL exhibited slow release within $10\%$ for 30 days. But, those prepared by a variety of additives exhibited the controlled BSA release patterns with a dependence on the additive contents. furthermore, the wafers containing natural materials such as collagen and SIS showed more zero-order release profile than that with synthetic materials such as PVP and PEG. It was confirmed that the release of BSA from implantable wafers could be easily controlled by adding natural additives.

Skin Wrinkle Improvement Effect of Paeoniae radix and processed Paeoniae radix Through inhibition of Advanced glycation end products (AGEs) (작약 및 포제작약의 최종당화산물 억제를 통한 피부 주름 개선 효과)

  • Kim, SuJi;Lee, AhReum;Kim, SooHyun;Kim, KyeongJo;Kwon, OJun;Choi, JoonYoung;Koo, JinSuk;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.32 no.4
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    • pp.53-60
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    • 2017
  • Objectives : Collagen decrease of Skin appears through various path ways. One of causes may be the Advanced glycation endproducts (AGEs) that combine formation of glucose and protein. The aim of this study was to explore the prevent wrinkle formation of Paeoniae radix (PR) and heated Paeoniae radix (HPR) via AGEs path way. Methods : AGEs formation inhibitory activities of PR and HPR measured using bovine serum albumin, glucose, and fructose. To evaluate the protective effects of PR and HPR in diabetic rats induced with streptozotocin (STZ) and methyl glyoxal (MGO), SD rat were distributed into four groups. Normal rats (Nor), AGEs-induced rats (Con), AGEs-induced rats treated with 100 mg/kg PR(PR), AGEs-induced rats treated with 100 mg/kg HPR (HPR). To induce AGEs, streptozotocin (50 mg/kg) was administered intraperitoneally and after 3 days administrated 100mM methyl glyoxal for 3 weeks. Results : The oral administration of HPR inhibited AGEs in skin tissues compared with PR. The increased reactive oxygen species (ROS) levels in the serum were diminished by HPR treatment. The analyses of kidney and skin tissues proteins indicated that HPR treatment effectively reduced AGEs related protein levels as compared to that by PR treatment. Also, HPR decreased anti-oxidant related protein levels in skin tissues such as catalase, glutathione peroxidase. Moreover, it inhibited the reduction of COL1A2 by decreasing MMP-1. Conclusion : Based on these results, it was suggested that PR and HPR could have Improving effects on wrinkle formation. These evidences provide useful information for the development wrinkle formation treated agent.