• Title/Summary/Keyword: Size exclusion chromatography

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Studies on the Recovery of Triglyceride from Used Shortening by Supercritical Fluid Extraction (초임계유체 추출에 의한 폐식용유의 재활용에 대한 기초연구)

  • Han, Byung-Seok;Yoon, Jung-Ro;Kwon, Young-An;Jung, Mun-Yhung;Kim, Kong-Hwan
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1300-1307
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    • 1999
  • Supercritical fluid extraction was applied to recycling triglyceride from used shortening. Used shortening and its fractions were analyzed with high performance size exclusion chromatography for their composition in triglycerides, polymer and low molecular weight compounds. Conjugated diene value and color of the fractions were also measured with a UV spectrophotometer and a colorimeter, respectively. Pressure and temperature ranges employed were $15{\sim}30$ MPa and $40{\sim}60^{\circ}C$, respectively. Concentration of fat in supercritical (SC) $CO_2$ ranged from $0.3\;X\;10^{-3}{\sim}7.4\;X10^{-3}(g\;fat/g\;CO_2)$. An exponential relation between concentration of fat in SC $CO_2$ and density was observed. Color of the extracts was light yellow which was very close to that of the fresh shortening. Low molecular weight compounds were preferentially concentrated in the initial fraction, while polymer was extracted in the final fraction. Conjugated diene value of the initial fractions was clearly lower than that of feed. It increased sharply as the polymer content in the fraction became significantly large.

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Synthesis and Lubricant Properties of Estolides based on Fatty Acid (지방산 기반 에스토라이드 합성 및 윤활특성)

  • Son, Jeong-Mae;Yoo, Seunghyun;Lee, Sangjun;Shin, Jihoon;Chung, Kunwo;Yang, Youngdo;Kim, Young-Wun
    • Tribology and Lubricants
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    • v.30 no.5
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    • pp.256-264
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    • 2014
  • Enhancing the value of fine chemicals based on biomass resources is an important objective for addressing environmental and other concerns such as demand for renewable or green products, as well as from the political perspective to reduce dependence on fossil feedstock associated with the use of petroleum-based products. Based on these considerations, we studied the synthesis of estolide using waste plant-based oil materials and their application as lubricants and pour point depressants. Five estolides were prepared by varying molar ratio of palmitic acid (PA) to oleic acid (OA) using a reaction time of 48 h. The estolides were characterized by size exclusion chromatography (SEC) and nuclear magnetic resonance (NMR). The isolated yields were in the range of 57-78 % and purity was 93-97%, showing iodine values of 18.2-37.8, total acid numbers (TANs) of 75.6-94.2 mg KOH/g and estolide numbers (ENs) of 1.2-1.8. Increasing the ratio of OA to PA in the synthesis decreased the kinematic viscosity and clouding point of the estolides. Four ball wear test of the estolides as a base oil demonstrated that the wear scar diameter (WSD) of the estolides was significantly lower (0.320-0.495 mm) than the WSD of general base oils such as 150N and Yubase (0.735 and 0.810 mm, respectively), indicating better wear resistance of the estolides. However, the lubricant property was found to be independent of the amount of OA in the estolides. These new materials are prospective candidates for application as a lubricant base oil.

Rheological Characteristics and Molecular Weight of Ammonium-Sulfate Fractions of Tara Gum (염석법에 의한 타라검 분획들의 분자량 및 리올로지 특성)

  • Kim, Kyeong-Yee
    • Korean Journal of Food Science and Technology
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    • v.47 no.3
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    • pp.293-298
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    • 2015
  • This study aimed at characterizing the rheological properties and molecular weight of tara gum fractionated with ammonium sulfate. Tara gum was separated into six fractions (F1-F6) at different concentrations of ammonium sulfate, ranging from 12.21 to 28.67% (w/w). The yield of the tara gum fractions ranged between 4.98 and 17.47%, and their intrinsic viscosity ranged from 9.38 to 12.44 dL/g. The highest values of Huggins coefficient (k') and viscosity-molecular mass were observed in fraction F3. The shear viscosity of the tara gum fractions was measured by a cone-plate viscometer, clearly showing shear thinning behavior. Size-exclusion chromatography results showed that the molecular weight ranged between 635.42 and 776.71 kg/mol, and the F3 fraction exhibited higher values of molecular weight.

Structural characteristics of a red ginseng acidic polysaccharide rhamnogalacturonan I with immunostimulating activity from red ginseng

  • Lee, Sue Jung;In, Gyo;Han, Sung-Tai;Lee, Mi-Hyang;Lee, Jong-Won;Shin, Kwang-Soon
    • Journal of Ginseng Research
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    • v.44 no.4
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    • pp.570-579
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    • 2020
  • Background: Many researchers reported that the various immune activities of red ginseng are due to acid polysaccharides. But, the exact structural characteristics of the acidic polysaccharide in red ginseng have not been fully elucidated. Therefore, we isolated the acidic polysaccharide from red ginseng and characterized the structural property of the active moiety of this polysaccharide, which contributes to the immunostimulatory activity of red ginseng. Methods: A polysaccharide (RGP-AP-I) was purified from red ginseng via size-exclusion chromatography using Sephadex G-100. Immunostimulatary activity of RGP-AP-I was investigated via anti-complementory and macrophage stimulatory activity. The structure of RGP-AP-I was characterized by HPLC, sugar composition, β-glucosyl Yariv reagent and methylation analysis. Results: Peritoneal macrophages stimulated using RGP-AP-I significantly augmented the production of various cytokines such as interleukin (IL)-6, IL-12, and tumor necrosis factor (TNF)-α. The primary structure of RGP-AP-I was elucidated by assessing its sugar composition and methylation analysis. RGP-AP-I is a 96 kDa acidic polysaccharide, and comprises nine different monosaccharides, which mainly include sugars such as rhamnose (Rha, 9.5%), galacturonic acid (GalA, 18.4%), galactose (Gal, 30.4%), and arabinose (Ara, 35.0%). RGP-AP-I exhibited an considerable reaction with the β-glucosyl Yariv reagent, revealing the presence of arabino-β-3,6-galactan. Methylation analysis indicated that RGP-AP-I comprises 21 different glycosyl linkages, such as 3-, 4-, 6- and 3,6-linked Galp; 5-linked Araf; 2,4-linked Rhap; and 4-linked GalAp, which are characteristics of rhamnogalacturonan I (RG-I). Conclusion: we assumed that the immunostimulatory activity of RGP-AP-I may be due to the RG-I structure, which comprises a main chain with a repeating linkage unit, [→2)-Rhap-(1→4)-GalAp-(1→] and three groups of side chains such as (1→5)-linked arabinan, (1→4)-linked galactan, and arabino-β-3,6-galactan, which branch at the C(O)4 positions of Rha residues in the main chain of RGP-AP-I.

Reduction of Antigenicity of Bovine Casein by Microbial Enzymes (미생물효소에 의한 우유 casein의 항원성 저감화)

  • Choe, Hyeon-Seok;Ahn, Jong-Nam;Jeong, Seok-Geun;Ham, Jun-Sang;In, Yeong-Min;Kim, Dong-Un
    • Journal of Dairy Science and Biotechnology
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    • v.21 no.2
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    • pp.97-104
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    • 2003
  • It is extremely important to destroy the antigenicity of milk proteins for dietetic treatment of infants with milk allergy. Enzymatic digestion of milk protein is not only effective for destroying antigenicity, but it also is less liable to alter the nutritive value. Bovine casein was hydrolyzed with eight different commercial proteases derived from bacterias or fungi, either individually or in combination to eliminate protein allergenicity. The average molecular weight of casein hyrdolysates determined by size exclusion chromatography is about 550${\sim}$2,300 dalton range. Antigenicity of the casein hyrdolysates was not detected by heterologous passive cutaneous anaphylaxis in guinea pig-rabbit antiserum system. The inhibition test on the enzyme-linked immunosorbent assay(ELISA) showed that the antigenicity of casein hydrolysates is lowed up to 1/8,000 than that of intact bovine casein. As the enzyme reaction was carried out by the combination of bacterial and fungal protease, casein hydrolysates showed much lower bitterness and antigenicity. It suggests that these hydrolysates will be applied to many kinds of foods including the development of hypo-allergenic infant formula.

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Purification and Characterization of Urushiol Induced Laccase Isoenzyme from Fomitella fraxinea (Urushiol에 의해 유도된 장수버섯 laccase isoenzyme의 정제 및 특성)

  • Choi, Han-Seok;Park, Hyo-Suk;Yeo, Soo-Hwan;Jeong, Seok-Tae;Choi, Ji-Ho;Kim, Myung-Kon
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.152-159
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    • 2010
  • The influence of urushiol, as an allergen on laccase property of Fomitella fraxinea was investigated. The enzyme production was reached to the highest level after 10 days, cultivation and the activity and mycelial biomass were increased by 2.5 and 1.5 folds, respectively, by adding urushiol in the culture medium. In liquid cultures using a Cu Mn-free medium, laccase lactivity was decreased by 3.8-9.2 folds, with similar dry cell weight. Two isoenzymes, were purified using anion exchange, hydrophobic interaction and size-exclusion chromatographies. Both isoenzymes are monomeric proteins, with $M_W$ around 67 kDa(Lac1) and 66 kDa(Lac2), and isoelectric points of 3.67 and 3.81. The optimal conditions for purified isoenzymes were found to be pH 4.5-5.0 and $30-35^{\circ}C$. Activity decreased by the addition of $Fe^{2+}$, $Mg^{2+}$, $Na^+$, and strongly inhibited by EDTA and sodium azide.

Heterologous Expression of Interferon α-2b in Lactococcus lactis and its Biological Activity against Colorectal Cancer Cells

  • Meilina, Lita;Budiarti, Sri;Mustopa, Apon Zaenal;Darusman, Huda Shalahudin;Triratna, Lita;Nugraha, Muhammad Ajietuta;Bilhaq, Muhammad Sabiq;Ningrum, Ratih Asmana
    • Microbiology and Biotechnology Letters
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    • v.49 no.1
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    • pp.75-87
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    • 2021
  • Type I Interferons (IFNα) are known for their role as biological anticancer agents owing to their cell-apoptosis inducing properties. Development of an appropriate, cost-effective host expression system is crucial for meeting the increasing demand for proteins. Therefore, this study aims to develop codon-optimized IFNα-2b in L. lactis NZ3900. These cells express extracellular protein using the NICE system and Usp45 signal peptide. To validate the mature form of the expressed protein, the recombinant IFNα-2b was screened in a human colorectal cancer cell line using the cytotoxicity assay. The IFNα-2b was successfully cloned into the pNZ8148 vector, thereby generating recombinant L. lactis pNZ8148-SPUsp45-IFNα-2b. The computational analysis of codon-optimized IFNα-2b revealed no mutation and amino acid changes; additionally, the codon-optimized IFNα-2b showed 100% similarity with native human IFNα-2b, in the BLAST analysis. The partial size exclusion chromatography (SEC) of extracellular protein yielded a 19 kDa protein, which was further confirmed by its positive binding to anti-IFNα-2b in the western blot analysis. The crude protein and SEC-purified partial fraction showed IC50 values of 33.22 ㎍/ml and 127.2 ㎍/ml, respectively, which indicated better activity than the metabolites of L. lactis NZ3900 (231.8 ㎍/ml). These values were also comparable with those of the regular anticancer drug tamoxifen (105.5 ㎍/ml). These results demonstrated L. lactis as a promising host system that functions by utilizing the pNZ8148 NICE system. Meanwhile, codon-optimized usage of the inserted gene increased the optimal protein expression levels, which could be beneficial for its large-scale production. Taken together, the recombinant L. lactis IFNα-2b is a potential alternative treatment for colorectal cancer. Furthermore, its activity was analyzed in the WiDr cell line, to assess its colorectal anticancer activities in vivo.

Anti-Inflammatory Active Polysaccharide from Postbiotics of Cordyceps militaris Mycelium-Liquid Culture (동충하초(Cordyceps militaris) 균사체 액체발효 포스트바이오틱스로부터 항염증 활성다당 분리)

  • Yeon Suk Kim;Hyun Young Shin;Hoon Kim;Eun-Jin Jeong;Hyun-Gyeong Kim;Min Geun Suh;Hyung Joo Suh;Kwang-Won Yu
    • The Korean Journal of Food And Nutrition
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    • v.36 no.1
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    • pp.6-16
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    • 2023
  • To investigate the anti-inflammatory activity of submerged culture using Cordyceps militaris mycelium, culture-including mycelia was extracted and lyophilized into postbiotics (hot-water extract; CM-HW). HW was fractionated into crude polysaccharide (CM-CP) by ethanol precipitation, and CM-CP was further dialyzed into CM-DCP by dialysis with running water using 12~14 kDa dialysis tube. When the cytotoxicity of subfractions against cells was assessed, no subfraction had a cytotoxic impact that was substantially different from the control groups. In an inflammatory model using LPS-stimulated RAW 264.7 cells, CM-DCP significantly decreased IL-6 and MCP-1 production levels compared to the LPS-control group. CM-DCP also inhibited IL-6 and IL-8 secretion in HaCaT keratinocytes stimulated with TNF-α and IFN-γ. In the meanwhile, the neutral sugar content and mannose ratio of anti-inflammatory CM-DCP were higher than the other fractions, and CM-DCP contained β-1,3/1,6-glucan of 216.1 mg/g. High pressure size exclusion chromatography revealed that CM-DCP contained molecules with a molecular weight range of 5.6 to 144.0 kDa. In conclusion, postbiotics of C. militaris mycelium significantly promoted anti-inflammatory activity, suggesting that neutral polysaccharides including Glc and Man contribute to the anti-inflammation in RAW 264.7 or HaCaT cells.

Production and Evaluation of Immunoreactivity of Poly Lysine-Tagged Single Chain Fragment Variable (ScFv) Lym-1 Antibody for Direct Conjugation to Fluorescence Dye (형광 물질 직접 표지를 위한 Poly Lysine 도입 Lym-1 단일사슬 항체의 제조 및 면역반응성 평가)

  • Jung, Jae-Ho;Choi, Tae-Hyun;Woo, Kwang-Sun;Chung, Wee-Sup;Kang, Joo-Hyun;Jeong, Su-Young;Choi, Chang-Woon;Lim, Sang-Moo;Cheon, Gi-Jeong
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.5
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    • pp.487-494
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    • 2009
  • Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.

The Study on Absorption of $Cd^{2+}$ and $Pb^{2+}$ by Persicaria thunbergii and Rumex crispus, and Their Phytochelatin (고마리와 소리쟁이에 의한 $Cd^{2+}$$Pb^{2+}$의 흡수 및 phytochelatin에 관한 연구)

  • 강경홍;김인성;구정숙
    • Korean Journal of Environmental Biology
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    • v.18 no.2
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    • pp.269-277
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    • 2000
  • When Persicaria thunbergii and Rumex crispus were treated with Cd($NO_3$)$_2$ and Pb($NO_3$)$_2$ of 5 or 10 mM for 5 days, the amount of bioaccumulation of $Pb^{2+}$ in the leaf of P. thunbergii was 2.87-8.08$\mu\textrm{g}$/g and that of $Cd^{2+}$ was 0.82-2.79$\mu\textrm{g}$/g. In the case of P. thunbergii, the concentration of $Pb^{2+}$ in the leaf was higher than that of $Cd^{2+}$. On the other hand, in R. crispus, the concentration of $Cd^{2+}$ and $Pb^{2+}$ were similar as follows ; 1.49$\mu\textrm{g}$/g in $Cd^{2+}$ 5mM, 2.90$\mu\textrm{g}$/g in Cd2+ 10mM, 1.83$\mu\textrm{g}$/g in $Pb^{2+}$ 5mM and 2.73$\mu\textrm{g}$/g in $Pb^{2+}$ 10mM. The remaining rate of heavy metals and the variation of pH in the cultured soil decreased as compared with control (100 % and pH 6.48) after 5 days as follows; to 77.l% and pH 6.39 in $Cd^{2+}$ 5mM, 90.2% and pH 5.79 in $Cd^{2+}$ 10 mM, 81.1% and pH 6.00 in $Pb^{2+}$ 5mM, and 85.7% and pH 5.80 in $Pb^{2+}$ 10 mM. The result of size exclusion chromatography, several phytochelatins were seperated from the extract of the leaf of both plants treated with heavy metals. The molecular mass of these phytochelatins were estimated as follows; in the case of P. thunbergii, about 4,300-8,600 da by $Cd^{2+}$ and about 3,200-9,700 da by $Pb^{2+}$, and in R. crispus, about 4,300 da by $Cd^{2+}$ and about 3,200-7,500 da by $Pb^{2+}$. In addition, $A_{254}$ of these phytochelatins were higher than $A_{280}$. [Phytochelatin, Persicaria thunbergii, Rumex crispus]

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