• Title/Summary/Keyword: gel permeation chromatography

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Debittering of Citrus Products Using ${\beta}-Cyclodextrin$ Polymer and Ultrafiltration Process (${\beta}-Cyclodextrin$ 중합체와 한외여과 공정을 이용한 감귤류의 쓴맛 성분 제거)

  • Woo, Gun-Jo;Ha, Seung-Mi
    • Korean Journal of Food Science and Technology
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    • v.29 no.2
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    • pp.302-308
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    • 1997
  • ${\beta}-Cyclodextrin\;({\beta}-CD)$ polymers were prepared using epichlorohydrin as a cross linking agent. The polymers were separated into ${\beta}-CD$ soluble polymer $({\beta}-CD\;SP)$ and ${\beta}-CD$ insoluble polymer $({\beta}-CD\;ISP)$ on a 10,000 molecular weight cut-off membrane (YM 10). Optimum separation conditions in the YM 10 were: transmembrane pressure 51.7 kPa, separation temperature $35^{\circ}C$, and volume concentration ratio 10. The flux was $0.025\;mL/cm^{2}/min$ under the optimum conditions. Gel permeation chromatography indicated that ${\beta}-CD\;SP\;and\;{\beta}-CD\;ISP$ had a degree of polymerization of $2{\sim}8$ and over 10, respectively. The formation of an inclusion complex with hydrophobic compounds such as 4-dimethylaminoazobenzene, methyl red, and naringin was compared among ${\beta}-CD,\;{\beta}-CD\;SP\;and\;{\beta}-CD\;ISP$. The molar absorptivity for the two chromatic compounds was increased and the absorption peak was shifted in the presence of ${\beta}-CD$ polymers. Naringin, the principal flavonoid bitter tasting component of citrus fruit, had a low water solubility. The solubility of naringin was increased through the formation of an inclusion complex with ${\beta}-CD$ polymers. There was no significant difference in the formation of an inclusion complex between ${\beta}-CD\;SP\;and\;{\beta}-CD\;ISP$. Reduction of the bitter components from citrus products was shown to be possible when employing ${\beta}-CD\;SP$, while the usage of ${\beta}-CD$ monomer has been limited due to the low water solubility.

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Properties of Pectin Extracted from By-product in Citrus Processing (밀감 가공부산물에서 추출한 펙틴의 특성)

  • 박용곤;강윤한;차환수;김흥만;석호문
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.4
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    • pp.659-664
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    • 1996
  • The amount and characteristics of pectin in the albedo and flavedo layers of the citrus peels, and those of the pulp were investigated. Alcohol insoluble solid(AIS) content was the highest in albedo layer(18.1%), and the lowest in pulp(5.7%). The pulp and the albedo layer showed a potential pectin sources as containing pectins of 40.5% and 35.2% of the total polysaccharides of the pulp and the albedo layer, respectively. Total pectin contents were about 30% of the AIS and showed comparatively constant values among the byproducts. Hydrochloric acid soluble pectin contents were the hightest in the flavedo layer, 14.0%, and the lowest in the pulp, 4.4%. Over 90% of the total pectin could be extracted after 60min with 0.05N HCI at $85^{\circ}C.$ Microwave treatment reduced the extraction time significantly ; a comparable extraction yield was acquired after 10min with microwave treatment. The degree of esterification of the extracted pectin also increased with microwave treatment. Neutral sugars in the hydrolysate of the pectin were rhamnose, arabinose, galactose, glucose and xylose. No differences in molecular weight distribution of the pectin were found between the albedo and flavedo layers. Pectin of the pulp showed different molecular weight distribution from that of the peels.

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Evaluation of Permanent Deformation Characteristics of Recycled Asphalt Concretes Made by Improved Binder-Rejuvenation (바인더 회생방식을 개선한 재생 아스팔트 콘크리트의 소성변형 특성 연구)

  • Kim Kwang-Woo;Kweon Oh-Sun;Doh Young-Soo
    • International Journal of Highway Engineering
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    • v.8 no.1 s.27
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    • pp.1-13
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    • 2006
  • This paper is one of the studies for developing new methodologies for improving performance of hot-mix recycled asphalt mixtures. The objective of this study is to evaluate rut-resistance characteristics of recycled asphalt mixture which was prepared by newly developed mixing method. The new mixing method provided more sufficient rejuvenation of old binder of reclaimed asphalt pavement (RAP), making homogeneous binder viscosity level in a recycled mixture. Two aggregates (gneiss and granite), two RAP contents (15% and 30%) and two contents (none and 6%) of polymer modifier (LDPE) were used. Recycled mixture was prepared with two methods; method A and method F. To examine difference of binder oxidation level by type of material within a recycled mixture, Gel-permeation chromatography(GPC) analysis was performed on the binders mixed with coarse aggregates and matrix separately. Laboratory tests were performed for evaluation of rut resistance characteristics of each recycled mixture and these includes wheel tracking (WT) test and Kim test. Rut depth and dynamic stability were obtained from WT test and deformation strength $(S_D)$ was obtained from Kim test. The results of regression analysis was shown that correlation $(R^2)$ of F mixing mixtures was higher than one of A mixing mixtures. Therefore, F mixing mixtures showed more consistent rut resistance than h mixing mixtures.

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Analysis of Binder Aging Levels in Asphalt Mixture Caused by Short-term Aging Conditions Using the GPC Technique (GPC를 이용한 아스팔트 혼합물의 단기노화 조건에 따른 노화도 분석)

  • Kim, Yeongsam;Yun, Jiyeon;Jeong, Seungho;Kim, Kwangwoo
    • International Journal of Highway Engineering
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    • v.18 no.3
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    • pp.1-9
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    • 2016
  • PURPOSES : This study aims to show the difference of the binder aging level in the hot-mix asphalt (HMA) mixture after short-term aging (SA) under different aging conditions, such as mixture temperature and duration in hour. METHODS : Three SA times (i.e., 1 h, 2 h, and 4 h) at two temperatures (i.e., $160^{\circ}C$ and $180^{\circ}C$) were used for the normal mixtures prepared using a PG64-22 asphalt. The field long-term aging (LA) was simulated by applying the same LA procedure (65 h at $110^{\circ}C$) to all compacted specimens, prepared at the air void of 7% using each SA-treated mixture, in a convection oven. The binder aging level was measured in terms of large molecular size by gel-permeation chromatography (GPC) from the mixture and the absolute viscosity (AV) from the recovered binder. The aging levels were evaluated using those two properties after SA and LA, and then compared based on the normal SA (NSA) mixture (1 h at $160^{\circ}C$). The service life reduction caused by SA in various conditions was estimated based on the aging level of the field cores from different locations in various service lives. RESULTS : The results of the laboratory evaluation indicated that the binder of the mixture, which was treated at longer SA time and higher temperature, showed a significantly higher aging level than the NSA mixture. The binder aging level from a longer time, such as 2 h and 4 h SA, or at a higher temperature ($180^{\circ}C$), were estimated to be similar to that of the mixtures, which had already been in field service for several years. CONCLUSIONS : The HMA mixture should be produced at a moderate temperature, such as $160^{\circ}C$, and placed within a limited hauling and queuing time to avoid a significant short-term aging of the binder before placement in the field pavement. The SA for a longer time at a higher temperature than the NSA condition was found to be detrimental to the service life of the asphalt pavement.

Isolation and Characterization of a Novel Polysaccharide Producing Bacillus polymyxa A49 KCTC 4648P

  • Ahn, Sung-Gu;Suh, Hyun-Hyo;Lee, Chang-Ho;Moon, Seong-Hoon;Kim, Hee-Sik;Ahn, Keug-Hyun;Kwon, Gi-Seok;Oh, Hee-Mock;Yoon, Byung-Dae
    • Journal of Microbiology and Biotechnology
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    • v.8 no.2
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    • pp.171-177
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    • 1998
  • The strain A49, which produces a new type of extracellular polysaccharide was isolated from soil samples. From morphological, physiological and biochemical tests, the strain A49 was identified as a Bacillus polymyxa and named Bacillus polymyxa A49. Bacillus polymyxa A49 was found to produce a highly viscous extracellular polysaccharide when grown aerobically in a medium containing glucose as the sole source of carbon. The polysaccharide (A49 POL) showed a homogeneous pattern on gel permeation chromatography (GPC) and its molecular weight was estimated to be about 1.6 mega dalton (mDa). The FT-IR spectrum of A49-POL revealed typical characteristics of polysaccharides. As a result of investigations with HPLC and carbozole assay, A49-POL was found to consist of L-fucose, D-galactose, D-glucose, D-mannose, and D-glucuronic acid, with the molar ratio of these sugars being approximately 1:2:7:50:12. Rheological analysis of A49 POL revealed that it is pseudoplastic and has a higher apparent viscosity at dilute concentrations than does xanthan gum. The consistancy factor of A49 POL was found to be higher, and the flow index of A49 POL lower, than xanthan gum. Its apparent viscosity was comparatively unstable at various temperatures. the A49 POL showed the highest apparent viscosity at pH 3. When salts were added to A49 POL solution, the solution was compatible with up to 10% KCl, 35% NaCl, 55% $CaCl_2$, 55% $MgCl_2$, 55% $K_2HPO_4$, and 110% $Ca({NO_3})_2$, respectively.

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Effects of Microbacterium laevaniformans Levans Molecular Weight on Cytotoxicity

  • Oh, Im-Kyung;Yoo, Sang-Ho;Bae, In-Young;Cha, Jae-Ho;Lee, Hyeon-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.985-990
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    • 2004
  • Levans produced from Microbacterium laevaniformans were isolated, characterized, and fractionated by molecular weight. TLC, HPLC, and GC-MS analyses of the exopolysaccharide showed that it was a fructan-type polymer and was composed of (2,6)- and (2,1)-glycosidic linkages. $^{13}C$-NMR analysis proved that the polysaccharide was mainly a $\beta$-(2,6)-linked levan-type polysaccharide. To investigate the cytotoxicity of the acetone-precipitated levan fractions such as M1, M2, and M3, HepG2, P388D1, U937, SNU-1, and SNUC2A cell lines were screened. Among the cell lines tested, the cytotoxicity of M1- M3 fractions were detected from only SNU-1 and HepG2 cells at the dosage level of $100-800\mu\textrm{g}ml$. The M2 fraction M_r$, 80,000) at 400 $mu{g/ml}$ had the greatest cell growth inhibition (84.6%) on SNU-1, while the M1 $(M_r$, 50,000) at $800\mu\textrm{g}ml$ showed the greatest (46.32%) on HepG2. To obtain more uniform M_r$ fractions of levan, the levan was further fractionated from S1 $(M_r$ 1,000,000) to S5 $(M_r$ 10,000) using gel permeation chromatography. Again, the S1-S5 fractions had strong cytotoxicity on SNU-1 and HepG2 cell lines. The greatest inhibition effects of S4 $(M_r$ 80,000) on SNU-1 and S5 $(M_r$ 10,000) on HepG2 were shown to be 49.5% and 73.0%, respectively. The cytotoxicity of the levan fractions was more effective on SNU-1 than on HepG2. Although the relationship between the Mw and the cytotoxicity was not clear, smaller $M_r$, fractions of levan showed greater growth inhibition effect on the cancer cell lines in general. Therefore, it was indicated that a specific Mw class of levan is responsible for the effective cytotoxicity.

Effects of N-/C-Terminal Extra Tags on the Optimal Reaction Conditions, Activity, and Quaternary Structure of Bacillus thuringiensis Glucose 1-Dehydrogenase

  • Hyun, Jeongwoo;Abigail, Maria;Choo, Jin Woo;Ryu, Jin;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1708-1716
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    • 2016
  • Glucose dehydrogenase (GDH) is an oxidoreductase enzyme and is used as a biocatalyst to regenerate NAD(P)H in reductase-mediated chiral synthesis reactions. In this study, the glucose 1-dehydrogenase B gene (gdhB) was cloned from Bacillus thuringiensis subsp. kurstaki, and wild-type (GDH-BTWT) and His-tagged (GDH-BTN-His, GDH-BTC-His) enzymes were produced in Escherichia coli BL21 (DE3). All enzymes were produced in the soluble forms from E. coli. GDH-BTWT and GDH-BTN-His showed high specific enzymatic activities of 6.6 U/mg and 5.5 U/mg, respectively, whereas GDH-BTC-His showed a very low specific enzymatic activity of 0.020 U/mg. These results suggest that the intact C-terminal carboxyl group is important for GDH-BT activity. GDH-BTWT was stable up to 65℃, whereas GDH-BTN-His and GDH-BTC-His were stable up to 45℃. Gel permeation chromatography showed that GDH-BTWT is a dimer, whereas GDH-BTN-His and GDH-BTC-His are monomeric. These results suggest that the intact N- and C-termini are required for GDH-BT to maintain thermostability and to form its dimer structure. The homology model of the GDH-BTWT single subunit was constructed based on the crystal structure of Bacillus megaterium GDH (PDB ID 3AY6), showing that GDH-BTWT has a Rossmann fold structure with its N- and C-termini located on the subunit surface, which suggests that His-tagging affected the native dimer structure. GDH-BTWT and GDH-BTN-His regenerated NADPH in a yeast reductase-mediated chiral synthesis reaction, suggesting that these enzymes can be used as catalysts in fine-chemical and pharmaceutical industries.

Preparation and Characterization of Cy5.5-conjugated Biocompatible Polymeric Micellar Nanoparticles for Optical Imaging (광학 영상을 위한 Cy5.5가 결합된 생체적합성 고분자 마이셀 나노입자의 제조 및 특성분석)

  • Kim, Hyo-Jeong;Kim, Byung-Jin;Lee, Ha-Yeong;Jung, Suk Hyun;Jeong, Seo-Young;Yuk, Soon-Hong;Shin, Byung-Cheo;Seong, Ha-Soo;Choi, Youn-Woong;Ha, Dae-Chul;Choi, Sun-Hang;Lee, Soo-Min
    • Journal of Pharmaceutical Investigation
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    • v.39 no.6
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    • pp.393-400
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    • 2009
  • PHEA (hydroxyethyl-aspartamide)-mPEG (methoy-polyethyleneglycol)-$C_{16}$ (hexadecylamine)-ED (ethylenediamine) was prepared as a drug delivery carrier. The structure and molecular weight of polymers were characterized by $^1H$-NMR and gel permeation chromatography. Micelle size and shape were measured by electro-photometer light scattering and transmission electron microscope. The mean diameter of micelles was 23 nm in aqueous solution. To evaluate the potential of these polymeric micelles as a drug carrier, PSI-mPEG-$C_{16}$-ED was conjugated with Cy5.5 for Near-Infrared Fluorescent (NIRF) based optical imaging. PSI-mPEG-$C_{16}$-ED-Cy5.5 was injected intravenously into mice (n=5) and in vivo NIRF imaging was performed during 48 h after injection. The biodistribution study at 24 h after injection showed the longcirculation property of PSI-mPEG-$C_{16}$-ED-Cy5.5. Therefore, PSI-mPEG-$C_{16}$-ED micelles could be a promising drug carrier and imaging agent.

Purification of Recombinant CTP-Conjugated Human prostatic acid phosphatase for activation of Dendritic Cell (수지상세포 활성화를 위한 세포투과 펩타이드가 결합된 재조합 전립성 산성 인산분해효소의 정제)

  • Yi, Ki-Wan;Ryu, Kang
    • KSBB Journal
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    • v.24 no.1
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    • pp.80-88
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    • 2009
  • Human prostatic acid phosphatase (PAP), with comprehensive homology to glandular kallikrein, are representative serum biomarkers of prostate cancer. Dendritic cell (DC), which is the potent antigen-presenting cells(APC) in the immune system, can induce strong T cell responses against viruses, microbial pathogens, and tumors. Therefore, the immunization using DC loaded with tumor-associated antigens is a powerful method for inducing anti-tumor immunity. The CTP (Cytoplasmic Transduction Peptide) technology developed by Creagene which can transport attached bio-polymers like nucleic acids or proteins into the cell with high permeation efficiency. As the active forms of PAP can mediate apoptotic processing, we used multimer forms of PAP as an inactive form for antigen pulsing of DCs. In this study, multimeric forms of CTP-rhPAP was obtained according to the advanced purification process and subsequently confirmed by gel filtration chromatography, western blot and Dynamic Light Scattering. Therefore, CTP-conjugated PA multimers transduced into the cytoplasm were efficiently presented on the cell surface without any harm effect on cells via MHC class I molecules and result in induction of a large number of effector cell.

Effects of Annealing Temperature on Thermal Properties of Glycidyl Azide Polyol-based Energetic Thermoplastic Polyurethane (글리시딜아자이드계 열가소성 폴리우레탄의 열적특성에 대한 열처리 조건의 영향)

  • Kim, Jeong Su;Kim, Du Ki;Kweon, Jeong Ohk;Lee, Jae Myung;Noh, Si Tae;Kim, Sun Young
    • Applied Chemistry for Engineering
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    • v.24 no.3
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    • pp.305-313
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    • 2013
  • In this study, we investigated effects of thermal annealing on the thermal properties and microphase separation behaviors of glycidyl azide-based thermoplastic polyurethane elastomers (ETPE). The GAP-based ETPEs were characterized by attenuated total reflectance-fourier transform infrared spectroscopy (ATR-FTIR), differential scanning calorimeter (DSC), dynamic mechanical analysis (DMA), and gel permeation chromatography (GPC). The effects of annealing temperature conditions ($80{\sim}130^{\circ}C$, 1 h or 24 h) on the properties of the ETPEs were investigated. The intensity of azide group absorption peak of ATR-FTIR spectra and the solubility of ETPE for methylene chloride and dimethylformamide solvent decreased after the annealing at $130^{\circ}C$ for 1 h and at $105^{\circ}C$ for 24 h. With increasing the annealing temperature from $80^{\circ}C$ to $110^{\circ}C$, the high temperature rubbery plateau region of storage modulus curves from DMA thermogram for GAP-based ETPEs was extended to the higher temperature.