• Title/Summary/Keyword: Ion exchange chromatography(IEC)

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Determination of Adsorption Isotherms and Separation of L-arabinose and D-ribose in Cation Exchange Chromatography and HPLC (양이온 교환 크로마토그래피와 HPLC에서의 L-arabinose와 D-ribose의 분리 및 등온 흡착곡선 결정)

  • Jeon, Young-Ju;Kim, In-Ho
    • KSBB Journal
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    • v.23 no.1
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    • pp.31-36
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    • 2008
  • The use of L-carbohydrates and their corresponding nucleosides in medicinal application has greatly increased. For example L-ribose has been much in demand as the starting material for curing hepatitis B. High performance liquid chromatography (HPLC) method was studied for the analysis of ribose and arabinose fractions from ion exchange chromatography (IEC). Dowex Monosphere 99 Ca/320 resin was packed in IEC to separate ribose and arabinose under various operating conditions. $NH_{2}$ and sugar HPLC columns were then used to analyze the fractions from the IEC column. Pulse input method (PIM) was also used to measure adsorption isotherms of ribose and arabinose in the Dowex column and HPLC columns. Experimental results and simulations by ASPEN chromatography were compared with fair agreement.

Effects of Extraction Method on the Histidine Containing Low Molecular Weight Peptide and Pro-oxidants Contents of Tuna Boiled Extracts (참치자숙액 추출물 중의 히스티딘계 저분자 펩타이드 및 산화촉진물질 함량에 미치는 추출방법의 영향)

  • Kang, Ok-Ju
    • Korean journal of food and cookery science
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    • v.24 no.3
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    • pp.349-357
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    • 2008
  • In an effort to augment extractability of carnosine and anserine at the levels of pro-oxidants such as iron and protein in Tuna boiled extracts(Skipjack, Yellowfin and Bigeye), we assessed the effects of heated and ion exchange chromatography(IEC) and ultrafiltration(UF) using a MW 500 cut-off(500 MWCO). We also evaluated the antioxidant activity of these extracts processed as free radical scavengers and reducing agents. Tuna boiled extracts of dark and ordinary muscle protein and total iron were reduced, whereas carnosine and anserine concentrations and antioxidant activity were increased. The carnosine and anserine concentrations of the ion exchange and permeate UF(IEC-UF) extracts were higher than those observed in the heated and permeate UF(heat-UF), whereas the protein and total iron contents were lower than that observed in the heat-UF. The quantity of carnosine and anserine in ordinary muscle was higher than that detected in dark muscle. HPLC analysis and SDS-PAGE were shown to removes the effect of UF on high molecular weight impurities in the tuna boiled extracts. The major free amino acids(FFAs) from Skipjack, Yellowfin and Bigeye tuna IEC-UF extracts were anserine, histidine and carnosine. These three peptides constituted more than 80~85%. of the detected amino acid. The IEC-UF treated ordinary muscle extracts evidenced the highest levels of DPPH radical scavenging activity and the highest levels of reducing power among the various extracts. The IEC-UF extracts evidenced a DPPH radical scavenging effect equal to that of 1mM ascorbic acid.

Effects of Extraction Methods on Histidine-containing Low-molecular Weight Peptides and Pro-oxidants Contents in Tuna Thunnus Extracts (다랑어(Thunnus) 추출물 중의 Histidine 함유 저분자 펩타이드 및 산화촉진물질 함량에 미치는 추출방법의 영향)

  • Kim, Hong-Kil;Song, Ho-Su
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.6
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    • pp.684-693
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    • 2017
  • We investigated methods for extracting histidine-containing low-molecular-weight (LMW) peptides such as anserine, carnosine and histidine from the edible meat of tuna byproducts. Extracts were treated by several methods including heat treatment ($80^{\circ}C$, 10 min), DOWEX ion exchange (IEC), ultrafiltration (UF), and carboxymethyl (CM)-cellulose column chromatography (IEC+CMC); then the levels of protein, total iron, histidine, carnosine, and anserine were measured. Extracts treated with IEC+CMC using CM-cellulose were analyzed for total iron, protein, histidine, and anserine content, which were $6.27{\pm}0.26mg/mL$, $5.20{\pm}0.21{\mu}g/mL$, 0.80 mg/mL, 0.208 mg/mL, and 4.40 mg/mL, respectively, in yellowfin tuna; and $9.05{\pm}0.82mg/mL$, $4.06{\pm}0.20{\mu}g/mL$, 1.62 mg/mL, 0.012 mg/mL, and 7.28 mg/mL in bigeye tuna. By comparison in IEC-UF treated extracts, protein, total iron, and histidine content decreased by 43%, 73%, and 27% in yellowfin and 0.4%, 54%, and 23% in bigeye tuna, wheres carnosine and anserine content increased by 22% and 17%, respectively. Freeze-dried (FD) extracts exhibited similar trends as non-dried extracts, i.e., dipeptide content increased with purification steps, whereas pro-oxidant (total iron and protein) content decreased. IEC+CMC treated FD extracts had the highest anserine, content, and the greatest reductuion in pro-oxidants.

Purification of fusion ferritin using silica powder and DEAE chromatography

  • Heo, Yun-Seok;Kim, Seong-Gyu;Jeong, Eun-Mi;Kim, In-Ho
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.510-513
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    • 2002
  • Iron is an essential nutrient for most organisms, which supplied to them in a protein-iron complex known as ferritin. Ferritins are multimeric proteins those are consisted of spherical shell of 24 subunits defining a cavity of about 8nm in diameter. Soluble form of ferritin was separated from disrupted cells, followed by silica powder adsorption. Ferritin was recovered from silica-poweder by distiiled water, which was applied to DEAE anion exchage chromatography. Collected fractions from the DEAE column were assayed to gain the amount and the purity of ferritin by using GF-HPLC.

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Fractionation and Enzymatic Characterization of Endoprotease and Exopeptidase from Crude Extracts of Cuttlefish Sepia officinalis Hepatopancreas

  • Kim, Min Ji;Kim, Hyeon Jeong;Kim, Ki Hyun;Heu, Min Soo;Lee, Jung Suck;Kim, Jin-Soo
    • Fisheries and Aquatic Sciences
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    • v.15 no.4
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    • pp.283-291
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    • 2012
  • This study examines the optimal fractionation method and conditions for the isolation of endoprotease- and exopeptidase-active fractions from crude extracts of cuttlefish hepatopancreas (HP) using four fractionation methods: ammonium sulfate fractionation (ASF), polyethylene glycol fractionation (PGF), ion exchange chromatography (IEC), and gel filtration chromatography (GFC). Total endoprotease activity highest in the fraction II (concentrate of fractions 34-42; 842.60 U) of GFC, followed by fraction III (40-60% ammonium sulfate fraction; 670.25 U) of ASF, fraction I (concentrate of fractions 8-12; 436.89 U) of IEC, and fraction II (10-20% polyethylene glycol; 307.31 U) of PGF. Total exopeptidase activity of these fractions was highest in fraction II (2,704.70 U) of GFC, fraction III (2,110.50 U) of ASF, fraction III (1,605.60 U) of PGF, and fraction II (concentrate of fractions 38-44; 1,196.22 U) of IEC. These results showed that fraction II of GFC had the highest activity toward both exopeptidase and endoprotease, with exopeptidase activity being 3.21 times higher than of endoprotease. These results suggest cuttlefish HP could be used as a potential source for the extraction of exopeptidase, an enzyme capable of catalyzing the cleavage of N- and C-terminal amino acids in polypeptides, Like endoprotease, the most efficient method for separating exopeptide-active fractions was GFC.

Synthesis and Characterization of Di and Triblock Copolymers Containing a Naphthalene Unit for Polymer Electrolyte Membranes (고분자전해질 막을 위한 나프탈렌 단위를 포함하는 디 및 트리 블록공중합체의 합성 및 특성분석)

  • KIM, AERHAN
    • Transactions of the Korean hydrogen and new energy society
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    • v.27 no.6
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    • pp.660-669
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    • 2016
  • A fluorinated-sulfonated, hydrophobic-hydrophilic copolymer was planed subsequently synthesized using typical nucleophilic substitution polycondensation reaction. A novel AB and ABA (or BAB) block copolymers were synthesized using sBCPSBP (sulfonated 4,4'-bis[4-chlorophenyl)sulfonyl]-1,1'-biphenyl), DHN (1,5-dihydroxynaphthalene), DFBP (decafluorobiphenyl) and HFIP (4,4'-hexafluoroisopropylidenediphenol). All block copolymers were easily cast and made into clear films. The structure and synthesized copolymers and corresponding membranes were analyzed using GPC (gel permeation chromatography), $^1H$-NMR ($^1H$ nuclear magnetic resonance) and FT-IR (Fourier transform infrared). TGA (Thermogravimetric analysis) and DSC (differential scanning calorimetry) analysis showed that the prepared membranes were thermally stable, so that elevated temperature fuel cell operation would be possible. Hydrophobic/hydrophilic phase separation and clear ionic aggregate block morpology was confirmed in both triblock and diblock copolymer in AFM (atomic force microscopy), which may be highly related to their proton transport ability. A sulfonated BAB triblock copolymer membrane with an ion-exchange capacity (IEC) of 0.6 meq/g has a maximum ion conductivity of 40.3 mS/cm at $90^{\circ}C$ and 100% relative humidity.

The Effect of Carnosine Extracted from Eels Anguilla japonica on Oxidative DNA Damage Induced by Hydrogen Peroxide and the DNA Repair Capacity of Human Leukocytes (뱀장어(Anguilla japonica) 추출 Carnosine이 과산화수소로 유도된 인체 백혈구의 DNA 손상과 Repair에 미치는 효과)

  • Song, Ho-Su
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.5
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    • pp.520-526
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    • 2017
  • Carnosine was recently reported to protect against the DNA damage induced by oxidative stress. In this study, we investigated the protective effect of eel Anguilla japonica carnosine extracts prepared using different methods (heat treatment extracts, HTEs; ion exchange chromatography, IEC; ultrafiltration permeation, UFP) on leukocyte DNA damage using the comet assay. Human leukocytes were incubated with extracts of eel carnosine at concentrations (of 10, 50, $100{\mu}g/mL$), and then subjected to an oxidative stimulus [$200{\mu}M$ hydrogen peroxide ($H_2O_2$)]. Pretreatment of the cells for 30 min with carnosine significantly reduced the genotoxicity of $H_2O_2$ measured as DNA strand breaks. The protective effects of the three types of extract (HTE, IEC, and UFP) increased with concentration. At the highest concentration (100 g/mL). there were no statistical differences in oxidative damage between each extract treatment and PBS-treated negative controls. When leukocytes were incubated with carnosine for 30 min after exposure to $H_2O_2$. the protective ability of each extract changed. Therefore, eel carnosine inhibits the $H_2O_2$ induced damage to cellular DNA in human leukocytes, supporting the protective effect of this compound against oxidative damage.

Antioxidant Effect of Anserine Extracted from Salmon (Oncorhynchus keta) (연어(Oncorhynchus keta) 추출 anserine의 항산화 효과)

  • Min, Hye-Ok;Song, Ho-Su
    • Journal of Food Hygiene and Safety
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    • v.34 no.4
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    • pp.396-403
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    • 2019
  • Ion-exchange chromatography and ultrafiltration were used to extract anserine from salmon (Oncorhynchus keta). The salmon anserine showed DPPH radical scavenging activity in the range of 7.30% to 31.05% in a dose-dependent manner. This reducing power of salmon anserine also increased as the concentration increased. Metal chelate activity, superoxide dismutase - like activity, and thiobarbituric acid reactive substances assay showed similar results. The anserine also suppressed the increment of the peroxide value and linoleic acid during storage periods. These results suggest that salmon anserine might be useful as a natural antioxidant in various foodstuffs.

Purification and Characterization of Antibacterial Compound Produced by Bacillus subtilis MJP1 (Bacillus subtilis MJP1이 생산하는 항세균 물질의 분리.정제 및 특성규명)

  • Yim, Eun-Jung;Yang, Eun-Ju;Chang, Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.38 no.1
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    • pp.84-92
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    • 2010
  • Antibacterial compound from Bacillus subtilis MJP1 was purified using C18 Sep-Pak cartridge, ion exchange chromatography, and gel filtration chromatography. The purified antibacterial compound showed antibacterial activity against Listeria monocytogenes, Bacillus subtilis, Staphylococcus aureus subsp. aureus, and Enterococcus faecalis. The purified antibacterial compound was found to be stable at $100^{\circ}C$ for 5 min and in the pH range of 3.0~9.0, but it was unstable at pH 10.0. It was inactivated by proteinase K and pronase E, and heat treatment at $121^{\circ}C$ for 15 min, but it was stable with lipase and $\alpha$-amylase treatment, which indicated its proteineous nature. Ultra performance liquid chromatography and electrospray ionization tandem mass spectrometry analysis were used to identify the purified antibacterial compound and confirmed the existence of two peptides (3356.54 Da, 3400.5244 Da).