• Title/Summary/Keyword: Dye affinity

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Isolation and Characterization of L-Ascorbic Acid-Producing Enzyme in Neurospora crassa (Neurospora crassa의 L-Ascorbic Acid 생산효소의 순수 분리 및 이의 특성에 관한 연구)

  • Kim, In-Sil;Lee, Yeon-Hee
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.132-138
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    • 1994
  • L-Ascorbic acid-producing enzyme in Neurospora crassa was found to exist in mitochondria and the activity of this enzyme was increased by the addition of D-fluconno-${\gamma}$-lactone or L-gulono-${\gamma}$-lactone in the media. L-Ascorbic acid-producin enzyme in N. crassa has been purified with ammonium sulfate precipitation. DEAE Sepharose CL-6B ion exchange chromatography. Sephacryl S-200 gel filtration chromatography and Reactive yellow 3-agarose dye affinity column chromatography. The specific activity of this enzyme was increased to 239.6 fold and the yield was 2.1%. The molecular weight of the native enzyme was 150.000 dalton when it was estimated with Sephacryl S-200 gel filtration chromatography. Its molecular weight appeared as 75.000 dalton by SDS-polyacrylamide gel electrophoresis. which suggested that this enzyme was consisted with two identical subunits. The optimal pH for this enzyme was 9.0 and the $K_m$ value for D-galactono-${\gamma}$-lactone was 0.073 mM.

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Color Characteristics of Silk Fabrics Dyed with Natural Red Dyes Without a Mordant (적색계 천연염료를 이용한 견직물 무매염 염색의 염색성)

  • Kim, Hyo-Jin;Lee, Joo-Hyeon
    • Journal of Digital Convergence
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    • v.16 no.12
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    • pp.593-602
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    • 2018
  • The purpose of this study was to investigate the coloring properties of non-mordant dyes by examining local literature on silk fabric dyeing using red-colored natural dyes. Natural dyes can be prepared from the following 8 materials: purple-fleshed sweet potato, Impatiens balsamina, mulberry, fruits of Ligustrum japonicum Thunb, guava leaves, dansam, hibiscus flowers, and pruned branches of Prunus persica. To examine the surface color calculated $L^*$, $a^*$, $b^*$, ${\Delta}E^*$, and K/S value and H V/C. The variables considered in the dyeing experiment were dye concentration, dyeing temperature, dyeing time, and number of dyeing iterations, which were varied to evaluate the dyeing properties and color characteristics. As a result, the abovementioned variables, dye affinity, and red color expression were directly proportional to one another. In this study, it was found that red coloring can be obtained with natural dyes; moreover, excellent dyeing was achieved without the need for repeated dyeing or mordanting processes, which cause environmental pollution.

Evaluation of Near-infrared Fluorescence-conjugated Peptides for Visualization of Human Epidermal Receptor 2-overexpressed Gastric Cancer

  • Jeong, Kyoungyun;Kong, Seong-Ho;Bae, Seong-Woo;Park, Cho Rong;Berlth, Felix;Shin, Jae Hwan;Lee, Yun-Sang;Youn, Hyewon;Koo, Eunhee;Suh, Yun-Suhk;Park, Do Joong;Lee, Hyuk-Joon;Yang, Han-Kwang
    • Journal of Gastric Cancer
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    • v.21 no.2
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    • pp.191-202
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    • 2021
  • Purpose: A near-infrared (NIR) fluorescence imaging is a promising tool for cancer-specific image guided surgery. Human epidermal receptor 2 (HER2) is one of the candidate markers for gastric cancer. In this study, we aimed to synthesize HER2-specific NIR fluorescence probes and evaluate their applicability in cancer-specific image-guided surgeries using an animal model. Materials and Methods: An NIR dye emitting light at 800 nm (IRDye800CW; Li-COR) was conjugated to trastuzumab and an HER2-specific affibody using a click mechanism. HER2 affinity was assessed using surface plasmon resonance. Gastric cancer cell lines (NCI-N87 and SNU-601) were subcutaneously implanted into female BALB/c nu (6-8 weeks old) mice. After intravenous injection of the probes, biodistribution and fluorescence signal intensity were measured using Lumina II (Perkin Elmer) and a laparoscopic NIR camera (InTheSmart). Results: Trastuzumab-IRDye800CW exhibited high affinity for HER2 (KD=2.093(3) pM). Fluorescence signals in the liver and spleen were the highest at 24 hours post injection, while the signal in HER2-positive tumor cells increased until 72 hours, as assessed using the Lumina II system. The signal corresponding to the tumor was visually identified and clearly differentiated from the liver after 72 hours using a laparoscopic NIR camera. Affibody-IRDye800CW also exhibited high affinity for HER2 (KD=4.71 nM); however, the signal was not identified in the tumor, probably owing to rapid renal clearance. Conclusions: Trastuzumab-IRDye800CW may be used as a potential NIR probe that can be injected 2-3 days before surgery to obtain high HER2-specific signal and contrast. Affibody-based NIR probes may require modifications to enhance mobilization to the tumor site.

Develop ECO-FREE high concentration Full black dye using transfer printing and application technology (전사날염용 ECO-FREE 고농도 Full Black 염료개발과 응용기술)

  • Cho, Ho-Hyun;Chung, Myung-Hee;Lee, A-Ram
    • Journal of the Korea Fashion and Costume Design Association
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    • v.19 no.2
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    • pp.39-48
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    • 2017
  • Transfer printing is a method to combine printing and dyeing technology by the use of sublimation. It is an environmentally-friendly printing method that saves costs, reduces the production processes by the omission of the washing process, and saves time by maintaining quality. Due to the development of transfer printing, a high value added printing technology is available now but color fastness to sublimation of the printing products is still low since there are few dyes that have an affinity to the fabrics and the application technology is still inadequate. Specially, in case of high concentration black dyes, eco-label type black dyes, which is a substitution for general dispersal dyes, have been developed while general dispersal black dyes are still used, creating issues such as color differences on the surface and back side of the fabrics and contamination by friction after transfer printing. There are also some restricted substances such as allergens. To address these issues, high concentration black dyes and application technology that are environmentally-friendly and that have over 16 K/S through the use of single dyes with excellent color fastness, fixation ability, and similar melting temperature were developed for this study.

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Cloning, Sequencing and Characterization of Acyltransferase Gene Involved in Exopolysaccharide Biosynthesis of Zoogloea ramigera 115SLR

  • Lee Sam-Pin;Troyano Esperanza;Lee Jin-Ho;Kim Hyun-Soo;Sinskey Anthony John
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1163-1168
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    • 2006
  • The recombinant plasmid pLEX2FP complements the mutation in Zoogloea ramigera 115MM1, and the complemented mutant produces an exopolysaccharide that shows higher affinity for the calcofluor dye than the exopolysaccharide from Z. ramigera 115SLR, resulting in higher fluorescence intensity under UV light. A compositional and structural analysis of the exopolysaccharide from Z. ramigera 115MM1 showed that the different fluorescent properties were due to a lower content of acetyl groups when compared with Z. ramigera 115SLR exopolysaccharide. These results were in agreement with a sequence analysis of the gene carried in the plasmid pLEX2FP, which appeared to encode an O-acyltransferase highly homologous to the 3-O-acyltransferase of Streptomyces mycarofaciens. The gene encoding the acyltransferase from Z. ramigera 115SLR was expressed as a GST-fusion protein with 70,000 daltons in E. coli.

Cell-Specific Targeting of Texas Red with Anti-Ep-CAM Antibody

  • Lee, Soo-Chul;Tae, Gun-Sik
    • Journal of Photoscience
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    • v.12 no.3
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    • pp.123-127
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    • 2005
  • The polyclonal antibody was generated against the peptide fragment of 62 amino acid residues (D 181-T242) near the COOH-terminal region of the extracellular domain of epithelial-cell adhesion molecule (Ep-CAM) and shown to be able to recognize Ep-CAM in competitive ELISA. Then, sulforhodamine 101 acid chloride (so called Texas red), a fluorescence dye, was conjugated to the affinity-purified anti-Ep-CAM antibody utilizing the reaction between the aliphatic amines of antibody and the sulfonyl chloride of Texas red. The molar ratio of Texas red to antibody was estimated to be approximately 1.86 by measuring optical densities at 280 nm and 596 nm, implying that the two molecules of Texas red at most were conjugated to antibody. The anti-Ep-CAM antibody-Texas red conjugate was then used for immunohistochemistry of CT-26 murine colon carcinoma cells. Based upon the fluorescence microscope images, anti-Ep-CAM antibody is able to deliver Texas red specifically to the surface of CT-26 cells on which Ep-CAM was actively expressed. This result indicates that anti-Ep-CAM antibody could be useful for the tissue-specific delivery of photosensitizers via antigen-antibody interaction.

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Adsorption of Non-degradable Eosin Y by Activated Carbon (활성탄에 의한 난분해성 염료인 Eosin Y의 흡착)

  • Lee, Min-Gyu;Kam, Sang-Kyu;Suh, Keun-Hak
    • Journal of Environmental Science International
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    • v.21 no.5
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    • pp.623-631
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    • 2012
  • The adsorption behavior of Eosin Y on activated carbon (AC) in batch system was investigated. The adsorption isotherm could be well fitted by the Langmuir adsorption equation. The kinetics of adsorption followed the pseudo-second-order model. The temperature variation was used to evaluate the values of free energy (${\Delta}G^{\circ}$), enthalpy (${\Delta}H^{\circ}$) and entropy (${\Delta}S^{\circ}$). The positive value of enthalpy change ${\Delta}H^{\circ}$ for the process confirms the endothermic nature of the process and more favourable at higher temperature, the positive entropy of adsorption ${\Delta}S^{\circ}$ reflects the affinity of the AC material toward Eosin Y and the negative free energy values ${\Delta}G^{\circ}$ indicate that the adsorption process is spontaneous. With the increase of the amount of AC, removal efficiency of Eosin Y was increased, but adsorption capacity was decreased. And adsorption capacity was increased with the decrease of particle size. With the increase of the amount of AC, removal efficiency of Eosin Y was increased, but adsorption capacity was decreased. And adsorption capacity was increased with the decrease of particle size.

Genomic Detection using Electrochemical Method (전기화학적 방법에 의한 유전자의 검출)

  • Choi, Yong-Sung;Lee, Kyung-Sup;Park, Dae-Hee
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.18 no.6
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    • pp.560-570
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    • 2005
  • In this paper, a microelectrode away DNA chip was fabricated on glass slide using photolithography technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5' end were immobilized on the gold electrodes by DNA arrayer utilizing the affinity between gold and sulfu. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Cyclic voltammetry in 5mA ferricyanide/ferrocyanide solution at 100 mV/s confirmed the immobilization of probe DNA on the gold electrodes. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic system.

The Purification and Characterization of a Bacillus stearothermophilus Methionine Aminopeptidase (MetAP)

  • Chung, Jae-Min;Chung, Il-Yup;Lee, Young-Seek
    • BMB Reports
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    • v.35 no.2
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    • pp.228-235
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    • 2002
  • Methionine aminopeptidase (MetAP) catalyzes the removal of an amino-terminal methionine from a newly synthesized polypeptide. The enzyme was purified to homogeneity from Bacillus stearothermophilus (KCTC 1752) by a procedure that involves heat precipitation and four sequential chromatographs (including DEAE-Sepharose ion exchange, hydroxylapatite, Ultrogel AcA 54 gel filtration, and Reactive red 120 dye affinity chromatography). The apparent molecular masses of the enzyme were 81,300 Da and 41,000 Da, as determined by gel filtration chromatography and sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), respectively. This indicates that the enzyme is comprised of two identical subunits. The MetAP specifically hydrolyzed the N-terminal residue of Met-Ala-Ser that was used as a substrate, and exhibited a strong preference for Met-Ala-Ser over Leu-Gly-Gly, Leu-Ser-Phe, and Leu-Leu-Tyr. The enzyme has an optimal pH at 8.0, an optimal temperature at $80^{\circ}C$, and pI at 4.1. The enzyme was heat-stable, as its activity remained unaltered when incubated at $80^{\circ}C$ for 45 min. The Km and Vmax values of the enzyme were 3.0mM and 1.7 mmol/min/mg, respectively. The B. stearothernmophilus MetAP was completely inactivated by EDTA and required $Co^{2+}$ ion(s) for activation, suggesting the metal dependence of this enzyme.

Effect of Sodium Bytyrate on Glycosylation of Recombinant Erythropoietin

  • Chung, Bo-Sup;Jeong, Yeon-Tae;Chang, Kern-hee;Kim, Jong-Soo
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1087-1092
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    • 2001
  • The effect of Sodium Butyrate (NaBu) on the N-linked oligosaccharide structure of Erythropoietin (EPO) was investigated. Recombinant human EPO was produced by CHO cells grown in an $MEM{\alpha}$ medium with or without 5 mM NaBu, and purified from the culture supernatants using a heparin-sepharose affinity column and immunoaffinity column. The N-linked oligosaccharides were released enzymatically and isolated by paper chromatography. The isolated oligosaccharides were then labeled with a fluorescent dye, 2-aminobenzamide, and analyzed with MonoQ anion exchange chromatography and GlycosepN amide chromatography for the assignment of a GU (glucose unit) vague. A glycan analysis by HPLC showed that the most significant characteristic effect of NaBu was a reduction in the proportion of glycans with Sri-and tetrasialylated oligodaccharides from $21.30\%$ (tri-) and $14.86\%$ (tetra-) in the control cultures (without NaBu) to $8.72\%$ (tri-) and $1.25\%$ (tetra-) in the NaBu-treated cultures, respectively. It was also found that the proportion of asialo-glycan increased from $12.54\%\;to\;23.6\%$ when treated with NaBu.

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