• Title/Summary/Keyword: l-ethyl-3-(3-dimethylaminopropyl) carbodiimide

Search Result 7, Processing Time 0.018 seconds

Involvement of Electrostatic Interactions between the Components of Toluene Dioxygenase from Pseudomonas putida Fl

  • Lee, Kyoung
    • Journal of Microbiology and Biotechnology
    • /
    • v.8 no.4
    • /
    • pp.416-421
    • /
    • 1998
  • A carboxyl group modifier, l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) was used to study the interactions between three components of toluene dioxygenase (TDO) from Pseudomonas putida FI. $Ferredoxin_{TOL}$ activity was increased by the treatment with EDC; however, the activity was rapidly declined in the prolonged incubation. In covalent cross-linking experiments with EDC, $Ferredoxin_{TOL}$ made a one-to-one complex with $Reductase_{TOL}$ or the large subunit of $ISP_{TOL}$. These results provide evidence for the involvement of electrostatic interactions in the TDO electron transfer system.

  • PDF

Electrochemical Degradation of Phenol by Using Reticulated Vitreous Carbon Immobilized Horseradish Peroxidase (Horseradish Peroxidase가 고정화된 다공성 탄소 전극을 이용한 페놀의 전기화학적 분해)

  • Cho, Seung-Hee;Yeon, Kyeong-Ho;Kim, Gha-Young;Shim, Joon-Mok;Moon, Seung-Hyeon
    • Journal of Korean Society of Environmental Engineers
    • /
    • v.27 no.12
    • /
    • pp.1263-1269
    • /
    • 2005
  • Horseradish peroxidase, had the phenol degradation rate of 95% in aqueous phase, was covalently immobilized on the surface of reticulated vitreous carbon(RVC) and the degradation of phenol was performed with in situ generated $H_2O_2$-immobilized HRP complex in an electrochemical reactor. The incorporation of carboxylic group on the RVC surface was confirmed by FT/IR spectrometry and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride(EDC) was used for peptide bonds between the carboxylic groups on the RVC surface and amine groups from HRP. The optimal conditions of in situ $H_2O_2$ generation such as concentration($10{\sim}200$ mM) and pH($5.0{\sim}8.0$) of electrolyte, supply of $O_2(10{\sim}50$ mL/min) and applied voltage($-0.2{\sim}-0.8$ volt, vs. Ag/AgCl) from potentiostat/galvanostat were determined by concentration of hydrogen peroxide and current efficiency. It was observed that the RVC immobilized HRP was stable maintaining 89% of the initial activity during 4 weeks. The phenol degradation rate of 86% was attained under the optimal condition of in situ $H_2O_2$ generation.

Surface Plasmon Resonance Immunosensor for Detection of Legionella pneumophila

  • Oh, Byung-Keun;Lee, Woochang;Bae, Young-Min;Lee, Won-Hong;Park, Jeong-Woo
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.8 no.2
    • /
    • pp.112-116
    • /
    • 2003
  • An immunosensor based on surface plasmon resonance (SPR) onto a protein G layer by Self-assembly technique was developed for detection of Legionella pneumophila. The protein G layer by self-assembly technique was fabricated on a gold (Au) surface by adsorbing the 11-mercaptoundecanoic acid (MUA) and an activation process for the chemical binding of the free amino (-NH$_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC) in series. The formation of the protein G layer by self-assembly technique on the Au Substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The Surface topographies of the fabricated thin films on an Au substrate were also analyzed by using an atomic force microscope (AFM). Consequently, an immunosensor for the detection of L. pneumophila using SPR was developed with a detection limit of up to 10$^2$CFU per mL.

Nano-scale Probe Fabrication Using Self-assembly Technique and Application to Detection of Escherichia coli O157:H7

  • Oh, Byung-Keun;Lee, Woochang;Lee, Won-Hong;Park, Jeong-Woo
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.8 no.4
    • /
    • pp.227-232
    • /
    • 2003
  • A self-assembled monolayer of protein G was fabricated to develop an immunosensor based on surface plasmon resonance (SPR), thereby improving the performance of the antibodybased biosensor through immobilizing the antibody molecules (lgG). As such, 11-mercaptoundecanoic acid (11-MUA) was adsorbed on a gold (Au) support, while the non-reactive hydrophilic surface was changed through substituting the carboxylic acid group (-COOH) in the 11-MUA molecule using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrocholide (EDAC). The formation of the self-assembled protein G layer on the Au substrate and binding of the antibody and antigen were investigated using SPR spectroscopy, while the surface topographies of the fabricated thin films were analyzed using atomic force microscopy (AFM). A fabricated monoclonal antibody (Mab) layer was applied for detecting E. coli O157:H7. As a result, a linear relationship was achieved between the pathogen concentration and the SPR angle shift, plus the detection limit was enhanced up to 10$^2$ CFU/mL.

Artificial Dermis Composed of Gelatin, Hyaluronic Acid and (1\longrightarrow3),(1\longrightarrow6)-$\beta$-Glucan

  • Lee, Sang-Bong;Jeon, Hyun-Wook;Lee, Young-Woo;Cho, Seong-Kwan;Lee, Young-Woo;Song, Kang-Won;Park, Moon-Hyang;Hong, Sung-Hwa
    • Macromolecular Research
    • /
    • v.11 no.5
    • /
    • pp.368-374
    • /
    • 2003
  • Porous scaffolds composed of gelatin and polysaccharides such as hyaluronic acid and $\beta$-glucan were prepared by using the freeze-drying method after cross-linking with l-ethyl-(3-3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). The scaffold had an inter-connected pore structure with the sufficient pore size for use as a support for the growth of fibroblasts. Results for the contact angle and cell attachment confirmed that high gelatin content in a mixture was suitable for cellular attachment and distribution in two- or three-dimensional fibroblast cultures. However, the addition of polysaccharides aroused the synergistic effects of morphologic and mechanical property of gelatin-based scaffolds. To prepare the artificial dermis for the wound dressing to mimic the normal human dermal skin, fibroblasts were isolated from a child's foreskin, and cultured in gelatin-based scaffolds. An in vivo study showed that the artificial dermis containing the fibroblasts enhanced the wound healing rate and re-epithelialization of a full-thickness skin defect rather than the acellular scaffold after one week.

Self-Assembled Nanoparticles of Bile Acid-Modified Glycol Chitosans and Their Applications for Cancer Therapy

  • Kim Kwangmeyung;Kim Jong-Ho;Kim Sungwon;Chung Hesson;Choi Kuiwon;Kwon Ick Chan;Park Jae Hyung;Kim Yoo-Shin;Park Rang-Won;Kim In-San;Jeong Seo Young
    • Macromolecular Research
    • /
    • v.13 no.3
    • /
    • pp.167-175
    • /
    • 2005
  • This review explores recent works involving the use of the self-assembled nanoparticles of bile acid-modified glycol chitosans (BGCs) as a new drug carrier for cancer therapy. BGC nanoparticles were produced by chemically grafting different bile acids through the use of l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC). The precise control of the size, structure, and hydrophobicity of the various BGC nanoparticles could be achieved by grafting different amounts of bile acids. The BGC nanoparticles so produced formed nanoparticles ranging in size from 210 to 850 nm in phosphate-buffered saline (PBS, pH=7.4), which exhibited substantially lower critical aggregation concentrations (0.038-0.260 mg/mL) than those of other low-molecular-weight surfactants, indicating that they possess high thermodynamic stability. The SOC nanoparticles could encapsulate small molecular peptides and hydrophobic anticancer drugs with a high loading efficiency and release them in a sustained manner. This review also highlights the biodistribution of the BGC nanoparticles, in order to demonstrate their accumulation in the tumor tissue, by utilizing the enhanced permeability and retention (EPR) effect. The different approaches used to optimize the delivery of drugs to treat cancer are also described in the last section.

Study on Effective Preservation of Bovine Pericardium Using Decellulariation and ${\alpha}$-galactosidase for Eliminating Xenoreactive Antigen (이종 항원 제거를 위한 무세포화와 알파-갈락토시다아제를 이용한 효과적인 우심낭 보존 방법에 관한 연구)

  • Kim, Min-Seok;Park, Cham-Jin;Kim, Soo-Hwan;Lim, Hong-Gook;Kim, Yong-Jin
    • Journal of Chest Surgery
    • /
    • v.43 no.6
    • /
    • pp.576-587
    • /
    • 2010
  • Background: Effective decellularization and fixation process is critical, in order to use xenogenic valves clinically. In the present study, we decellularized bovine pericardium using sodium dodecyl sulfate (SDS) and N-lauroyl sarcosinate, treated with $\alpha$-galactosidase, and then fixed in various manners, to find out the most effective tissue preservation & fixation procedure. Material and Method: Bovine pericardium was decellularized with SDS and N-lauroyl sarcosinate, and treated with $\alpha$-galactosidase. Both groups were fixed differently, by varying glutaraldehyde (GA) or EDC (1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide)/N-hydroxysuccinamide (NHS) treatment conditions. Thereafter, physical examination, tensile strength test, thermal stability test, cytotoxicity test, pronase test, pronase-ninhydrin test, purpald test, permeability test, compliance test, H&E staining, DNA quantification, and $\alpha$-galactose staining were carried out to each groups. Result: GA fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, EDC/NHS-GA dual fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, and showed better thermal stability than GA fixed groups. In pronase test and pronase-ninhydrin test, GA fixed groups and EDC/NHS-GA dual fixed groups showed stronger crosslinks than EDC/NHS groups. Permeability and compliance tended to increase in EDC/NHS-GA dual fixed groups, compared to GA fixed groups. But, EDC/NHS-GA dual fixed groups had stronger tensile strength and lower cytotoxicity than GA fixed groups. Conclusion: We have verified that EDC/NHS-GA dual fixation can make effective crosslinks and lower the toxicity of GA fixation. Henceforth, we will verify if EDC/NHS-GA dual fixation can lower calcifications & tissue failure in vivo experiment.