• 제목/요약/키워드: l-ethyl-3-(3-dimethylaminopropyl) carbodiimide

검색결과 7건 처리시간 0.019초

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

  • Lee, Kyoung
    • Journal of Microbiology and Biotechnology
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    • 제8권4호
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    • pp.416-421
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    • 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.

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

  • 조승희;연경호;김가영;심준목;문승현
    • 대한환경공학회지
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    • 제27권12호
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    • pp.1263-1269
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    • 2005
  • 용액 상에서 페놀류에 대해 95%의 분해특성을 지니는 동식물세포유래의 효소(horseradish peroxidase, HRP, EC 1.11.1.7)를 다공성 탄소 전극에 고정화시키고 이를 전기화학 반응기에 도입하여 전극반응에 의해 연속적으로 발생되는 과산화수소를 이용하여 페놀의 분해를 수행하였다. FT-IR 분석을 통해 다공성탄소전극 표면에 HRP의 아민기와 펩티드 결합을 위한 카르복실기가 생성되었음을 확인하였고 가교제(coupling agent)로 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride(EDC)를 이용하여 공유결합으로 고정화시켰다. 또한 HRP를 활성화 시켜 페놀을 처리하기 위해 전해질로 사용된 인산염 완충용액의 농도($10{\sim}200$ mM)와 pH($5.0{\sim}8.0$), 외부 산소 주입량($10{\sim}50$ mL/min)및 potentiostat/galvanosta에 의한 외부 공급전압($-0.2{\sim}-0.8$ volt, vs. Ag/AgCl)의 조건을 달리하며 RVC 전극 표면에서 발생되는 과산화수소 농도 및 전류효율을 고려하여 최적 자체 발생조건을 결정하였다. HRP가 고정화된 RVC 전극은 초기 고정화된 HRP 활성에 대해 4주 동안 89%의 상대적 효소 활성도(relatively enzymatic activity)를 지니는 안정한 전극임을 확인하였으며 실험실 스케일의 연속식 전기화학 반응기에 도입되어 최적 과산화수소의 발생조건에서 86%의 분해 효율을 보였다.

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
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    • 제8권2호
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    • pp.112-116
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    • 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
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    • 제8권4호
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    • pp.227-232
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    • 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
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    • 제11권5호
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    • pp.368-374
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    • 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
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    • 제13권3호
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    • pp.167-175
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    • 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.

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

  • 김민석;박참진;김수환;임홍국;김용진
    • Journal of Chest Surgery
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    • 제43권6호
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    • pp.576-587
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    • 2010
  • 배경: 이종조직판막이나 조직이 임상적으로 이용되기 위해서는 조직의 장기간의 내구성이 동반된 효과적인 보존과 적절한 고정과정이 필요하다. 본 연구에서는 소심낭을 sodium dodecyl sulfate (SDS)와 N-lauroylsarcosinate를 이용하여 무세포화하고, 알파-갈락토시다아제를 처리한 뒤, 이를 다양한 방법으로 고정함으로써 조직손상과 면역반응을 최소화할 수 있는 효과적인 우심낭 보존 방법에 대해 알아보고자 하였다. 대상 및 방법: 우심낭으로부터 이종 이식 보철편을 채취한 뒤 SDS와 N-lauroylsarcosinate를 이용하여 무세포화 하였고, 알파-갈락토시다아제를 처리하였다. 이 두 가지 군에 대하여 각기 다른 조건 즉 glutaraldehyde (GA), l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC)/Nhydroxysuccinamide (NHS)의 처리를 달리하여 고정하였다. 이후 각 군들의 물리적 성상, 인장강도, 열 안정성 검사, 세포독성 검사, 프로나아제 저항 시험과 Pronase-Ninhydrin 시험, 투과도와 유순도 검사, purpald 검사 등을 시행하였으며, H&E 염색과 DNA 정량, 알파-갈 항원결정인자 염색 등도 함께 시행하였다. 결과: GA 단독 고정군은 EDC/NHS 고정군에 비해 물리적인 성상과 열안정성에서 더 우수하였고, EDC/NHS와 GA의 이중 고정군은 EDC/NHS 고정군에 비해 물리적인 성상과 열안정성에서 더 우수하였으며, GA단독 고정군과 비교해서 열안정성에서 더 우수하였다. 프로나아제 저항 시험과 Pronase-Ninhydrin 시험을 통해 알아본 crosslinking 정도는, GA 고정 군과 EDC/NHS의 이중 고정군이 EDC/NHS 고정군에 비해 높은 것으로 관찰되었다. EDC/NHS와 GA의 이중 고정군은 GA 고정군에 비해 투과도와 유순도가 다소 증가하였으나, GA 고정군에 비해 인장강도가 높고 세포 독성이 감소하는 것으로 관찰되었다. 결론: GA와 EDC/NHS의 이중고정을 통해서, GA단독 고정의 독성을 줄이면서 효과적인 crosslinking을 시행할 수 있음을 확인하였다. 향후 생체내 시험을 통해서 GA와 EDC/NHS의 이중고정이 석회화의 감소와 조직부전에 어떠한 영향을 미치는지 추가적인 연구와 검증이 필요하다.