• 제목/요약/키워드: Enhanced fluorescence substrate

검색결과 8건 처리시간 0.044초

롤투롤 나노 복제 공정을 이용한 이차원 광결정 소자의 제작 (Fabrication of Two-dimensional Photonic Crystal by Roll-to-Roll Nanoreplication)

  • 김영규;변의현;장호영;김석민
    • 한국기계가공학회지
    • /
    • 제12권5호
    • /
    • pp.16-22
    • /
    • 2013
  • A two-dimensional photonic crystal structure was investigated using a roll-to-roll nanoreplication and physical vapor deposition processes for the inexpensive enhanced fluorescence substrate which is not sensitive to the polarization directions of excitation light source. An 8 inch silicon master having nano dot array with a diameter of 200 nm, a height of 100 nm and a pitch of 400 nm was prepared by KrF laser scanning lithography and reactive ion etching processes. A flexible polymer mold was fabricated by flat type UV replication process and a deposition of 10 nm nickel layer as an anti-adhesion layer. A roll mold was prepared by warping the flexible polymer mold on an aluminum roll base and a roll-to-roll UV replication process was carried out using the roll mold. After the deposition of ~ 100 nm $TiO_2$ layer on the replicated nano dot array, a 2 dimensional photonic crystal structure was realized with a resonance wavelength of 635 nm for both p- and s-polarized light sources.

Substrate Construes the Copper and Nickel Ions Impacts on the Mushroom Tyrosinase Activities

  • Gheibi, N.;Saboury, A.A.;Haghbeen, K.
    • Bulletin of the Korean Chemical Society
    • /
    • 제27권5호
    • /
    • pp.642-648
    • /
    • 2006
  • Mushroom tyrosinase (MT) structural changes in the presence of $Cu ^{2+}$ and $Ni ^{2+}$ were studied separately. Far-UV CD spectra of the incubated MT with the either of the metal ions indicated reduction of the well-ordered secondary structure of the enzyme. Increasing in the maximum fluorescence emission of anilinonaphthalene-8-sulfonic acid (ANS) was also revealing partial unfolding caused by the conformational changes in the tertiary structure of MT. Thermodynamic studies on the chemical denaturation of MT by dodecyl trimethylammonium bromide (DTAB) showed decrease in the stability of MT in the presence of $Cu ^{2+}$ or $Ni ^{2+}$ using their activation concentrations. Both activities of MT were also assessed in the presence of different concentrations of these ions, separately, with various monophenols and their corresponding diphenols. Kinetic studies revealed that cresolase activity on p-coumaric acid was boosted in the presence of either of the metal ions, but inhibited when phenol, L-tyrosine, or 4-[(4-methylphenyl)azo]-phenol was substrate. Similarly, catecholase activity on caffeic acid was enhanced in the presence of $Cu ^{2+}$ or $Ni ^{2+}$, but inhibited when catechol, L-DOPA, or 4-[(4-methylbenzo)azo]-1,2-benzenediol was substrate. Results of this study suggest that both cations make MT more fragile and less active. However, the effect of the substrate structure on the MT allosteric behavior can not be ignored.

Characterization of the active site and coenzyme binding pocket of the monomeric UDP- galactose 4'- epimerase of Aeromonas hydrophila

  • Agarwal, Shivani;Mishra, Neeraj;Agarwal, Shivangi;Dixit, Aparna
    • BMB Reports
    • /
    • 제43권6호
    • /
    • pp.419-426
    • /
    • 2010
  • Aeromonas hydrophila is a bacterial pathogen that infects a large number of eukaryotes, including humans. The UDP-galactose 4'-epimerase (GalE) catalyzes interconversion of UDP-galactose to UDP-glucose and plays a key role in lipopolysaccharide biosynthesis. This makes it an important virulence determinant, and therefore a potential drug target. Our earlier studies revealed that unlike other GalEs, GalE of A. hydrophila exists as a monomer. This uniqueness necessitated elucidation of its structure and active site. Chemical modification of the 6xHis-rGalE demonstrated the role of histidine residue in catalysis and that it did not constitute the substrate binding pocket. Loss of the 6xHis-rGalE activity and coenzyme fluorescence with thiol modifying reagents established the role of two distinct vicinal thiols in catalysis. Chemical modification studies revealed arginine to be essential for catalysis. Site-directed mutagenesis indicated Tyr149 and Lys153 to be involved in catalysis. Use of glycerol as a cosolvent enhanced the GalE thermostability significantly.

Perfluoropolyether (PFPE)로 처리된 표면의 생물오손 방지 특성 연구 (Study on Anti-biofouling Properties of the Surfaces Treated with Perfluoropolyether (PFPE))

  • 박수인;권순일;이영민;고원건;하종욱;이상엽
    • 공업화학
    • /
    • 제23권1호
    • /
    • pp.71-76
    • /
    • 2012
  • 해조류 및 따개비 등의 해양 생물에 의한 선박 및 해양 구조물 표면의 생물오손(biofouling)은 선박 운영비를 증가시키고 구조물을 유지, 보수하는데 어려움을 가져왔다. 본 논문에서는 이러한 생물오손 방지 또는 생물오손 제거(fouling-release)를 향상시키는 방안으로 불소계 화합물인 perfluoropolyether (PFPE)를 이용하여 해양 생물의 표면 점착을 억제하는 방법이 연구되었다. 우선 생물오손을 예측할 수 있는 지표로서 물방울 접촉각이 측정되었다. 아민그룹으로 처리 된 친수성 표면이 갖는 $46.7^{\circ}$의 물방울 접촉각이 PFPE 처리 후 $64.5^{\circ}$로 상승하여 표면의 혐수성이 증가하였다. 이로 인해 초기에 따개비 포자 및 해양 미생물의 점착이 친수성 카르복실 표면과 비교시 약 15% 억제되었다. 또한 표면 코팅시 평탄면이 형성되어 PDMS로 처리된 표면 굴곡이 있는 표면보다 점착된 미생물의 제거가 용이하였다. 이러한 점착 억제 특성은 물리화학적 방법을 통해 측정된 물성들과 비교, 분석되었으며, 표면에 점착된 미생물의 염색을 통한 형광도 측정을 통해 표면 점착도가 정량적으로 분석되었다. PFPE가 갖는 가공의 용이성과 저독성 특성으로 인해 PFPE는 향후 단기간 생물학적 방오염이 필요한 해양 구조물 이외에 단백질 점착 억제가 요구되는 의료용 장비 등에도 활용될 수 있을 것으로 기대된다.

Microcontact Printing of Biotin for Selective Immobilization of Streptavidin-fused Proteins and SPR Analysis

  • Lee, Sang-Yup;Park, Jong-Pil;Lee, Seok-Jae;Park, Tae-Jung;Lee, Kyung-Bok;Park, Insung S.;Kim, Min-Gon;Chung, Bong-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • 제9권2호
    • /
    • pp.137-142
    • /
    • 2004
  • In this study, a simple procedure is described for patterning biotin on a glass substrate and then selectively immobilizing proteins of interest onto the biotin-patterned surface. Microcontact printing (CP) was used to generate the micropattern of biotin and to demonstrate the selective immobilization of proteins by using enhanced green fluorescent protein (EGFP) as a model protein, of which the C-terminus was fused to a core streptavidin (cSA) gene of Streptomyces avidinii. Confocal fluorescence microscopy was used to visualize the pattern of the immobilized protein (EGFP-cSA), and surface plasmon resonance was used to characterize biological activity of the immobilized EGFP-cSA. The results suggest that this strategy, which consists of a combination of $\mu$CP and cSA-fused proteins. is an effective way for fabricating biologically active substrates that are suitable for a wide variety of applications. one such being the use in protein-protein assays.

Biochemical Reactions on a Microfluidic Chip Based on a Precise Fluidic Handling Method at the Nanoliter Scale

  • Lee, Chang-Soo;Lee, Sang-Ho;Kim, Yun-Gon;Choi, Chang-Hyoung;Kim, Yong-Kweon;Kim, Byung-Gee
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • 제11권2호
    • /
    • pp.146-153
    • /
    • 2006
  • A passive microfluidic delivery system using hydrophobic valving and pneumatic control was devised for microfluidic handling on a chip. The microfluidic metering, cutting, transport, and merging of two liquids on the chip were correctly performed. The error range of the accuracy of microfluid metering was below 4% on a 20 nL scale, which showed that microfluid was easily manipulated with the desired volume on a chip. For a study of the feasibility of biochemical reactions on the chip, a single enzymatic reaction, such as ${\beta}-galactosidase$ reaction, was performed. The detection limit of the substrate, i.e. fluorescein $di-{\beta}-galactopyranoside$ (FDG) of the ${\beta}-galactosidase$ (6.7 fM), was about 76 pM. Additionally, multiple biochemical reactions such as in vitro protein synthesis of enhanced green fluorescence protein (EGFP) were successfully demonstrated at the nanoliter scale, which suggests that our microfluidic chip can be applied not only to miniaturization of various biochemical reactions, but also to development of the microfluidic biochemical reaction system requiring a precise nano-scale control.