• 제목/요약/키워드: Cell surface display

검색결과 205건 처리시간 0.033초

Biodegradation of Organophosphate Pesticide Using Recombinant Cyanobacteria with Surface- and Intracellular-Expressed Organophosphorus Hydrolase

  • Chungjatupornchai, Wipa;Fa-Aroonsawat, Sirirat
    • Journal of Microbiology and Biotechnology
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    • 제18권5호
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    • pp.946-951
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    • 2008
  • The opd gene, encoding organophosphorus hydrolase (OPH) from Flavobacterium sp. capable of degrading a wide range of organophosphate pesticides, was surface- and intracellular-expressed in Synechococcus PCC7942, a prime example of photoautotrophic cyanobacteria. OPH was displayed on the cyanobacterial cell surface using the truncated ice nucleation protein as an anchoring motif. A minor fraction of OPH was displayed onto the outermost surface of cyanobacterial cells, as verified by immunostaining visualized under confocal laser scanning microscopy and OPH activity analysis; however, a substantial fraction of OPH was buried in the cell wall, as demonstrated by proteinase K and lysozyme treatments. The cyanobacterial outer membrane acts as a substrate (paraoxon) diffusion barrier affecting whole-cell biodegradation efficiency. After freeze-thaw treatment, permeabilized whole cells with intracellular-expressed OPH exhibited 14-fold higher bioconversion efficiency ($V_{max}/K_m$) than that of cells with surface-expressed OPH. As cyanobacteria have simple growth requirements and are inexpensive to maintain, expression of OPH in cyanobacteria may lead to the development of a low-cost and low-maintenance biocatalyst that is useful for detoxification of organophosphate pesticides.

Saccharomyces cerevisiae에서 Cycloinulooligosaccharide Fructanotransferase 유전자의 표층 발현 (Cell Surface Display of Cycloinulooligosaccharide Fructanotransferase Gene in Saccharomyces cerevisiae)

  • 김현진;이재형;김현철;김연희;권현주;남수완
    • 생명과학회지
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    • 제17권2호통권82호
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    • pp.241-247
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    • 2007
  • Paenibacillus macerans 유래의 cycloinulooligosaccharide fructanotransferase (CFTase) 유전자(cft, 2832 bp, 103.8 kDa)를 효모 표층발현 vector인 pCTcon (GAL1 promoter)에 subcloning 하였다. 구축된 pCTECFTN (9.0 kb)를 숙주세포인 s. cerevisiae EBY100에 형질전환한 후, uracil이 결핍된 배지와 inulin 함유배지에서 선별하였다. cft 유전자는 GAL1 promoter에 의해 효모 형질전환체에서 성공적으로 발현되었다. inulin으로부터 cyclofructans (CFs)로 생산하는 효소적 능력으로부터 표층발현 유무를 확인하였다. YPDG배지에서 48시간 배양 후 분획한 균체는 5.52 unit/ l 의 활성을 보였다. CF 생산을 위한 효소의 최적 반응 조건으로 pH 8.0, 반응온도 $50^{\circ}C$, 기질농도 5%, 기질은 Jerusalem artichoke 등의 inulin과 표층 발현 CFTase 효소반응 결과, cycloinulohexaose (CF6), cycloinuloheptaose (CF7), 그리고 cycloinulooctaose (CF8)이 생성되었고, 이 중에서 CF6가 주 생성물이었다.

효모 세포 표면 발현된 Endoxylanase를 이용한 Xylooligosaccharides의 생산 (Production of Xylooligosaccharides by Yeast Cell Surface-Displayed Endoxylanase)

  • 김현진;이재형;김연희;남수완
    • 한국미생물·생명공학회지
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    • 제36권4호
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    • pp.307-313
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    • 2008
  • Bacillus sp. endoxylanase 유전자(xynB, 642 bp)의 효모 표면발현계 pCTXYN(6.8 kb)를 구축하고 Saccharomyces cerevisiae EBY100에 형질전환시켜 형질전환체 EBY100/pCTXYN를 얻었다. 형질전환체들을 xylan이 포함된 YPDG 배지에서 배양 후 활성염색을 통하여 고찰성의 형질전환체를 최종 선별하였다. 갈락토스 배지에서 자란 효모 형질전환체로부터 xynB는 성공적으로 표면발현되었고, xylan으로 부터 xylooligosaccharides를 효율적으로 생성함도 화인하였다. Endoxylanase 활성은 세포분획에서만 검출되었고 배양 48시간에 최종 1.9 unit/mL의 활성을 보였다. Xylooligosaccharides 생산을 위한 치적 반응 조건으로, 기질과 농도는 oat spelt xylan 6%, 효모 생촉매 농도는 5 unit/mL, 반응온도는 $50^{\circ}C$, 반응시간은 $2{\sim}4$시간이었다 효모 생촉매를 oat spelt xylan과 corncob xylan에 처리한 결과, xylotriose가 주성분이었다.

Cell Surface Antigen Display for Neuronal Differentiation-Specific Tracking

  • Kim, Sang Chul;Lee, Eun-Hye;Yu, Ji Hea;Kim, Sang-Mi;Nam, Bae-Geun;Chung, Hee Yong;Kim, Yeon-Soo;Cho, Sung-Rae;Park, Chang-Hwan
    • Biomolecules & Therapeutics
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    • 제27권1호
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    • pp.78-84
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    • 2019
  • Cell therapeutic agents for treating degenerative brain diseases using neural stem cells are actively being developed. However, few systems have been developed to monitor in real time whether the transplanted neural stem cells are actually differentiated into neurons. Therefore, it is necessary to develop a technology capable of specifically monitoring neuronal differentiation in vivo. In this study, we established a system that expresses cell membrane-targeting red fluorescent protein under control of the Synapsin promoter in order to specifically monitor differentiation from neural stem cells into neurons. In order to overcome the weak expression level of the tissue-specific promoter system, the partial 5' UTR sequence of Creb was added for efficient expression of the cell surface-specific antigen. This system was able to track functional neuronal differentiation of neural stem cells transplanted in vivo, which will help improve stem cell therapies.

An integrated elastomer substrate with a lens array and pixel elements for three-dimensional liquid crystal displays

  • Hong, Jong-Ho;Kim, Yeun-Tae;Kim, Yun-Hee;Lee, Byoung-Ho;Lee, Sin-Doo
    • Journal of Information Display
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    • 제13권2호
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    • pp.55-59
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    • 2012
  • In this paper, a concept of an integrated elastomer substrate for a three-dimensional (3D) liquid crystal display based on the integral-imaging method is presented. The elemental lens array and columnar spacers were integrated into one of the two substrates, an elastomer substrate, through an imprinting process. The integrated elastomer substrate was capable of maintaining the uniform liquid crystal (LC) cell gap and promoting homeotropic LC alignment without any surface treatment. The monolithic approach reported herein will provide a key component for 3D displays with enhanced portability through a more than 40% weight reduction compared with the conventional integral-imaging method.

AC PDP(Plasma Display Panel)의 방전 특성 해석

  • 황기웅;정희섭;서정현
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 1997년도 제13회 학술발표회 논문개요집
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    • pp.173-176
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    • 1997
  • A numerical analysis of the micro-discharge in an AC pplasma dispplay cell has been made using time-deppendent, 2-dimensional multi-fluid equations to understand the discharge pphysics of He-Xe discharge. The time deppendent distribution of the electron tempperature, densities of electrons, various ions and excited sppecies, and the effects fo the wall charge accumulated on the dielectric surface are obtained and comppared with the results of direct observation of time deppendent behavior of VUV and visible sppectra from single discharge cell observed using a gated, image intensified CCD to elucidate the discharge physics.

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