DOI QR코드

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재조합 박테리아 바이오센서의 고정화: 페놀계 화합물의 현장 검출을 위한 간단한 접근 방법

Immobilization of Recombinant Bacterial Biosensors: a Simple Approach for the On-Site Detection of Phenolic Compounds

  • 신혜자 (동서대학교 에너지생명공학부 에너지환경공학)
  • Shin, Hae-Ja (Energy Environmental Engineering Major, Division of Energy Bioengineering, Dongseo University)
  • 투고 : 2011.08.01
  • 심사 : 2011.09.05
  • 발행 : 2011.09.30

초록

본 연구에서는 페놀 화합물들을 현장에서 검출하기 위해 간단하고 간편한 일회용 재조합 박테리아 바이오센서 시스템을 개발하였다. 플라즈미드 pLZCapR을 함유하는 E. coli 세포를 ${\beta}$-galactosidase 기질인 CPRG와 함께 96-well plate의 wells에 agarose로 고정하였다. 이 바이오센서는 현장에서 발색을 위한 별도의 기질을 추가하거나 불편한 기기 사용 또는 시료의 전 처리를 필요로 하지 않는다. 시료의 측정은 간단히 적은 부피(<100 ${\mu}l$)의 현장 시료를 바이오센서를 포함하는 wells에 넣고 발색을 관찰하여 측정하였다. 또한 6% DMF, 0.1% SDS 그리고 10 mM $CaCl_2$를 첨가하여 agarose 고정에 의한 화합물의 세포내 확산 제한을 감소시켜 보다 더 나은 발색을 얻을 수 있었다. 따라서 이 고정된 미생물 유래 재조합 바이오센서 시스템은 현장에서 환경오염물질들을 간단하게 확인하고 정량 하는 유용한 접근 방법이 될 것으로 사료된다.

We herein report the development of an agarose-gel-immobilized recombinant bacterial biosensor simple system for the field monitoring of phenolic compounds. Escherichia coli cells harboring the pLZCapR plasmid, which was previously designed to express the ${\beta}$-galactosidase reporter gene in the presence of phenolic compounds, were co-immobilized with a substrate [chlorophenol red ${\beta}$-galactopyranoside (CPRG) in agarose gel, and dispensed to the wells of a 96-well plate. Field samples were added to the wells and color development was monitored. In the presence of 5 ${\mu}M$ to 10 mM of phenol, the biosensor developed a red (representing hydrolysis of CPRG) color. Other phenolic compounds were also detected by this immobilized system, with the pattern resembling that previously reported for the corresponding non-immobilized biosensor. The immobilized cells showed optimum activity when the gel was simultaneously supplemented with 6% dimethyl formamide (DMF), 0.1% SDS and 10 mM $CaCl_2$. The immobilized biosensor described herein does not require the addition of a substrate or the use of unwieldy instruments or sample pretreatments that could complicate field studies.

키워드

참고문헌

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