UV Spectrometric Assay of Epoxide Hydrolase Activity of Microbial Cell Biocatalysts

자외선분광기를 이용한 미생물 세포 생촉매의 에폭사이드 가수분해효소 활성평가

  • Kim, Hee Sook (Department of Food Science and Technology, Kyungsung University) ;
  • Lee, Eun Yeol (Department of Food Science and Technology, Kyungsung University)
  • Received : 2005.02.02
  • Accepted : 2005.03.31
  • Published : 2005.06.10

Abstract

UV spectrometric assay for measurement of epoxide hydrolase activity was tested for efficient screening of whole cell activity of epoxide hydrolase. Epoxide hydrolase activities were determined by measuring the amount of p-nitrostyrene diol (pNSD), which was the hydrolysis product of p-nitrostyrene oxide (pNSO). Enantioselective hydrolysis of racemic pNSO using epoxide hydrolase activity of Rhodosporidium toruloides was monitored by UV spectrometric assay, and the relevant $K_m$ and $V_m$ for R. toruloides were determined as $2.457nmol/min{\cdot}mg$ and 1.078 mM, respectively.

미생물 세포 유래의 에폭사이드 가수분해 효소 활성을 효율적으로 분석하기 위하여 UV 분광기 측정법을 최적화하였다. 세포 생촉매의 에폭사이드 가수분해 활성에 의해 p-nitrostyrene oxide (pNSO) 기질이 p-nitrostyrene diol로 전환된 양을 측정함으로써 에폭사이드 가수분해 활성을 평가하였다. Rhodosporidium toruloides를 생촉매로 사용하고 pNSO에 대한 입체선택적 가수분해 동력학을 UV 분광기 측정법으로 분석한 결과, $K_m$$V_m$ 값을 $2.457nmol/min{\cdot}mg$ 및 1.078 mM로 각각 결정할 수 있었다.

Keywords

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