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)
  • 김희숙 (경성대학교 식품공학과) ;
  • 이은열 (경성대학교 식품공학과)
  • 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.

References

  1. N. Kasai, T. Suzuki, and Y Furukawa, J. Mol. Cat. B: Enz., 4, 237 (1998) https://doi.org/10.1016/S1381-1177(97)00034-9
  2. E. J. de Vries and D. B. Janssen, Curr. Opin. Biotechnol., 14, 414 (2003) https://doi.org/10.1016/S0958-1669(03)00102-2
  3. J. H. Lee and E. Y. Lee, J. Korean Ind. Eng. Chem., 15, 150 (2004)
  4. F. Zocher, M. M. Enze1berger, U. T. Bornscheuer, B. Hauer, and R. D. Schmid, Anal. Chim. Acta, 391, 345 (1999) https://doi.org/10.1016/S0003-2670(99)00216-0
  5. C. Mateo, A. Archelas, and R. Furstoss, Anal. Biochem., 314, 135 (2003) https://doi.org/10.1016/S0003-2697(02)00646-2
  6. K. Dodere, S. Lutz-Wahl, B. Hauer, and R. D. Schmid, Anal. Biochem., 321, 131 (2003) https://doi.org/10.1016/S0003-2697(03)00399-3
  7. T. Bhatnagar, K. M. Manoj, and C. Baratti, J. Biochem. Biophys. Methods, 50, 1 (2001) https://doi.org/10.1016/S0165-022X(01)00162-2
  8. E. Y. Lee, S.-S. Yoo, H. S. Kim, S. J. Lee, Y.-K. Oh, and S. Park, Enzym. Microbial Technol., 35, 624 (2004)
  9. R. B. Westkaemper and R P. Hanzlik, Anal. Biochem., 102, 63 (1980) https://doi.org/10.1016/0003-2697(80)90317-6
  10. W. J. Choi, E. Y. Lee, S. J. Yoon, and C. Y. Choi, J. Biosci. Bioeng., 88, 339 (1999) https://doi.org/10.1016/S1389-1723(00)80022-5