자외선분광기를 이용한 미생물 세포 생촉매의 에폭사이드 가수분해효소 활성평가

김희숙; 이은열; Hee Sook Kim; Eun Yeol Lee

Journal of the Korean Industrial and Engineering Chemistry, Vol.16, No.3, 456-459, June, 2005

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자외선분광기를 이용한 미생물 세포 생촉매의 에폭사이드 가수분해효소 활성평가

UV Spectrometric Assay of Epoxide Hydrolase Activity of Microbial Cell Biocatalysts

김희숙, 이은열 ^†

경성대학교 식품공학과

Hee Sook Kim, and Eun Yeol Lee^†

Department of Food Science and Technology, Kyungsung Univesity, Busan 608-736, Korea

E-mail:

초록

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, Km과 Vm 값을 2.457 nmol/minㆍmg 및 1.078 mM로 각각 결정할 수 있었다.

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 Km and Vm for R. toruloides were determined as 2.457 nmol/minㆍmg and 1.078 mM, respectively.

Keywords:epoxide hydrolase;p-nitrostyrene diol;p-nitrostyrene oxide;Rhodosporidium toruloides;spectrometric assay

[References]

Kasai N, Suzuki T, Furukawa Y, J. Mol. Catal. B-Enzym., 4, 237 (1998)
deVries EJ, Janssen DB, Curr. Opin. Biotechnol., 14, 414 (2003)
Lee JH, Lee EY, J. Korean Ind. Eng. Chem., 15(1), 150 (2004)
Zocher F, Enzelberger MM, Bornscheuer UT, Hauer B, Schmid RD, Anal. Chim. Acta, 391, 345 (1999)
Mateo C, Archelas A, Furstoss R, Anal. Biochem., 314, 135 (2003)
Dodere K, Lutz-Wahl S, Hauer B, Schmid RD, Anal. Biochem., 321, 131 (2003)
Bhatnagar T, Manoj KM, Baratti C, J. Biochem. Biophys. Methods, 50, 1 (2001)
Lee EY, Yoo SS, Kim HS, Lee SJ, Oh YK, Park S, Enzyme Microb. Technol., 35(6-7), 624 (2004)
Westkaemper RB, Hanzlik RP, Anal. Biochem., 102, 63 (1980)
Choi WJ, Lee EY, Yoon SJ, Yang ST, Choi CY, J. Biosci. Bioeng., 88(3), 339 (1999)

[Related Articles]

[1] Kasai N, Suzuki T, Furukawa Y, J. Mol. Catal. B-Enzym., 4, 237 (1998)

[2] deVries EJ, Janssen DB, Curr. Opin. Biotechnol., 14, 414 (2003)

[3] Lee JH, Lee EY, J. Korean Ind. Eng. Chem., 15(1), 150 (2004)

[4] Zocher F, Enzelberger MM, Bornscheuer UT, Hauer B, Schmid RD, Anal. Chim. Acta, 391, 345 (1999)

[5] Mateo C, Archelas A, Furstoss R, Anal. Biochem., 314, 135 (2003)

[6] Dodere K, Lutz-Wahl S, Hauer B, Schmid RD, Anal. Biochem., 321, 131 (2003)

[7] Bhatnagar T, Manoj KM, Baratti C, J. Biochem. Biophys. Methods, 50, 1 (2001)

[8] Lee EY, Yoo SS, Kim HS, Lee SJ, Oh YK, Park S, Enzyme Microb. Technol., 35(6-7), 624 (2004)

[9] Westkaemper RB, Hanzlik RP, Anal. Biochem., 102, 63 (1980)

[10] Choi WJ, Lee EY, Yoon SJ, Yang ST, Choi CY, J. Biosci. Bioeng., 88(3), 339 (1999)