Development and characterization of recombinant whole cells expressing the soluble epoxide hydrolase of Danio rerio and its variant for enantioselective resolution of racemic styrene oxides

Mi Hee Woo; Hee Sook Kim; Eun Yeol Lee

Journal of Industrial and Engineering Chemistry, Vol.18, No.1, 384-391, January, 2012

DOI10.1016/j.jiec.2011.11.110 Export Citation

Development and characterization of recombinant whole cells expressing the soluble epoxide hydrolase of Danio rerio and its variant for enantioselective resolution of racemic styrene oxides

Mi Hee Woo, Hee Sook Kim^†, and Eun Yeol Lee^{1, †}

Department of Food Science and Biotechnology, Kyungsung University, Busan 608-736, Republic of Korea
¹Department of Chemical Engineering, Kyung Hee University, Gyeonggi-do 446-701, Republic of Korea

E-mail:,

A recombinant soluble epoxide hydrolase (sEH) of zebra fish, Danio rerio, and its variant were developed and characterized. The variant consisted of five-point mutations such as Glu88Arg, Thr102Ala, Met368Ile, Lys398Glu and Gly412Asp. The catalytically important amino acids of Asp331, Tyr379, Tyr460, Asp496 and His524 were determined to be highly conserved in both of the sEHs, on the basis of multiple sequence alignment and homology modeling. The enantiomeric ratio of the variant sEH was 1.43-fold higher than that of the wild-type sEH. Interestingly, both of the sEH and its variant possessed (S)-styrene oxide-preferred hydrolytic activity, while the microsomal EH (mEH) of D. rerio exhibited the enantiopreference toward (R)-enantiomer, indicating that (R)- and (S)-styrene oxide could be prepared by using sEH and mEH, respectively. (R)-Styrene oxide with high enantiopurity of 99%ee could be obtained by the enantioselective hydrolysis activities of the recombinant Escherichia coli whole cells expressing the wild-type and variant sEHs of D. rerio.

Keywords:Danio rerio;Epoxide hydrolase;Mutation;Chiral styrene oxide;Enantioselective hydrolysis

[References]

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Lee EY, Kim HS, Kor. J. Microbiol. Biotechnol., 34, 129 (2006)
Lee SJ, Kim HS, Kim SJ, Park S, Kim BJ, Shuler ML, Lee EY, Biotechnol. Lett., 29(2), 237 (2007)
Kim HS, Lee OK, Hwang S, Kim BJ, Lee EY, Biotechnol. Lett., 30(1), 127 (2008)
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Choi SH, Kim HS, Lee EY, Biotechnol. Lett., 10, 1617 (2009)
Barth S, Fischer M, Schmid RD, Pleiss J, Bioinformatics., 20, 2845 (2004)
Arand M, Wagner H, Oesch F, J. Biol. Chem., 271, 4223 (1996)
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[1] Hwang S, Choi CY, Lee EY, J. Ind. Eng. Chem., 16(1), 1 (2010)

[2] Orru RVA, Faber K, Curr. Opin. Chem. Biol., ., 16 (1999)

[3] Archelas A, Furstoss R, Curr. Opin. Chem. Biol., 5, 112 (2001)

[4] de Vries EJ, Janssen DB, Curr. Opin. Biotechnol., 14, 414 (2003)

[5] Gong PF, Xu JH, Tang YF, Wu HY, Biotechnol. Prog., 19(2), 652 (2003)

[6] Lee EY, Shuler ML, Biotechnol. Bioeng., 98(2), 318 (2007)

[7] Lee EY, Kim HS, Kor. J. Microbiol. Biotechnol., 34, 129 (2006)

[8] Lee SJ, Kim HS, Kim SJ, Park S, Kim BJ, Shuler ML, Lee EY, Biotechnol. Lett., 29(2), 237 (2007)

[9] Kim HS, Lee OK, Hwang S, Kim BJ, Lee EY, Biotechnol. Lett., 30(1), 127 (2008)

[10] Cedrone F, Niel S, Ait-abdelkader N, Torre C, Krumm H, Roca S, Bhatnagar T, Maichele A, Reetz MT, Baratti JC, Biocatal. Biotransform., 21, 357 (2003)

[11] Reetz MT, Torre C, Eipper A, Lohmer R, Hermes M, Brunner B, Maichele A, Bocola M, Arand M, Cronin A, Genzel Y, Archelas M, Furstoss R, Org. Lett., 6, 177 (2004)

[12] Reetz MT, Adv. Catal., 49, 1 (2006)

[13] van Loo B, Spelberg JHL, Kingma J, Sonke T, Wubbolts MG, Janssen DB, Chem.Biol., 11, 981 (2004)

[14] Schrader W, Eipper A, Pugh DJ, Reetz MT, Can. J. Chem., 80, 626 (2002)

[15] Choi SH, Kim HS, Lee EY, Biotechnol. Lett., 10, 1617 (2009)

[16] Barth S, Fischer M, Schmid RD, Pleiss J, Bioinformatics., 20, 2845 (2004)

[17] Arand M, Wagner H, Oesch F, J. Biol. Chem., 271, 4223 (1996)

[18] Zou J, Hallberg BM, Bergfors T, Oesch F, Arand M, Mowbray SL, Jones TA, Structure., 8, 111 (2000)

[19] Rink R, Spelberg JHL, Pieters RJ, Kingma J, Nardini M, Kellogg RM, Dijkstra BW, Janssen DB, J. Am. Chem. Soc., 121(32), 7417 (1999)