Aspergillus niger의 Epoxide Hydrolase 고효율 발현 및 라세믹 에폭사이드의 입체선택적 가수분해

이수정; 김희숙; 이은열; Soo Jung Lee; Hee Sook Kim; Eun Yeol Lee

Journal of the Korean Industrial and Engineering Chemistry, Vol.17, No.5, 557-560, October, 2006

Enhanced Heterologous Expression of Aspergillus niger Epoxide Hydrolase and Its Application to Enantioselective Hydrolysis of Racemic Epoxides

이수정, 김희숙, 이은열 ^†

경성대학교 공과대학 식품공학과

Soo Jung Lee, Hee Sook Kim, and Eun Yeol Lee^†

Department of Food Science and Technology, College of Engineering, Kyungsung University, Busan 608-736, Korea

E-mail:

초록

Aspergillus niger LK의 epoxide hydrolase (EH)를 codon usage를 고려한 Escherichia coli 균주에서 고효율로 발현할 수 있었다. E. coli에서 잘 사용되지 않는 rare codon에 대한 tRNA 유전자 정보가 들어있는 plasmid를 함유한 E. coli 균주인 Rosetta (DE3)PLysS를 숙주세포로 사용하였다. A. niger EH를 발현시킨 재조합 E. coli를 생촉매로 사용하여 라세믹 styrene oxide 혼합물과 반응시켰을 때, (R)-styrene oxide에 대한 입체선택적 가수분해활성이 향상됨을 확인할 수 있었다. 또한 라세믹 기질로부터 입체적으로 고순도인 99% ee 값을 갖는 광학적으로 순수한 (S)-styrene oxide를 얻을 수 있었다.

The epoxide hydrolase (EH) of Aspergillus niger LK was expressed to high levels in Escherichia coli based on codon usage. E. coli, Rosetta (DE3)PLysS, containing a large number of tRNAs for rare-codons was employed as a host strain. The recombinant E. coli expressing A. niger EH showed an enhanced enantioselective hydrolysis activity toward racemic styrene oxide. Enantiopure (S)-styrene oxide with a high enantiopurity of 99% ee was obtained from racemic substrates.

Keywords:epoxide hydrolase;Aspergillus niger;enantioselective hydrolysis;Escherichia coli Rosetta;codon usage

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Steinreiber A, Faber K, Curr. Opin. Chem. Biol., 12, 552 (2001)
Weijers CAGM, de Bont JAM, J. Mol. Catal. B-Enzym., 6, 199 (1999)
Lee EY, Kor. J. Biotechnol. Bioeng, 17, 321 (2002)
Besse P, Veschambre H, Tetrahedron, 50, 8885 (1994)
Arand M, Muller F, Mecky A, Hinz W, Urban P, Pompon D, Kellner R, Oesch F, Biochem. J., 337, 37 (1999)
Nardini M, Ridder IS, Rozeboom HJ, Kalk KH, Rink R, Janssen DB, Dijkstra BW, J. Biol. Chem., 274, 14579 (1999)
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Lee EY, Kim HS, Kor. J. Biotechnol. Bioeng., 16, 562 (2001)
Visser H, Vreugdenhil S, de Bont JAM, Verdoes JC, Appl. Microbiol. Biotechnol, 53, 415 (2000)
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[Related Articles]

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

[2] Steinreiber A, Faber K, Curr. Opin. Chem. Biol., 12, 552 (2001)

[3] Weijers CAGM, de Bont JAM, J. Mol. Catal. B-Enzym., 6, 199 (1999)

[4] Lee EY, Kor. J. Biotechnol. Bioeng, 17, 321 (2002)

[5] Besse P, Veschambre H, Tetrahedron, 50, 8885 (1994)

[6] Arand M, Muller F, Mecky A, Hinz W, Urban P, Pompon D, Kellner R, Oesch F, Biochem. J., 337, 37 (1999)

[7] Nardini M, Ridder IS, Rozeboom HJ, Kalk KH, Rink R, Janssen DB, Dijkstra BW, J. Biol. Chem., 274, 14579 (1999)

[8] Visser H, de Bont JAM, Verdoes JC, Appl. Environ. Microbiol., 65, 5459 (1999)

[9] Arand M, Hemmer H, Durk H, Baratti J, Archelas A, Furstoss R, Biochem. J., 344, 273 (1999)

[10] Argiriadi MA, Morisseau C, Goodrow MH, Dowdy DL, Hammock BD, Christianson DW, J. Biol. Chem., 275, 15265 (2000)

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

[12] Yamada T, Morisseau C, Maxwell JE, Argiriadi MA, Christianson DW, Hammock BD, J. Biol. Chem., 275, 23082 (2000)

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

[14] Choi WJ, Huh EC, Park HJ, Lee EY, Choi CY, Biotechnol. Tech., 12, 225 (1998)

[15] Nakamura Y, Gojobori T, Ikemura T, Nucleic Acids Res., 28, 292 (2000)

[16] Lee EY, Kim HS, Kor. J. Biotechnol. Bioeng., 16, 562 (2001)

[17] Visser H, Vreugdenhil S, de Bont JAM, Verdoes JC, Appl. Microbiol. Biotechnol, 53, 415 (2000)

[18] Manoj KM, Archelas A, Baratti J, Furstoss R, Tetrahedron, 57, 695 (2001)

[19] Novy R, Drott D, Yaeger K, Mierendorf R, Newsletter of Novagen, 12, 1 (2001)