Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Development of Recombinant Saccharomyces cerevisiae Expressing Epoxide Hydrolase for the Preparation of Chiral Phenyl Oxirane

광학활성 Phenyl Oxirane 제조용 유전자 재조합 생촉매 개발

  • 이수정 (경성대학교 공과대학 식품공학과) ;
  • 이은정 (경성대학교 공과대학 식품공학과) ;
  • 김초희 (경성대학교 공과대학 식품공학과) ;
  • 이지원 (경성대학교 공과대학 식품공학과) ;
  • 김희숙 (경성대학교 공과대학 식품공학과) ;
  • 이은열 (경성대학교 공과대학 식품공학과)
  • Published : 2003.02.01
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Abstract

Epoxide hydrolase (EH) gene from Aspergillus niger #33 was cloned from cDNA library generated by reverse transcriptase-polymerase chain reaction (RT-PCR). The nucleotide sequence analysis revealed that the deduced amino acid sequence was almost similiar to that of A. niger LCP521 previously reported. The cloned EH gene was transformed into Saccharomyces cerevisiae and expressed by addition of galactose. The recombinant S. cerevisiae showed hydrolytic activity toward racemic phenyl oxirane substrate based on chiral GC analysis, and can be used as a potential biocatalyst for the preparation of chiral phenyl oxirane.

Aspergillus niger #33로부터 RT-PCR 및 PCR방법을 이용하여 epoxide hydrolase (EH)유전자를 클로닝 하고 염기서 열을 분석한 결과 A. niger LCP521 유래의 EB와 85%수준의 유사성을 가지고 있었다. 클로닝된 EH 유전자를 Sacrharomyces cerevisiae에 형질전환 시킨 후 galactose를 inducer로 사용하여 발현시켰다. 유전자 재조합 S. cerevisiae는 라세믹 phenyl oxirane 기질에 대하여 입체선택적 가수분해능이 있음을 확인할 수 있었으며, 이러한 유전자 재조합 EH는 광학활성 에폭사이드 제조를 위한 생촉매로 응용될 수 있을 것이다.

Keywords

References

  1. Biochem. J. v.337 Catalytic triad of microsomal epoxide hydrolase: replacement of Glu404 with Asp leads to a strongly increased turnover rate Arand, M.;F. Muller;A. Mecky;W. Hinz;P. Urban;D. Pompon;R. Kellner;F. Oesch https://doi.org/10.1042/0264-6021:3370037
  2. Biochem. J. v.344 Cloning and molecular characterization of a soluble epoxide hydrolase from Aspergillus niger that is related to mammalian microsomal epoxide hydrolase Arand, M.;H. Hemmer;H. Durk;J. Baratti;A. Archelas;R. Furstoss https://doi.org/10.1042/0264-6021:3440273
  3. J. Biol. Chem. v.275 Binding of alkylurea inhibitors to epoxide hydrolase implicates active site tyrosines in substrate activation Argiriadi, M.A,;C. Morisseau;M. H. Goodrow;D. L. Dowdy;B. D. Hammock;D. W. Christianson https://doi.org/10.1074/jbc.M000278200
  4. Tetrahedron v.50 Chemical and biological synthesis of chiral epoxides Besse, P.;H. Veschambre https://doi.org/10.1016/S0040-4020(01)85362-X
  5. Anal. Biochem. v.162 Single-step mithod of RNA isolation by acid guanidinum thiocyante-phenol-chloroform extraction Chomczynski, P.;N. Szcchi
  6. J. Biol. Chem. v.274 The X-ray structure of epoxide hydrolase from Agrobacterium radiobacter AD1 Nardini, M.;I. S. Ridder;H. J. Rozeboom;K. H. Kalk;R. Rink;D. B. Janssen;B. W. Dijkstra https://doi.org/10.1074/jbc.274.21.14579
  7. J. Am. Chem. Soc. v.121 Mutation fo tyrosine residues involved in the alkylation half reaction of epoxide hydrolase from Agrobacterium radiobacter AD1 results in improved enantioselectivity Rink, R.;J. H. L. Spelberg;R. J. Pieters;J. Kingma;M. Nardini;R. M. Kellogg;B. W. Dijkstra;D. B. Janssen https://doi.org/10.1021/ja990501o
  8. Current Opinion in Biotechnol. v.12 Microbial epoxide hydrolases for preparative biotransformations Steinreiber, A.;K. Faber https://doi.org/10.1016/S0958-1669(01)00262-2
  9. Appl. Environ. Microbiol. v.65 Isolation and characterization fo the epoxide hydrolase-encoding gene from Xanthophyllomyces dendrorhous Visser, H.;J. A. M. de Bont;J. C. Verdoes
  10. J. Mol. Catal. B.: Enzym. v.6 Epoxide hydrolases from yeasts and other sources: versatile tools in biocatalysis Weijers, C.A. G. M.;J. A. M. de Bont https://doi.org/10.1016/S1381-1177(98)00123-4
  11. J. Biol. Chem. v.275 Biochemical evidence for the involvement of tyrosine in epoxide activation during the catalytic cycle of epoxide hydrolase Yamada, Y.;C. Morisseau;J. E. Maxwell;M. A. Argiriadi;D. W. Christianson;B. D. Hammock https://doi.org/10.1074/jbc.M001464200
  12. Structure v.8 Structure of Aspergillus niger epoxide hydrolase at 1.8Å resolution: implication for the structure and function of the mammalian microsomal class of epoxide hydrolases Zou, J.;B. M. Hallberg;T. Bergfors;F. Oesch;M. Arand;S. L. Mowbray;T. A. Jones https://doi.org/10.1016/S0969-2126(00)00087-3

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