Biotechnology Letters

, Volume 32, Issue 11, pp 1685–1691 | Cite as

Functional expression and magnetic nanoparticle-based Immobilization of a protein-engineered marine fish epoxide hydrolase of Mugil cephalus for enantioselective hydrolysis of racemic styrene oxide

  • Sung Hee Choi
  • Hee Sook Kim
  • In Su Lee
  • Eun Yeol Lee
Original Research Paper

Abstract

A triple-point mutated fish microsomal epoxide hydrolase (mEH) gene from Mugil cephalus was expressed in Escherichia coli in the presence of various chaperones to prevent protein aggregations. The enantioselective hydrolytic activity was more than doubled by co-expressing the EH mutant gene with pGro7 plasmid. The highly active EH mutant with a his-tag was immobilized onto magnetic silica assembled with NiO nanoparticles. The immobilized mEH mutant was re-used more than 10 times with less than 10% activity loss. (S)-Styrene oxide with 98% enantiopurity was repeatedly obtained with over 50% of the theoretical yield by the magnetically separable high-performance mEH mutant.

Keywords

Chaperone Chiral epoxides Epoxide hydrolase Immobilization Mugil cephalus 

References

  1. Brady D, Jordaan J (2009) Advances in enzyme immobilization. Biotechnol Lett 31:1639–1650CrossRefPubMedGoogle Scholar
  2. Choi SH, Kim HS, Lee EY (2009) Comparative homology modeling-inspired protein engineering for improvement of catalytic activity of Mugil cephalus epoxide hydrolase. Biotechnol Lett 31:1617–1624CrossRefPubMedGoogle Scholar
  3. de Vries EJ, Janssen DB (2003) Biocatalytic conversion of epoxides. Curr Opin Biotechnol 14:414–420CrossRefPubMedGoogle Scholar
  4. Fink AL (1999) Chaperone-mediated protein folding. Physiol Rev 79:425–449PubMedGoogle Scholar
  5. Hwang S, Choi CY, Lee EY (2010) Bio- and chemo-catalytic preparation of chiral epoxides. J Ind Eng Chem 16:1–6Google Scholar
  6. Kasai N, Suzuki T, Furukawa Y (1998) Chiral C3 epoxides and halohydrins: Their preparation and synthetic application. J Mol Catal B: Enzym 4:237–252CrossRefGoogle Scholar
  7. Kim J, Grate JW, Wang P (2006) Nanostructures for enzyme stabilization. Chem Eng Sci 61:1017–1026CrossRefGoogle Scholar
  8. Lee KS, Lee IS (2008) Decoration of superparamagnetic iron oxide nanoparticles with Ni2+: agent to bind and separate histidine-tagged proteins. Chem Comm 2008:709–711CrossRefGoogle Scholar
  9. Lee SJ, Kim HS, Kim SJ, Park S, Kim BJ, Shuler ML, Lee EY (2007) Cloning, expression and enantioselective hydrolytic catalysis of a microsomal epoxide hydrolase from a marine fish, Mugil cephalus. Biotechnol Lett 29:237–246CrossRefPubMedGoogle Scholar
  10. Lee KS, Woo MH, Kim HS, Lee EY, Lee IS (2009) Synthesis of hybrid Fe3O4/silica/NiO superstructures and their application as magnetically separable high-performance biocatalysts. Chem Comm 25:3780–3782CrossRefPubMedGoogle Scholar
  11. Sørensen HP, Mortensen KK (2005) Advanced genetic strategies for recombinant protein expression in Escherichia coli. J Biotechnol 115:113–128CrossRefPubMedGoogle Scholar
  12. Steinreiber A, Faber K (2001) Microbial epoxide hydrolases for preparative biotransformations. Curr Opin Biotechnol 12:552–558CrossRefPubMedGoogle Scholar
  13. Tran TAT, Struck DK, Young R (2005) Periplasmic domains define holin-antiholin interactions in T4 lysis inhibition. J Bacteriol 187:6631–6640CrossRefPubMedGoogle Scholar
  14. Widersten M, Gurell A, Lindberg D (2010) Structure-function relationships of epoxide hydrolases and their potential use in biocatalysis. Biochim Biophys Acta 1800:316–326PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Sung Hee Choi
    • 1
  • Hee Sook Kim
    • 1
  • In Su Lee
    • 2
  • Eun Yeol Lee
    • 3
  1. 1.Department of Food Science and BiotechnologyKyungsung UniversityBusanSouth Korea
  2. 2.Department of Applied ChemistryKyung Hee UniversityGyeonggi-doKorea
  3. 3.Department of Chemical Engineering and Industrial Liaison Research InstituteKyung Hee UniversityGyeonggi-doKorea

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