Biochemistry (Moscow)

, Volume 75, Issue 7, pp 861–865 | Cite as

Recombinant maize 9-lipoxygenase: Expression, purification, and properties

  • E. V. Osipova
  • I. R. Chechetkin
  • Y. V. Gogolev
  • N. B. TarasovaEmail author


Expression of maize 9-lipoxygenase was performed and optimized in Escherichia coli Rosetta(DE3)pLysS. The purity of recombinant protein obtained during Q-Sepharose and Octyl-Sepharose chromatographies in an LP system at 4°C was >95%. Maximum activity of the lipoxygenase reaction was observed at pH 7.5. Enzyme stability was studied at pH 4.5 to 9.5 and in the presence of different compounds: phenylmethanesulfonyl fluoride, β-mercaptoethanol, ammonium sulfate, and glycerol. HPLC and GC-MS analysis showed that enzyme produced 99% 9S-hydroperoxide from linoleic acid. 13-Hydroperoxide (less than 1%) consisted of S- and R-enantiomers in ratio 2 : 3.

Key words

maize 9-lipoxygenase expression of 9-lipoxygenase chromatography of 9-lipoxygenase GC-MS analysis HPLC 



gas chromatography-mass spectrometry


high performance liquid chromatography


isopropyl β-D-1-thiogalactopyranoside


Luria-Bertani (medium)


phenylmethanesulfonyl fluoride


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Copyright information

© Pleiades Publishing, Ltd. 2010

Authors and Affiliations

  • E. V. Osipova
    • 1
  • I. R. Chechetkin
    • 1
  • Y. V. Gogolev
    • 1
  • N. B. Tarasova
    • 1
    Email author
  1. 1.Kazan Institute of Biochemistry and Biophysics, Kazan Research CenterRussian Academy of SciencesKazanRussia

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