Recombinant maize 9-lipoxygenase: Expression,purification, and properties |
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| Authors: | E. V. Osipova I. R. Chechetkin Y. V. Gogolev N. B. Tarasova |
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| Affiliation: | 1.Kazan Institute of Biochemistry and Biophysics, Kazan Research Center,Russian Academy of Sciences,Kazan,Russia |
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| Abstract: | 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. |
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