biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 56:105-110, 2012 | DOI: 10.1007/s10535-012-0023-4

Biochemical properties of lipoxygenase from opium poppy chloroplasts

M. Vanko1, D. Rauová2, L. Bezáková1,*, I. Holková1, F. Bilka1, M. Cupáková2
1 Department of Cellular and Molecular Biology of Drugs, Comenius University, Faculty of Pharmacy, Bratislava, Slovak Republic
2 Toxicological and Antidoping Center, Comenius University, Faculty of Pharmacy, Bratislava, Slovak Republic

Lipoxygenase (LOX) from opium poppy (Papaver somniferum L.) chloroplasts was isolated and 126.1-fold purified to electrophoretic homogeneity by combination of ion-exchange chromatography on HA-Ultragel column and affinity chromatography on a linoleyl-aminopropyl agarose column. The relative molecular mass of the LOX determined by SDS-PAGE was 92 kDa. Kinetic properties of purified LOX were determined in spectrophotometric assay by using of linoleic acid (KM = 1.78 mM and Vmax = 11.4 μmol mg-1 min-1) and linolenic acid (KM = 1.27 mM and Vmax = 10.2 μmol mg-1 min-1). The optimum pH was 6.0 for both linoleic and linolenic acid dioxygenation catalyzed by LOX. HPLC analysis of the products revealed a dual positional specificity of linoleic acid dioxygenation at pH 6.0 with ratio of 9- and 13-hydroperoxide products being about 1:1. The activity of purified LOX was stimulated by Mg2+ and Ca2+.

Keywords: fatty acid dioxygenation; oxylipin biosynthesis; Papaver somniferum; stress signaling
Subjects: fatty acid dioxygenation; oxylipin biosynthesis; stress signaling; lipoxygenase; chloroplasts

Received: April 16, 2010; Accepted: November 4, 2010; Published: March 1, 2012  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Vanko, M., Rauová, D., Bezáková, L., Holková, I., Bilka, F., & Cupáková, M. (2012). Biochemical properties of lipoxygenase from opium poppy chloroplasts. Biologia plantarum, 56(1), 105-110. doi: 10.1007/s10535-012-0023-4
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Allen, J.C., Eriksson, C., Galpin, J.R.: Affinity chromatography of lipoxygenases. - Eur. J. Biochem. 73: 171-177, 1977. Go to original source...
    2. Andersen, M.D., Møller, B.L.: Double Triton X-114 phase partitioning for the purification of plant cytochromes P450 and removal of green pigments. - Protein Expression Purif. 13: 366-372, 1998. Go to original source...
    3. Blum, H., Beier, H., Gross, H.J.: Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. - Electrophoresis 8: 93-99, 1987. Go to original source...
    4. Bowsher, C.G., Ferrie, B.J.M., Ghosh, S., Todd, G.J., Thompson, J.E., Rothstein, S.J.: Purification and partial characterization of a membrane associated lipoxygenase in tomato fruit. - Plant Physiol. 100: 1802-1807, 1992. Go to original source...
    5. Daglia, M., Aceti, C., Giorgetti, S., Papetti, A., Gazzani, G.: Purification and characterization of soluble Cichorium intybus var. silvestre lipoxygenase. - J. Agr. Food Chem. 53: 6448-6454, 2005. Go to original source...
    6. Feussner, I., Wasternack, C.: The lipoxygenase pathway. - Annu. Rev. Plant Biol. 53: 275-297, 2002. Go to original source...
    7. Fornarolli, S., Petrussa, E., Braidot, E., Vianello, A., Macri, F.: Purification of a plasma membrane-bound lipoxygenase from soybean cotyledons. - Plant Sci. 145: 1-10, 1999. Go to original source...
    8. Garavito, R.M., Ferguson-Miller, S.: Detergents as tools in membrane biochemistry. - J. biol. Chem. 276: 32403-32406, 2001. Go to original source...
    9. Garbe, L.-A., De Almeida, R.B., Nagel, R., Wackerbauer, K., Tressl, R.: Dual positional and stereospecificity of lipoxygenase isoenzymes from germinating barley (Green Malt): biotransformation of free and esterified linoleic acid. - J. Agri. Food. Chem. 54: 946-955, 2006. Go to original source...
    10. Hornung, E., Kunze, S., Liavonchanka, A., Zimmermann, G., Kühn, D., Fritsche, K., Renz, A., Kühn, H., Feussner, I.: Identification of an amino acid determinant of pH regiospecificity in a seed lipoxygenase of Momordica charantia. - Phytochemistry 69: 2774-2780, 2008. Go to original source...
    11. Hughes, R.K., West, S.I., Hornostaj, A.R., Lawson, D.M., Fairhurst, S.A., Sanchez, R.O., Hough, P., Robinson, B.H., Casey, R.: Probing a novel potato lipoxygenase with dual positional specificity reveals primary determinants of substrate binding and requirements for a surface hydrophobic loop and has implications for the role of lipoxygenases in tubers. - Biochem. J. 15: 345-355, 2001. Go to original source...
    12. Kim, E.S., Kim, H., Park, R.D., Lee, Y., Han, O.: Dual positional specificity of wound-responsive lipoxygenase from maize seedlings. - J. Plant. Physiol. 159: 1263-1265, 2002. Go to original source...
    13. Kuo, J.M., Hwang, A., Yeh, D.B., Pan, M.H., Tsai, M.L., Pan, B.S.: Lipoxygenase from banana leaf. Purification and characterization of an enzyme that catalyzes linoleic acid oxygenation at the 9-position. - J. Agr. Food Chem. 54: 3151-3156, 2006. Go to original source...
    14. Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. - Nature 227: 680-685, 1970. Go to original source...
    15. Lang, I., Feussner, I.: Oxylipin formation in Nostoc punctiforme (PCC73102). - Phytochemistry 68: 1120-1127, 2007. Go to original source...
    16. López-Nicolas, J.M., Pérez-Gilabert, M., García-Carmona, F.: Eggplant lipoxygenase (Solanum melongena): product characterization and effect of physicochemical properties of linoleic acid on the enzymatic activity. - J. Agr. Food Chem. 49: 433-438, 2001. Go to original source...
    17. Lorenzi, V., Maury, J., Casanova, J., Berti, L.: Purification, product characterization and kinetic properties of lipoxygenase from olive fruit (Olea europaea L.). - Plant Physiol. Biochem. 44: 450-454, 2006. Go to original source...
    18. Palmieri-Thiers, C., Canaan, S., Brunini, V., Lorenzi, V., Tomi, F., Desseyn, J.L., Garscha, U., Oliw, E.H., Berti, L., Maury, J.: A lipoxygenase with dual positional specificity is expressed in olives (Olea europaea L.) during ripening. - Biochim. biophys. Acta 1791: 339-346, 2009. Go to original source...
    19. Pérez-Gilabert, M, López-Nicolás, J.M., García-Carmona, F.: Purification of a novel lipoxygenase from eggplant (Solanum melongena) chloroplasts. - Physiol. Plant. 111: 276-282, 2001. Go to original source...
    20. Porra, R.J.: The chequered history of the development and use of simultaneous equations for the accurate determination of chlorophylls a and b. - Photosynth. Res. 73: 149-156, 2002. Go to original source...
    21. Portis, A.R., Heldt, H.W.: Light-dependent changes of the Mg2+ concentration in the stroma in relation to the Mg2+ dependency of CO2 fixation in intact chloroplasts. - Biochim. biophys. Acta 449: 434-436, 1976. Go to original source...
    22. Purkait, M.K., Das Gupta, S., De, S.: Determination of thermodynamic parameters for the cloud point extraction of different dyes using TX-100 and TX-114. - Desalination 244: 130-138, 2009. Go to original source...
    23. Rangel, M., Machado, O.L.T., Da Cunha, M., Jacinto, T.: Accumulation of chloroplast-targeted lipoxygenase in passion fruit leaves in response to methyl jasmonate. - Phytochemistry 60: 619-625, 2002. Go to original source...
    24. Wang, R., Shen, W., Liu, L., Jiang, L., Liu, Y., Su, N., Wan, J.: A novel lipoxygenase gene from developing rice seeds confers dual position specificity and responds to wounding and insect attack. - Plant mol. Biol. 66: 401-414, 2008. Go to original source...

    Return to the content