Photosynthetica 2010, 48(1):3-8 | DOI: 10.1007/s11099-010-0002-y

The peroxidase activity of cytochrome b6f complex from spinach chloroplasts

X. B. Chen1, C. Hou1, L. B. Li2,*, T. Y. Kuang2,*
1 College of Bioscience and Bioengineering, Hebei University of Science and Technology, Shijiazhuang, China
2 Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing, China

The cytochrome b 6 f (Cyt b 6 f) complex, which functions as a plastoquinol-plastocyanin oxidoreductase and mediates the linear electron flow between photosystem II (PSII) and photosystem I (PSI) and the cyclic electron flow around PSI, was isolated from spinach (Spinacia oleracea L.) chloroplasts using n-octyl-β-D-glucopyranoside (β-OG). The preparation was also able to catalyze the peroxidase-like reaction in the presence of hydrogen peroxide (H2O2) and guaiacol. The optimal conditions for peroxidase activity of the preparation included: pH 3.6, ionic strength 0.1, and temperature 35°C. The apparent Michaelis constant (K m) values for H2O2 and guaiacol were 50 mM and 2 mM, respectively. The bimolecular rate constant (k obs) was about 26 M-1 s-1 and the turnover number (K cat) was about 60 min-1 (20 mM guaiacol, 100 mM sodium phosphate, pH 3.6, 25°C, [H2O2]<100mM). These parameters were similar to those of several other heme-containing proteins, such as myoglobin and Cyt c.

Keywords: cytochrome b6f complex; guaiacol; heme-containing protein; hydrogen peroxide; peroxidase activity

Received: June 24, 2009; Accepted: January 29, 2010; Published: March 1, 2010Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
Chen, X.B., Hou, C., Li, L.B., & Kuang, T.Y. (2010). The peroxidase activity of cytochrome b6f complex from spinach chloroplasts. Photosynthetica, 48(1), 3-8. doi: 10.1007/s11099-010-0002-y.
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. Baldwin, D.A., Marques, H.M., Pratt, J.M.: Hemes and hemoproteins. 5. Kinetics of the peroxidatic activity of microperoxidase-8 - model for the peroxidase enzymes. - J. Inorg. Biochem. 30: 203-217, 1987. Go to original source...
    2. Black, M.T., Widger, W.R., Cramer, W.A.: Large-scale purification of active cytochrome b 6/f complex from spinach chloroplasts. - Arch. Biochem. Biophys. 252: 655-661, 1987. Go to original source...
    3. Chen, X.B., Zhao, X.H., Zhu, Y., Gong, Y.D., Li, L.B., Zhang, J.P., Kuang, T.Y.: Hydrogen peroxide-induced chlorophyll ableaching in the cytochrome b 6 f complex: a simple and effective assay for stability of the complex in detergent solutions. - Photosynth. Res. 90: 205-214, 2006.
    4. Civello, P.M., Martínez, G.A., Chaves, A.R., Añón, M.C.: Peroxidase from strawberry fruit (Fragaria ananassa Duch) - partial-purification and determination of some properties. - J. Agr. Food Chem. 43: 2596-2601, 1995. Go to original source...
    5. Diederix, R.E.M., Ubbink, M., Canters, G.W.: The peroxidase activity of cytochrome c-550 from Paracoccus versutus. - Eur. J. Biochem. 268: 4207-4216, 2001. Go to original source...
    6. Dietrich, J., Kűhlbrandt, W.: Purification and two-dimensional crystallization of highly active cytochrome b 6 f complex from spinach. - FEBS Lett. 463: 97-102, 1999. Go to original source...
    7. Hamachi, I., Fujita, A., Kunitake, T.: Protein engineering using molecular assembly: Functional conversion of cytochrome c via noncovalent interactions. - J. Amer. Chem. Soc. 119: 9096-9102, 1997. Go to original source...
    8. Hayashi, T., Hitomi, Y., Ando, T., Mizutani, T., Hisaeda, Y., Kitagawa, S., Ogoshi, H.: Peroxidase activity of myoglobin is enhanced by chemical mutation of heme-propionates. - J. Amer. Chem. Soc. 121: 7747-7750, 1999. Go to original source...
    9. Hildebrand, D.P., Lim, K.T., Rosell, F.I., Twitchett, M.B., Wan, L., Mauk, A.G.: Spectroscopic and functional studies of a novel quadruple myoglobin variant with increased peroxidase activity. - J. Inorg. Biochem. 70: 11-16, 1998. Go to original source...
    10. Hope, A.B.: The chloroplasts cytochrome bf complex. A critical focus on function. - Biochim. Biophys. Acta 1143: 1-22, 1993. Go to original source...
    11. Hurt, E., Hauska, G.: A cytochrome f/b 6 complex of 5 polypetides with plastoquinol-plastocyanin-oxidoreductase activity from spinach chloroplasts. - Eur. J. Biochem. 117: 591-599, 1981.
    12. Kurisu, G., Zhang, H.M., Smith, J.L., Cramer, W.A.: Structure of the cytochrome b 6 f complex of oxygenic photosynthesis: Tuning the cavity. - Science 302: 1009-1014, 2003. Go to original source...
    13. 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...
    14. Lemaire, C., Girard-Bascou, J., Wollman, F.A., Bennoun, P.: Studies on the cytochrome b 6/f complex. I: Characterization of the complex subunits in Chlamydomonas reinhardtii. -Biochim. Biophys. Acta 851: 229-238, 1986. Go to original source...
    15. Mao, D.Z., Yan, J.S., Zhai, X.J., Sun, Q.M., Li, L.B.: [A new method of purifying cyt b 6 f protein complex.] - Acta Bot. Sin. 40: 1022-1027, 1998. [In Chin.]
    16. Mazumdar, S., Springs, S. L., McLendon, G. L.: Effect of redox potential of the heme on the peroxidase activity of cytochrome b 562. - Biophys. Chem. 105: 263-268, 2003. Go to original source...
    17. Nelson, D.P., Kiesow, L.A.: Enthalpy of decomposition of hydrogen peroxide by catalase at 25°C (with molar extinction coefficients of H2O2 solutions in the UV). - Anal. Biochem. 49: 474-478, 1972. Go to original source...
    18. Orii, Y.: The cytochrome c peroxidase activity of cytochrome oxidase. - J. Biol. Chem. 257: 9246-9248, 1982.
    19. Prasad, S., Maiti, N.C., Mazumdar, S., Mitra, S.: Reaction of hydrogen peroxide and peroxidase activity in carboxyme thylated cytochrome c: spectroscopic and kinetic studies. - Biochim. Biophys. Acta 1596: 63-75, 2002. Go to original source...
    20. Radi, R., Thomson, L., Rubbo, H., Prodanov, E.: Cytochrome c catalyzed oxidation of organic molecules by hydrogen peroxide. - Arch. Biochem. Biophys. 288: 112-117, 1991. Go to original source...
    21. Rosei, M.A., Blarzino, C., Coccia, R., Foppoli, C., Mosca, L., Cini, C.: Production of melanin pigments by cytochrome c/H2O2 system. - Int. J. Biochem. Cell Biol. 30: 457-463, 1998. Go to original source...
    22. Savenkova, M. I., Kuo, J. M., Ortiz de Montellano, P. R.: Improvement of peroxygenase activity by relocation of a catalytic histidine within the active site of horseradish peroxidase. - Biochemistry 37: 10828-10836, 1998. Go to original source...
    23. Stroebel, D., Choquet, Y., Popot, J.L., Picot, D.: An atypical haem in the cytochrome b 6 f complex. - Nature 426: 413-418, 2003. Go to original source...
    24. Thomas, P.E., Ryan, D., Levin, W.: An improved staining procedure for the detection of the peroxidase activity of cytochrome P-450 on sodium dodecyl sulfate polyacylamide gels. - Anal. Biochem. 75: 168-176, 1976. Go to original source...
    25. Vácha, F., Vácha, M., Bumba, L., Hashizume, K., Tani, T.: Inner structure of intact chloroplasts observed by a low temperature laser scanning microscope. - Photosynthetica 38: 493-496, 2000. Go to original source...
    26. Vazquez-Duhalt, R.: Cytochrome c as a biocatalyst. - J. Mol. Catal. B: Enzym. 7: 241-249, 1999. Go to original source...
    27. Yonetani, T.: Cytochrome c peroxidase (Brkers' yeast). - Methods Enzymol. 10: 336-339, 1976. Go to original source...
    28. Yan, J.S., Liu, Y.L., Mao, D.Z., Li, L.B., Kuang, T.Y.: The presence of 9-cis-β-carotene in cytochrome b 6 f complex from spinach. - Biochim. Biophys. Acta: 1506: 182-188, 2001. Go to original source...

    Return to the content