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. 1982 Nov;70(5):1249–1254. doi: 10.1104/pp.70.5.1249

Hydrogen Peroxide Synthesis in Isolated Spinach Chloroplast Lamellae 1

An Analysis of the Mehler Reaction in the Presence of NADP Reduction and ATP Formation

J Michael Robinson 1,2, Martin Gibbs 1,2
PMCID: PMC1065870  PMID: 16662662

Abstract

Light-dependent O2 reduction concomitant with O2 evolution, ATP formation, and NADP reduction were determined in isolated spinach (Spinacia oleracea L. var. America) chloroplast lamellae fortified with NADP and ferredoxin. These reactions were investigated in the presence or absence of catalase, providing a tool to estimate the reduction of O2 to H2O2 (Mehler reaction) concomitant with NADP reduction. In the presence of 250 micromolar O2, O2 photoreduction, simultaneous with NADP photoreduction, was dependent upon light intensity, ferredoxin, Mn2+, NADP, and the extent of coupling of phosphorylation to electron flow.

In the presence of an uncoupling concentration of NH4+, saturating light intensity (>500 watts/square meter), saturating ferredoxin (10 micromolarity) rate-limiting to saturating NADP (0.2-0.9 millimolarity), and Mn2+ (50-1000 micromolarity), the maxium rates of O2 reduction were 13-25 micromoles/milligram chlorophyll per hour, while concomitant rates of O2 evolution and NADP reduction were 69 to 96 and 134 to 192 micromoles/milligram chlorophyll per hour, respectively. Catalase did not affect the rate of NADPH or ATP formation but decreased the NADPH:O2 ratios from 2.3-2.8 to 1.9-2.1 in the presence of rate-limiting as well as saturating concentrations of NADP.

Photosynthetic electron flow at a rate of 31 micromoles O2 evolved/milligram chlorophyll per hour was coupled to the synthesis of 91 micromoles ATP/milligram chlorophyll per hour, while the concomitant rate of O2 reduction was 0.6 micromoles/milligram chlorophyll per hour and was calculated to be associated with an apparent ATP formation of only 2 micromoles/milligram chlorophyll per hour. Thus, electron flow from H2O to O2 did not result in ATP formation significantly above that produced during NADP reduction.

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Selected References

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  1. Allen J. F. A two-step mechanism for the photosynthetic reduction of oxygen by ferredoxin. Biochem Biophys Res Commun. 1975 Sep 2;66(1):36–43. doi: 10.1016/s0006-291x(75)80291-9. [DOI] [PubMed] [Google Scholar]
  2. Arnon D. I., Chain R. K. Role of oxygen in ferredoxin-catalyzed cyclic photophosphorylations. FEBS Lett. 1977 Oct 15;82(2):297–302. doi: 10.1016/0014-5793(77)80606-6. [DOI] [PubMed] [Google Scholar]
  3. Ben-Hayyim G., Avron M. Mn2+ as electron donor in isolated chloroplasts. Biochim Biophys Acta. 1970 Apr 7;205(1):86–94. doi: 10.1016/0005-2728(70)90064-2. [DOI] [PubMed] [Google Scholar]
  4. Egneus H., Heber U., Matthiesen U., Kirk M. Reduction of oxygen by the electron transport chain of chloroplasts during assimilation of carbon dioxide. Biochim Biophys Acta. 1975 Dec 11;408(3):252–268. doi: 10.1016/0005-2728(75)90128-0. [DOI] [PubMed] [Google Scholar]
  5. Epel B. L., Neumann J. The mechanism of the oxidation of ascorbate and MN2+ by chloroplasts. The role of the radical superoxide. Biochim Biophys Acta. 1973 Dec 14;325(3):520–529. doi: 10.1016/0005-2728(73)90211-9. [DOI] [PubMed] [Google Scholar]
  6. Heber U. Stoichiometry of reduction and phosphorylation during illumination of intact chloroplasts. Biochim Biophys Acta. 1973 Apr 27;305(1):140–152. doi: 10.1016/0005-2728(73)90239-9. [DOI] [PubMed] [Google Scholar]
  7. Lilley R. M. Isolation of Functionally Intact Rhodoplasts from Griffithsia monilis (Ceramiaceae, Rhodophyta). Plant Physiol. 1981 Jan;67(1):5–8. doi: 10.1104/pp.67.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lumsden J., Hall D. O. Chloroplast manganese and superoxide. Biochem Biophys Res Commun. 1975 May 19;64(2):595–602. doi: 10.1016/0006-291x(75)90363-0. [DOI] [PubMed] [Google Scholar]
  9. MEHLER A. H. Studies on reactions of illuminated chloroplasts. I. Mechanism of the reduction of oxygen and other Hill reagents. Arch Biochem Biophys. 1951 Aug;33(1):65–77. doi: 10.1016/0003-9861(51)90082-3. [DOI] [PubMed] [Google Scholar]
  10. Marsho T. V., Behrens P. W. Photosynthetic oxygen reduction in isolated intact chloroplasts and cells in spinach. Plant Physiol. 1979 Oct;64(4):656–659. doi: 10.1104/pp.64.4.656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Nelson N., Neumann J. Interaction between ferredoxin and ferredoxin nicotinamide adenine dinucleotide phosphate reductase in pyridine nucleotide photoreduction and some partial reactions. I. Inhibition of ferredoxin nicotinamide adenine dinucleotide phosphate reductase by ferredoxin. J Biol Chem. 1969 Apr 10;244(7):1926–1931. [PubMed] [Google Scholar]
  12. Patterson C. O., Myers J. Photosynthetic Production of Hydrogen Peroxide by Anacystis nidulans. Plant Physiol. 1973 Jan;51(1):104–109. doi: 10.1104/pp.51.1.104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Radmer R., Kok B., Ollinger O. Kinetics and Apparent K(m) of Oxygen Cycle under Conditions of Limiting Carbon Dioxide Fixation. Plant Physiol. 1978 Jun;61(6):915–917. doi: 10.1104/pp.61.6.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Radmer R., Ollinger O. Light-driven Uptake of Oxygen, Carbon Dioxide, and Bicarbonate by the Green Alga Scenedesmus. Plant Physiol. 1980 Apr;65(4):723–729. doi: 10.1104/pp.65.4.723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Robinson J. M., Smith M. G., Gibbs M. Influence of Hydrogen Peroxide upon Carbon Dioxide Photoassimilation in the Spinach Chloroplast: I. HYDROGEN PEROXIDE GENERATED BY BROKEN CHLOROPLASTS IN AN "INTACT" CHLOROPLAST PREPARATION IS A CAUSAL AGENT OF THE WARBURG EFFECT. Plant Physiol. 1980 Apr;65(4):755–759. doi: 10.1104/pp.65.4.755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. SHIN M., TAGAWA K., ARNON D. I. CRYSTALLIZATION OF FERREDOXIN-TPN REDUCTASE AND ITS ROLE IN THE PHOTOSYNTHETIC APPARATUS OF CHLOROPLASTS. Biochem Z. 1963;338:84–96. [PubMed] [Google Scholar]
  17. Steiger H. M., Beck E. Oxygen Concentration in Isolated Chloroplasts during Photosynthesis. Plant Physiol. 1977 Dec;60(6):903–906. doi: 10.1104/pp.60.6.903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Walker D. A., Ludwig L. J., Whitehouse D. G. Oxygen evolution in the dark following illumination of chloroplasts in the presence of added manganese. FEBS Lett. 1970 Feb 16;6(3):281–284. doi: 10.1016/0014-5793(70)80078-3. [DOI] [PubMed] [Google Scholar]
  19. Ziem-Hanck U., Heber U. Oxygen requirement of photosynthetic CO2 assimilation. Biochim Biophys Acta. 1980 Jul 8;591(2):266–274. doi: 10.1016/0005-2728(80)90158-9. [DOI] [PubMed] [Google Scholar]
  20. de la Roche A. I. Increase in linolenic Acid is not a prerequisite for development of freezing tolerance in wheat. Plant Physiol. 1979 Jan;63(1):5–8. doi: 10.1104/pp.63.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]

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