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Planta

, Volume 159, Issue 1, pp 77–83 | Cite as

Oxygen evolution by a reconstituted spinach chloroplast system in the presence ofl-glutamine and 2-oxoglutarate

  • J. W. Anderson
  • D. A. Walker
Article

Abstract

Intact chloroplasts prepared from summer-grown spinach plants supported (aspartate plus 2-oxoglutarate)-dependent O2 evolution but not (glutamine plus 2-oxoglutarate)-dependent O2 evolution. The former activity, which was sensitive to amino oxyacetate, was attributed to transaminase activity and reduction of the resulting oxalo-acetate to malate using H2O as eventual electron donor. A reconstituted chloroplast system which included chloroplast stroma, thylakoid membranes, ferredoxin and NADP(H) supported O2 evolution in the presence ofl-glutamine and 2-oxoglutarate at rates of 15–22 μmol mg-1 chlorophyll h-1 although lower rates were obtained with material from winter-grown plants. Activity was not observed in the absence of ferredoxin and omission of NADP(H) decreased activity by 40%. The reaction was associated with the production of 0.49 mol O2 mol-1 2-oxoglutarate consumed and up to 0.46 mol O2 mol-1 glutamine supplied. The reaction, which was inhibited by azaserine but not by methionine sulphoximine or amino oxyacetate, was attributed to light-coupled glutamate synthase (EC 1.4.1.13) with H2O serving as eventual electron donor. Activity was not affected significantly byl-malate. The reconstituted system also supported O2 evolution in the presence of nitrite, oxaloacetate, (aspartate plus 2-oxoglutarate) and oxidised glutathione.

Key words

Chloroplast (O2 evolution) Glutamate synthase Glutamine metabolism 2-Oxoglutarate metabolism Oxygen evolution (chloroplast) Spinacia (glutamate synthase) 

Abbreviations

Chl

chlorophyll

GSSG

oxidised glutathione

Hepes

4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid

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Copyright information

© Springer-Verlag 1983

Authors and Affiliations

  • J. W. Anderson
    • 1
  • D. A. Walker
    • 1
  1. 1.A.R.C. Research Group on Photosynthesis, Botany DepartmentUniversity of SheffieldSheffieldUK

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