The kinetics and quantum yield of photophosphorylation in Anacystis nidulans (Richt.) Drouet

Abstract

The active metabolite, ATP, serves not only as a high energy intermediate but also as a controller of some enzymatic reactions. In plant cells, the larger part of the ATP is formed by photophosphorylation. In this paper the rates, the quantum yields, and the wavelength dependencies of photophosphorylation in the blue-green alga Anacystis nidulans are reported. A fluorometric method for determination of enzymatically produced NADPH from ATP was adapted for use on cell extracts. In the light, the ATP level was 0.15 to 0.25 µmoles/mg chl. In the dark, the ATP level was 70% of that in light. In both darkness and anaerobosis, the level was 20%. A return to the light restored the ATP level from both conditions. Dark, anaerobic cells were exposed to measured irradiancies of 710 nm and 620 nm. The rate of ATP formation was measured within the first few seconds and found to be directly proportional to absorbed intensity. Saturation of the rates occurred at an intensity one-tenth the optimum for oxygen production. Quantum requirements of 6-8 were similar for each of the two wavelengths. The system II inhibitor DCMU, had a greater effect at 620 nm that at 710 nm indicating an involvement of system II in photophosphorylation only at 620 nm. At low intensities and over long time periods white light failed to produce a saturating steady-state level of ATP indicating a simultaneous consumption of ATP. Measurements in short dark periods following marginal illumination showed consumption of ATP to be 2 to 4 times greater that production in weak light. Thus, the quantum requirement can be calculated to be 2. ATP formation, therefore, is not the limit ing factor in co2 fixation. The evidence is the high quantum yield of photophosphorylation and the unsaturation of co2 fixation at intensities at which ATP synthesis is saturated.