Level of photosynthetic intermediates in isolated spinach chloroplasts.

The level of intermediates of the photosynthetic carbon cycle was measured in intact spinach chloroplasts in an attempt to determine the cause of the induction lag in CO(2) assimilation. In addition, transient changes in the level of the intermediates were determined as affected by a light-dark period and by the addition of an excess amount of bicarbonate during a period of steady photosynthesis. Assayed enzymically were: ribulose 1,5-diphosphate, pentose monophosphates (mixture of ribose 5-phosphate, ribulose 5-phosphate and xylulose 5-phosphate, hexose monophosphates (mixture of glucose 6-phosphate, glucose 1-phosphate, and fructose 6-phosphate), glyceraldehyde 3-phosphate, dihydroxyacetone phosphate, glycerate acid 3-phosphate, a mixture of fructose 1,6-diphosphate and sedoheptulose 1,7-diphosphate, adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP).The lag in CO(2) fixation appeared to be the result of low levels of pentose monophosphates. The level of ribulose 1,5-diphosphate was roughly equal in chloroplasts showing immediate linear kinetics with respect to CO(2) fixation and chloroplasts which exhibited an initial lag.Following a light-dark transition, CO(2) fixation ceased immediately but the level of glycerate 3-phosphate increased while ribulose 1,5-diphosphate was only slightly effected. The increase in level of glycerate 3-phosphate was correlated with a decrease in triose phosphate. Within 3 to 5 min in the light, ATP reached a maximum concentration while in darkness, all was utilized in 30 to 60 sec. The rapid loss of ATP was ascribed to an ATPase rather than to its utilization in kinase reactions.A rapid increase in CO(2) concentration enhanced the level of triose phosphate, but the level of glycerate 3-phosphate showed only a small overshoot and was considered as evidence that reducing power was not a rate limiting factor. Data were obtained indicating that triose phosphates similar to pentose monophosphates and in contrast to fructose 6-phosphate, glucose 6-phosphate and glucose 1-phosphate could be transported between chloroplast and suspending medium. Differential import and export of phosphorylated compounds may serve as routes alternative to starch and sucrose for the flow of carbon into biosynthetic pathways.