Evidence for bicarbonate accumulation by Anacystis nidulans

Kinetic studies of photosynthetic O2 evolution as a function of pH were conducted to investigate the nature of the inorganic carbon used during photosynthesis by Anacystis nidulans. At pH 5, the apparent affinity for carbon during photosynthesis was similar in air-grown and high CO2 grown cells, but at alkaline pH, the apparent affinity was much greater in air-grown cells. The substrate concentration for half-maximum rates of photosynthesis in air-grown cells remained constant as a function of pH when the substrate was expressed as total carbon, suggesting that these cells were capable of using varying proportions of CO2 and [Formula: see text]. Photosynthesis in high CO2 grown algae appeared to be more dependent on CO2 over the pH range, indicating that CO2 was the predominant carbon species used, but [Formula: see text] uptake was also indicated. Internal inorganic carbon and photosynthetic carbon fixation in air-grown cells were determined at pH 8.5, using silicone oil centrifugation. Anacystis accumulated inorganic carbon in large excess of that in the external medium by a mechanism which is sensitive to inhibitors of energy metabolism and independent of concurrent carbon fixation; light was required to accumulate and maintain the internal carbon pool. The degree of accumulation was a function of the carbon concentration in the external medium; at 12 μM external carbon, the accumulation ratio was in excess of 100-fold, whereas at 4.76 mM, the ratio was only 5-fold. The rates of carbon transport were always sufficient to maintain photosynthesis. Carbon efflux rates approaching 40% of the influx rate were found at equilibrium internal carbon concentrations. Kinetic parameters of photosynthesis are discussed with reference to the known properties of algal ribulose bisphosphate (RuBP) carboxylase–oxygenase. It is concluded that the internal inorganic carbon pool serves as an intermediate for photosynthetic carbon fixation and that, if CO2 and [Formula: see text] are in equilibrium, the carbon accumulation at ambient CO2 and O2 is sufficient to suppress RuBP oxygenase activity.