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Metabolism and Enzymology

Utilization of Inorganic Carbon by Ulva lactuca

Department of Botany, Tel Aviv University, Tel Aviv 69978, Israel

Thalli discs of the marine macroalga Ulva lactuca were given inorganic carbon in the form of HCO₃^–, and the progression of photosynthetic O₂ evolution was followed and compared with predicted O₂ evolution as based on calculated external formation of CO₂ (extracellular carbonic anhydrase was not present in this species) and its carboxylation (according to the K_m(CO₂) of ribulose-1,5-bisphosphate carboxylase/oxygenase), at two different pHs, assuming a photosynthetic quotient of 1. The K_m(inorganic carbon) was some 2.5 times lower at pH 5.6 than at the natural seawater pH of 8.2, whereas V_max was similar under the two conditions, indicating that the unnaturally low pH per se had no adverse effect on U. lactuca's photosynthetic performance. These results, therefore, could be evaluated with regard to differential CO₂ and HCO₃^– utilization. The photosynthetic performance observed at the lower pH largely followed that predicted, with a slight discrepancy probably reflecting a minor diffusion barrier to CO₂ uptake. At pH 8.2, however, dehydration rates were too slow to supply CO₂ for the measured photosynthetic response. Given the absence of external carbonic anhydrase activity, this finding supports the view that HCO₃^– transport provides higher than external concentrations of CO₂ at the ribulose-1,5-bisphosphate carboxylase/oxygenase site. Uptake of HCO₃^– by U. lactuca was further indicated by the effects of potential inhibitors at pH 8.2. The alleged band 3 membrane anion exchange protein inhibitor 4,4'-diisothiocyanostilbene-2,2'disulphonate reduced photosynthetic rates only when HCO₃^– (but not CO₂) could be the extracellular inorganic carbon form taken up. A similar, but less drastic, HCO₃^–-competitive inhibition of photosynthesis was obtained with Kl and KNO₃. It is suggested that, under ambient conditions, HCO₃^– is transported into cells at defined sites either via facilitated diffusion or active uptake, and that such transport is the basis for elevated internal [CO₂] at the site of ribulose-1,5-bisphosphate carboxylase/oxygenase carboxylation.

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