Carbonyl sulfide (COS), a substrate for carbonic anhydrase, inhibited alkalization of the medium, 02 evolution, dissolved inorganic carbon accumulation, and photosynthetic CO2 fixation at pH 7 or higher by five species of unicellular green algae that had been air-adapted for forming a C02-concentrating process. This COS inhibition can be attributed to inhibition of extenal HC03-conversion to CO2 and OH-by the carbonic anhydrase component of an active CO2 pump. At a low pH of 5 to 6, COS stimulated 02 evolution during photosynthesis by algae with low CO2 in the media without alkalization of the media. This is attributed to some COS hydrolysis by carbonic anhydrase to CO2. Although COS had less effect on HC03-accumulation at pH 9 by a HCO3-pump in Scenedesmus, COS reduced 02 evolution probably by inhibiting internal carbonic anhydrases. Because COS is hydrolyzed to CO2 and H2S, its inhibition of the CO2 pump activity and photosynthesis is not accurate, when measured by 02 evolution, by NaH'4CO3 accumulation, or by 14CO2 fixation. Unicellular green algae, Chlamydomonas, (3, 18), Dunaliella (12), Chlorella (17), Scenesdesmus (1 1, 22), and species of Cyanobacteria (1), when grown or adapted at air levels of C02, form a DIC3-concentrating mechanism, which is suppressed when grown with elevated levels of CO2 (1-5% CO2 in air v/v). Depending on the pH of the medium and algal species, both HCO3-and/or CO2 may enter the cells. Many unicellular green algae have only a CO2 pump, but Scenedesmus has both a CO2 pump and HC03-pump (22