A spontaneous mutant of the cyanobacterium Synechocystis PCC6803 was isolated for its resistance to acetazolamide, an inhibitor of carbonic anhydrase. The mutant showed a deficiency in oxygen exchange between CO(2) and H(2)O, a lower level of stable internal CO(2) pool and a decreased capacity to adapt its photosynthetic affinity under limited inorganic carbon regime. The initial rate of uptake of inorganic carbon was identical to that of wild-type cells. It is demonstrated that the mutation affects the carbonic anhydrase activity. This could result from either of two impairments: a deficiency in the enzyme activity detectable by mass spectrometric determinations, or a modification of the cellular compartment in which the enzyme is located, preventing its activity.