Marine calcification is an important component of the global carbon cycle. The mechanism by which some organisms take up inorganic carbon for the production of their shells or skeletons, however, remains only partly known. Although foraminifera are responsible for a large part of the global calcium carbonate production, the process by which they concentrate inorganic carbon is debated. Some evidence suggests that seawater is taken up by vacuolization and participates relatively unaltered in the process of calcification, whereas other results suggest the involvement of transmembrane transport and the activity of enzymes like carbonic anhydrase. Here, we tested whether inorganic-carbon uptake relies on the activity of carbonic anhydrase using incubation experiments with the perforate, large benthic, symbiont-bearing foraminifer Amphistegina lessonii. Calcification rates, determined by the alkalinity anomaly method, showed that inhibition of carbonic anhydrase by acetazolamide (AZ) stopped most of the calcification process. Inhibition of photosynthesis either by 3-(3,4-Dichlorophenyl)-1,1-dimethylurea (DCMU) or by incubating the foraminifera in the dark also decreased calcification rates but to a lesser degree than with AZ. Results from this study show that carbonic anhydrase plays a key role in biomineralization of Amphistegina lessonii and indicates that calcification of those perforate, large benthic foraminifera might, to a certain extent, benefit from the extra dissolved inorganic carbon (DIC), which causes ocean acidification.