Carbonic anhydrase (CA) is a central enzyme to both transport and metabolic processes at the cellular level. In metabolically active tissue such as muscle, CA in the cytoplasm and on the sarcolemma appears to be important in facilitating CO2 transport out of the cell. Membrane-associated CA, with an extracellular orientation, also appears to be important in acidifying the outer boundary layer through the catalyzed hydration of excreted CO2. This facilitates cellular ammonia transport by providing H+ ions for the protonation of NH3, thus maintaining the trans-membrane NH3 gradient. Mitochondrial CA is known to supply HCO3- for the initial reactions of gluconeogenesis and ureagenesis in mammalian tissues, but systematic comparative studies of CA as a metabolic enzyme are lacking. CA probably evolved as an enzyme of trans-membrane facilitated CO2 transport and took on a secondary metabolic role later in metazoan evolution.