Carbonic anhydrase isoforms of Neopyropia yezoensis: Intracellular localization and expression profiles in response to inorganic carbon concentration and life stage.

Macroalgae, particularly commercially grown seaweed, substantially contribute to CO₂ removal and carbon storage. However, knowledge regarding the CO₂ concentrating mechanism (CCM) of macroalgae is limited. Carbonic anhydrase (CA), a key component of the biophysical CCM, plays important roles in many physiological reactions in various organisms. Few characteristics of CA in Neopyropia yezoensis are known, particularly its intracellular location and responses to different concentrations of Ci. We identified, amplified, and characterized 11 putative genes encoding N. yezoensis CA. The predicted corresponding proteins clustered into three subfamilies: α-, β-, and γ-type. The intracellular localization of seven CA isoforms-one in the chloroplasts, three in the cytoplasm, and three in the mitochondria-was elucidated with fusion proteins. Higher NyCA expression, particularly of certain chloroplastic, cytosolic, and mitochondrial CAs, is observed more often during the foliose stage, thus suggesting that CAs play important roles in development in N. yezoensis.