External and internal carbonic anhydrases in Dunaliella species

Abstract

Four species of Dunaliella and a carbonic anhydrase deficient mutant of Dunaliella tertiolecta (Butcher), HL 25/8, developed a process for concentrating dissolved inorganic carbon when adapted on air or low CO2 for 24 h in the light. However, the external carbonic anhydrase activities in Dunaliella species were nil or low compared with those in Chlamydomonas reinhardtii (Danegard) or some Chlorella species. D. tertiolecta had low and about equal levels of external and internal carbonic anhydrase. D. parva (Lerche) and the D. tertiolecta mutant had negligible external carbonic anhydrase. D. viridis (Teodoresco) and D. salina (Teodoresco), high salt tolerant species, had high activities of intenal carbonic anhydrase but low levels of external carbonic anhydrase. Antiserum prepared against the 37 kDa peptide of extracellular carbonic anhydrase from Chlamydomonas reinhardtii was immunoreactive with a polypeptide of 30 kDa in D. tertiolecta and its salt sensitive external carbonic anhydrase (CA) mutant HL 25/8. External CA activity from D. tertiolecta was stimulated about three-fold, by including 0.5 M NaCl in the assay medium, while internal CA was not significantly affected by NaCl. External CA activities in the other species were insensitive to NaCl, while their internal activities were 90% inhibited by 0.5 M NaCl. Sorbitol only partly replaced NaCl in stimulating the external CA from D. tertiolecta. These experiments were performed with Dunaliella spp. grown in controlled laboratory cultures during September through November 1989.