The filamentous ascomycete Sordaria macrospora can survive in ambient air without carbonic anhydrases

The rapid interconversion of carbon dioxide and bicarbonate (hydrogen carbonate) is catalysed by metalloenzymes termed carbonic anhydrases (CAs). CAs have been identified in all three domains of life and can be divided into five evolutionarily unrelated classes (, , , and) that do not share significant sequence similarities. The function of the mammalian, prokaryotic and plant -CAs has been intensively studied but the function of CAs in filamentous ascomycetes is mostly unknown. The filamentous ascomycete Sordaria macrospora codes for four CAs, three of the -class and one of the -class. Here, we present a functional analysis of CAS4, the S. macrospora -class CA. The CAS4 protein was post-translationally glycosylated and secreted. The knockout strain cas4 had a significantly reduced rate of ascospore germination. To determine the cas genes required for S.macrospora growth under ambient air conditions, we constructed double and triple mutations of the four cas genes in all possible combinations and a quadruple mutant. Vegetative growth rate of the quadruple mutant lacking all cas genes was drastically reduced compared to the wild type and invaded the agar under normal air conditions. Likewise the fruiting bodies were embedded in the agar and completely devoid of mature ascospores.