Department of Pharmaceutical Engineering, Woosuk University, Wanju, Korea.
Mol Microbiol. 2010 Mar;75(6):1372-88. doi: 10.1111/j.1365-2958.2010.07058.x. Epub 2010 Feb 8.
Carbon dioxide (CO(2)) and its hydration product bicarbonate (HCO(3)(-)) are essential molecules in various physiological processes of all living organisms. The reversible interconversion between CO(2) and HCO(3)(-) is in equilibrium. This reaction is slow without catalyst, but can be rapidly facilitated by Zn(2+)-metalloenzymes named carbonic anhydrases (CAs). To gain an insight into the function of multiple clades of fungal CA, we chose to investigate the filamentous fungi Aspergillus fumigatus and A. nidulans. We identified four and two CAs in A. fumigatus and A. nidulans, respectively, named cafA-D and canA-B. The cafA and cafB genes are constitutively, strongly expressed whereas cafC and cafD genes are weakly expressed but CO(2)-inducible. Heterologous expression of the A. fumigatus cafB, and A. nidulans canA and canB genes completely rescued the high CO(2)-requiring phenotype of a Saccharomyces cerevisiae Deltance103 mutant. Only the DeltacafA DeltacafB and DeltacanB deletion mutants were unable to grow at 0.033% CO(2), of which growth defects can be restored by high CO(2). Defects in the CAs can affect Aspergilli conidiation. Furthermore, A. fumigatus DeltacafA, DeltacafB, DeltacafC, DeltacafD and DeltacafA DeltacafB mutant strains are fully virulent in a low-dose murine infection.
二氧化碳(CO(2))及其水合产物碳酸氢盐(HCO(3)(-))是所有生物体各种生理过程中必不可少的分子。CO(2)和 HCO(3)(-)之间的可逆相互转化处于平衡状态。没有催化剂时,这种反应很慢,但可以被命名为碳酸酐酶(CA)的 Zn(2+)-金属酶迅速促进。为了深入了解真菌 CA 的多个进化枝的功能,我们选择研究丝状真菌烟曲霉和构巢曲霉。我们分别在烟曲霉和构巢曲霉中鉴定出 4 种和 2 种 CA,分别命名为 cafA-D 和 canA-B。cafA 和 cafB 基因是组成型的,强烈表达,而 cafC 和 cafD 基因表达较弱,但可被 CO(2)诱导。在酿酒酵母 Deltance103 突变体中异源表达烟曲霉 cafB 和构巢曲霉 canA 和 canB 基因完全挽救了高 CO(2)需求表型。只有 DeltacafA DeltacafB 和 DeltacanB 缺失突变体在 0.033% CO(2)下无法生长,其中生长缺陷可通过高 CO(2)恢复。CA 的缺陷会影响曲霉菌的分生孢子形成。此外,烟曲霉 DeltacafA、DeltacafB、DeltacafC、DeltacafD 和 DeltacafA DeltacafB 突变株在低剂量小鼠感染中完全具有毒力。