Using stable isotope labeling to study the nitrogen metabolism in Anabaena flos-aquae growth and anatoxin biosynthesis

Freshwater resources are under stress around the world due to rapid urbanization and excessive water consumption. Cyanobacterial blooms have occurred frequently in surface waters, which produced toxic secondary metabolites causing a potential harm to aquatic ecosystems and humans. In this study, the relationship between different types of nitrogen source and the algal growth of Anabaena flos-aquae , which was isolated from Dianchi Lake in southern China, was investigated. Experiments were accomplished by using four types of isotope tracers including 15 N-ammonium chloride, 15 N-sodium nitrate, 15 N-urea, 15 N- l -alanine in culture medium to characterize the biosynthesis of 15 N-anatoxin-a (ATX-A), which is a major algal toxin from A. flos-aquae , through liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results showed that all these four types of nitrogen can be incorporated into algal cells. The ATX-A production with urea as the nitrogen source was much higher than that with the other three types of nitrogen. The 15 N labeling experiments further demonstrated that the uptake of organic nitrogen nutrients was significantly greater than that of inorganic nitrogen . These results provide new evidence and deeper insight to explore the biosynthesis of ATX-A in the specific strain of A. flos-aquae .