AbstractCyanobacteria have been linked with hydrogen cyanide, based on their ability to catabolize it by the nitrogenase enzyme, as a part of nitrogen fixation. Nitrogenase can also use hydrogen cyanide instead of its normal substrate, dinitrogen and convert it to methane and ammonia. In this study, we tested whether cyanobacteria are able, not only to reduce, but also to produce HCN. The production of HCN was examined in 78 cyanobacteria strains from all five principal sections of cyanobacteria, both non-heterocytous and heterocytous, representing a variety of lifestyles and habitats. Twenty-eight (28) strains were found positive for HCN production, with universal representation amongst 22 cyanobacterial planktic and epilithic genera inhabiting freshwater, brackish, marine (including sponges), and terrestrial (including anchialine) habitats. The HCN production could be linked with nitrogen fixation, as all of HCN producing strains are considered capable of fixing nitrogen. Epilithic lifestyle, where cyanobacteria are more vulnerable to a number of grazers and accumulate more glycine, had the largest percentage (75%) of HCN-producing cyanobacteria compared to strains from aquatic ecosystems. Further, we demonstrate the isolation and characterisation of taxa like Geitleria calcarea and Kovacikia muscicola, for which no strain existed and Chlorogloea sp. TAU-MAC 0618 which is, to the best of our knowledge, the first bacterium isolate from anchialine ecosystems. Our results highlight the complexity of cyanobacteria secondary metabolism, as well as the diversity of cyanobacteria in underexplored habitats, providing a missing study material for this type of environments.