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Biological processes run by microalgae are prospective but still little known methods of hydrogen production. A prerequisite for their increased advancement is the development of economically viable and efficient technologies. The study presented in this manuscript focused on determining the efficiency of biohydrogen production by <i>P. subcordiformis</i> using a culture medium prepared based on natural waters. The rate of <i>P. subcordiformis</i> biomass growth reached 317.6 ± 42.3 mg_ODM/dm³·d and ensured a biomass concentration of 3493 ± 465 mg_ODM/dm³. The percentage concentration of hydrogen in the biogas reached 63.2 ± 1.4%, and its production rate ranged from 0.53 ± 0.05 cm³/h to 0.70 ± 0.01 cm³/h.