Previously, a new method developed, by which sucrose is produced as a viable alternative for hydrogen formation via anaerobic dark fermentation of salt stressed PAMCOD strain of cyanobacterium Synechococcus elongatus PCC7942. As cells under such conditions accumulate glycogen, together with sucrose, and degrade it in dark fermentation conditions, we here extended our previous method, forcing glycogen breakdown by nitrogen depletion, which resulted to a significantly augmented, up to 10,000-fold, hydrogen evolution. Importantly, the structural integrity of their phycobilisomes was found not to be affected. Moreover, the sustainability of the procedure was demonstrated by investigating the viability of the cyanobacterial biomass following its dark fermentation; it was found that cells were able to proliferate upon their dark fermentation, both in double BG-11 and in nitrogen-enriched BG-11 medium. The present study demonstrates a significant H₂ production and indicates the glucose derived from the endogenous glycogen as the main carbon substrate fermented.