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In the present work, it was attempted to produce H₂ from food waste by the two-stage fermentation system. Food waste was acidified to lactate by using indigenous lactic acid bacteria under mesophilic condition, and the lactate fermentation effluent (LFE) was subsequently converted to H₂ by photo-fermentation. Rhodobacter sphaeroides KD131 was used as the photo-fermenting bacteria. The optimal conditions for lactate fermentation were found to be pH of 5.5 and substrate concentration of 30 g Carbo. COD/L, under which yielded 1.6 mol lactate/mol glucose. By filtering the LFE and adding trace metal, H₂ production increased by more than three times compared to using raw LFE, and finally reached the H₂ yield of 3.6 mol H₂/mol lactate. Via the developed two-stage fermentation system H₂ yield of 5.8 mol H₂/mol glucose was achieved from food waste, whose value was the highest that ever recorded.

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