As is well known, carbohydrate is the most appropriate organic material for hydrogen ermentation, and its hydrogen yield is significantly larger than that of protein. The fermentation of protein began with hydrogen production followed by hydrogen consumption, which helps overall hydrogen recovery. Both carbohydrate and protein are basic components of organic material, and yet carbohydrate is known to be a better substrate than protein in terms of hydrogen yield during hydrogen fermentation. This study used multiple substrates containing different ratios of glucose and peptone as multiple substrates to investigate the roles played by carbohydrate and protein in hydrogen fermentation. The experimental results demonstrated that suitable ratios of glucose and peptone improved the growth of hydrogen producing bacteria. Additionally, a maximum hydrogen yield of 6.4 mmole-H2/g-COD was obtained from the multiple ubstrate containing 40% peptone and 60% glucose. Most of the produced hydrogen came from fermentation of glucose, not peptone. During hydrogen fermentation, the pH dropped by 1.0 and 1.9 units in 80% and 20% of peptone content in the substrate. Ammonia produced due to peptone degradation neutralized the acids produced from hydrogen fermentation.

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