The production of biophotolytic hydrogen (H2) relies on the effective management of oxygen (O2) levels. Coculturing bacteria with microalgae helps mitigate the excess O2 produced by algal cells. After depleting O2, the bacteria activate the enzyme hydrogenase in microalgae, leading to H2 production. In this study, Chlamydomonas reinhardtii was cocultured with indigenous bacteria from activated sludge at varying algae-to-bacteria ratios (1:1, 1:1.5, 1:2, 1:2.5, and 1:3 v/v), with an illumination intensity of 2.8 mmol/m2/s (31 × 103 lux). The 1:1.5 v/v ratio yielded the highest H2 volume (1162 mL/L) and the highest O2 concentration (153.2 mL/L) over a 6-day period. Production of all gaseous components ceased for all ratios as the pH dropped below 4 due to acetate accumulation, and the concentration of acetate reached approximately 1 g/L by the end of each experiment. Gas composition analysis after the first day of coculture revealed that H2, CO2, N2, and O2 constituted 25%–46%, 20%–40%, 5%–30%, and 1%–10% of the total gas volume, respectively. Glucose (10 g/L) was introduced as an external carbon source for all cultures. After 6 days, the coculture maintained a high total organic carbon (TOC) level of 3.1 g/L, whereas the initial TOC ranged between 3.9 and 4.3 g/L. The findings illustrated a significant correlation between H2 production, acetate accumulation levels, and O2 consumption. The algae–activated sludge coculture method substantially enhanced H2 production compared with previously published methods employing only one or two types of bacterial cultures, underscoring its potential for more efficient biophotolytic H2 production.
Read full abstract