Hydrogen production was studied using immobilized green alga Chlorella sp. through a two-stage cyclic process where immobilized cells were first incubated in oxygenic photosynthesis followed by anaerobic incubation for H2 production in the absence of sulfur. Chlorella sp. used in this study was capable of generating H2 under immobilized state in agar. The externally added glucose enhanced H2 production rates and total produced volume while shortened the lag time required for cell adaptation prior to H2 evolution. The rate of hydrogen evolution was increased as temperature increased, and the maximum evolution rate under 30 mM glucose was 183 mL/h/L and 238 mL/h/L at 37 °C and 40 °C, respectively. In order to continue repeated cycles of H2 production, at least two days of photosynthesis stage should be allowed for cells to recover H2 production potential and cell viability before returning to H2 production stage again.
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