Autotrophic hydrogen photoproduction by operation of carbon-concentrating mechanism in Chlamydomonas reinhardtii under sulfur deprivation condition

•Two-stage autotrophic H2 production system was improved by using a functional CCM.•CCM-induced cells sustained low K0.5(CO2) levels during autotrophic S deprivation.•CCM induction elevated starch level of C. reinhardtii in autotrophic S deprivation.•CCM-induced cells showed incredible autotrophic H2 photoproduction at 50 μE/m2/s.

Under autotrophic conditions, starch plays an important role in establishing anoxic conditions during PSII-dependent hydrogen (H2) photoproduction in microalgae. This is because starch is the sole organic substrate during respiratory consumption of internal oxygen (O2) from PSII-dependent direct pathway. Herein, we propose a novel approach to further facilitate the internal starch synthesis of Chlamydomonas reinhardtii through the operation of carbon-concentrating mechanism (CCM) along with a two-stage process based on sulfur (S) deprivation, thereby resulting in enhanced anaerobic capacity during PSII-dependent H2 photoproduction. When CCM-induced cells were exposed to high levels of carbon dioxide (CO2) (5%, v/v) with S deprivation, internal levels of starch were significantly elevated by retaining a functional CCM with the boosted photosynthetic activity during 24 h of O2 evolution phase (I) of S deprivation. Consequently, during H2 production phase of S deprivation at irradiance of 50 μE m−2 s−1, the concentrations of starch and H2 in CCM-induced cells were remarkably enhanced by 65.0% and 218.9% compared to that of CCM-uninduced cells, respectively. The treatment of low-CO2-driven CCM induction prior to S deprivation is a cost-effective and energy-efficient strategy that significantly improves the solar-driven H2 production by microalgae; this is particularly realizable in an industrial scale.