A Stepwise NaHSO₃ Addition Mode Greatly Improves H₂ Photoproduction in Chlamydomonas reinhardtii

NaHSO₃ addition greatly increases the yield of H₂ photoproduction in a unicellular green alga Chlamydomonas reinhardtii through removing O₂ and activating hydrogenase but significantly impairs the activity of PSII, an electron source for H₂ photoproduction. Here, a stepwise addition mode of total 13 mM NaHSO₃, an optimal concentration for H₂ photoproduction of C. reinhardtii identified in a previous one step addition method, significantly improved H₂ photoproduction. Such improvement was believed to be the result of increased residual PSII activity in an anaerobic background, but was at least independent of two alternative electron sinks for H₂ photoproduction, cyclic electron transport around PSI and CO₂ assimilation. Based on the above results, we propose that increased residual PSII activity in an anaerobic environment is an efficient strategy to enhance H₂ photoproduction in C. reinhardtii, and the stepwise NaHSO₃ addition mode is a case study in the strategy.