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The main goal of this study was to increase the hydrogen production rate improving the culture technique and the photobioreactor performances. Experiments were carried out at a constant culture temperature of 30°C and at an average irradiance of 480 W m−2using a cylindrical photobioreactor (4.0 cm, internal diameter). The culture technique, namely, the semicontinuous regime for growingRhodopseudomonas palustris42OL made it possible to achieve a very high daily hydrogen production rate of 594 ± 61 mL (H2) L−1 d−1. This value, never reported for this strain, corresponds to about 25 mL (H2) L−1 h−1, and it was obtained when the hydraulic retention time (HRT) was of 225 hours. Under the same growth conditions, a very high biomass production rate (496 ± 45 mg (dw) L−1 d−1) was also achieved. Higher or lower HRTs caused a reduction in both the hydrogen and the biomass production rates. The malic-acid removal efficiency (MAre) was always higher than 90%. The maximal hydrogen yield was 3.03 mol H2mol MA−1at the HRT of 360 hours. The highest total energy conversion efficiency was achieved at the HRT of 225 hours.