Hydrogen is a renewable fuel that can be biologically produced by green algae in closed anaerobic photobioreactors with light and organic carbon as energy sources. This research aimed to investigate the influence of sulfur concentration and light intensity on hydrogen and ethanol production, as well as on nutrient removal by Chlamydomonas reinhardtii (CC425) in batch cultures. The strain was cultivated in two phases: in the first step, the cultures were maintained under aerobic conditions to obtain biomass; in the second step, the biomass was transferred to closed anaerobic photobioreactors for gas generation under continuous illumination. A factorial design was accomplished to improve the biomass production in the first step, with light variation, pH, and glucose addition. Afterward, light intensity and sulfur concentration were varied to test hydrogen production in the second step. The best hydrogen production occurred in photobioreactors without sulfur addition (average increase of 7 times in the production) and under higher light intensity the productivity was 37% higher than lower light intensity (39.64 ± 2.44 µmol H₂ L⁻¹ h⁻¹). There was an effect of sulfur concentration in the ethanol production and under higher light intensity the production was higher (203.20 ± 31.49 mg L⁻¹). Furthermore, under conditions with the presence of sulfur, there was greater removal of ammoniacal nitrogen (5.3%), phosphate (85.0%), COD (9.1%) and acetic acid (97.2%). This research demonstrates the efficient production of hydrogen by C. reinhardtii and it shows that the process can be associated with ethanol production and nutrient removal.

氢是一种可再生燃料，可以由绿藻在封闭的厌氧光生物反应器中以光和有机碳作为能源进行生物生产。本研究旨在研究硫浓度和光照强度对氢气和乙醇生产以及莱茵衣藻去除养分的影响(CC425) 分批培养。该菌株分两个阶段培养：第一步，将培养物保持在有氧条件下以获得生物量；第二步，将生物质转移到封闭的厌氧光生物反应器中，在连续光照下产生气体。在第一步中，通过光照变化、pH 值和葡萄糖添加，完成了因子设计以提高生物质产量。然后，改变光照强度和硫浓度以测试第二步中的氢气产量。最好的产氢发生在不添加硫的光生物反应器中（产量平均增加 7 倍），在较高光强度下，生产率比较低光强度（39.64 ± 2.44 µmol H ₂ L ^-1  h）高 37%^-1 )。乙醇生产中存在硫浓度的影响，并且在较高的光强度下生产较高（203.20 ± 31.49 mg L ^-1）。此外，在硫存在的条件下，氨氮 (5.3%)、磷酸盐 (85.0%)、COD (9.1%) 和乙酸 (97.2%) 的去除率更高。该研究证明了莱茵衣藻高效生产氢气，并表明该过程可与乙醇生产和营养物去除有关。