Enhanced hydrogen production of Rhodobacter sphaeroides promoted by extracellular H+ of Halobacterium salinarum

Purpose This study utilized the principle that the bacteriorhodopsin (BR) produced by Halobacterium salinarum could increase the hydrogen production of Rhodobacter sphaeroides. H. salinarum are co-cultured with R. sphaeroides to determine the impact of purple membrane fragments (PM) on R. sphaeroides and improve its hydrogen production capacity. Methods In this study, low-salinity in 14 % NaCl domesticates H salinarum. Then, 0–160 nmol of different concentration gradient groups of bacteriorhodopsin (BR) and R. sphaeroides was co-cultivated, and the hydrogen production and pH are measured; then, R. sphaeroides and immobilized BR of different concentrations are used to produce hydrogen to detect the amount of hydrogen. Two-chamber microbial hydrogen production system with proton exchange membrane-assisted proton flow was established, and the system was operated. As additional electricity added under 0.3 V, the hydrogen production rate increased with voltages in the coupled system. Results H salinarum can still grow well after low salt in 14% NaCl domestication. When the BR concentration is 80 nmol, the highest hydrogen production reached 217 mL per hour. Both immobilized PC (packed cells) and immobilized PM (purple membrane) of H. salinarum could promote hydrogen production of R. sphaeroides to some extent. The highest production of hydrogen was obtained by the coupled system with 40 nmol BR of immobilized PC, which increased from 127 to 232 mL, and the maximum H2 production rate was 18.2 mL−1 h−1 L culture. In the 192 h experiment time, when the potential is 0.3 V, the hydrogen production amount can reach 920 mL, which is 50.3% higher than the control group. Conclusions The stability of the system greatly improved after PC was immobilized, and the time for hydrogen production of R. sphaeroides significantly extended on same condition. As additional electricity added under 0.3 V, the hydrogen production rate increased with voltages in the coupled system. These results are helpful to build a hydrogen production-coupled system by nitrogenase of R. sphaeroides and proton pump of H. salinarum. Graphical abstract