Oxygenic photosynthetic organisms perform solar energy conversion of water and CO₂ to O₂ and sugar at a broad range of wavelengths and light intensities. These cells also metabolize sugars using a respiratory system that functionally overlaps the photosynthetic apparatus. In this study, we describe the harvesting of photocurrent used for hydrogen production from live cyanobacteria. A non-harmful gentle physical treatment of the cyanobacterial cells enables light-driven electron transfer by an endogenous mediator to a graphite electrode in a bio-photoelectrochemical cell, without the addition of sacrificial electron donors or acceptors. We show that the photocurrent is derived from photosystem I and that the electrons originate from carbohydrates digested by the respiratory system. Finally, the current is utilized for hydrogen evolution on the cathode at a bias of 0.65 V. Taken together, we present a bio-photoelectrochemical system where live cyanobacteria produce stable photocurrent that can generate hydrogen.

Significance: Protection from overexcitation is one of the most important requirements of all photosynthetic organisms. Here we present a model based on coupled cross-linking/mass spectrometry and site-directed mutagenesis of the means by which the orange carotenoid protein (OCP) binds to the phycobilisome (PBS) antenna complex to avoid photodamage. The model shows that the protein must actively burrow into the complex, separating the PBS rings in the process. This penetration explains for the first time, to our knowledge, how the OCP carotenoid could approach the PBS chromophores at a distance that enables nonphotochemical quenching. However, the alteration in the core structure caused by OCP binding could also prevent energy transmission to the reaction centers.

Previous studies have shown that live cyanobacteria can produce photocurrent in bio-photoelectrochemical cells (BPECs) that can be exploited for clean renewable energy production. Electron transfer from cyanobacteria to the electrochemical cell was proposed to be facilitated by small molecule(s) mediator(s) whose identity (or identities) remain unknown. Here, we elucidate the mechanism of electron transfer in the BPEC by identifying the major electron mediator as NADPH in three cyanobacterial species. We show that an increase in the concentration of NADPH secreted into the external cell medium (ECM) is obtained by both illumination and activation of the BPEC. Elimination of NADPH in the ECM abrogates the photocurrent while addition of exogenous NADP+ significantly increases and prolongs the photocurrent production. NADP+ is thus the first non-toxic, water soluble electron mediator that can functionally link photosynthetic cells to an energy conversion system and may serve to improve the performance of future BPECs.