Photoproduction of H-2 by green algae utilizes electrons originating f
rom the photosynthetic oxidation of water and does not require metabol
ic intermediates. However, algal hydrogenases are extremely sensitive
to O-2, which limits their usefulness in future commercial H-2-product
ion systems. We designed an experimental technique for the selection o
f O-2-tolerant, H-2-producing variants of Chlamydomonas reinhardtii ba
sed on the ability of wild-type cells to survive a short (20 min) expo
sure to metronidazole in the presence of controlled concentrations of
O-2. The number of survivors depends on the metronidazole concentratio
n, light intensity, preinduction of the hydrogenase, and the presence
or absence of O-2. Finally, we demonstrate that some of the selected s
urvivors in fact exhibit H-2-production capacity that is less sensitiv
e to O-2 than the original wild-type population.