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Plant Physiol, January 2000, Vol. 122, pp. 127-136
Sustained Photobiological Hydrogen Gas Production upon Reversible
Inactivation of Oxygen Evolution in the Green Alga
Chlamydomonas reinhardtii1
Anastasios
Melis,*
Liping
Zhang,
Marc
Forestier,
Maria L.
Ghirardi, and
Michael
Seibert
Department of Plant and Microbial Biology, University of
California, 111 Koshland Hall, Berkeley, California 94720-3102 (A.M.,
L.Z.); and Basic Sciences Center, National Renewable Energy
Laboratory, Golden, Colorado 80401-3393 (M.F., M.L.G., M.S.).
The work describes a novel approach
for sustained photobiological production of H2 gas via
the reversible hydrogenase pathway in the
green alga Chlamydomonas reinhardtii. This
single-organism, two-stage H2 production method circumvents
the severe O2 sensitivity of the reversible hydrogenase by
temporally separating photosynthetic O2 evolution and
carbon accumulation (stage 1) from the consumption of cellular
metabolites and concomitant H2 production (stage 2). A
transition from stage 1 to stage 2 was effected upon S deprivation of
the culture, which reversibly inactivated photosystem II (PSII) and
O2 evolution. Under these conditions, oxidative respiration by the cells in the light depleted O2 and caused
anaerobiosis in the culture, which was necessary and sufficient for the
induction of the reversible hydrogenase. Subsequently,
sustained cellular H2 gas production was observed in the
light but not in the dark. The mechanism of H2 production
entailed protein consumption and electron transport from endogenous
substrate to the cytochrome b6-f and PSI complexes in the
chloroplast thylakoids. Light absorption by PSI was required for
H2 evolution, suggesting that photoreduction of ferredoxin
is followed by electron donation to the reversible hydrogenase. The
latter catalyzes the reduction of protons to molecular H2
in the chloroplast stroma.
1
The work was supported by the U.S. Department of
Energy Hydrogen Research and Development Program under Department of
Energy-University of California, Berkeley, Cooperative Agreement (no.
DE-FC36-98GO10278 to A.M. and contract no. DE-AC36-98-GO10337 to
M.L.G. and M.S.).
*
Corresponding author; e-mail melis{at}nature.berkeley.edu; fax
510-642-4995.
© 2000 American Society of Plant Physiologists
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