The double-layer photobioreactor using two types of photosynthetic bacteria, Rhodobacter sphaeroides RV and its reduced-pigment mutant, MTP4, was developed for efficient hydrogen production. The two types of bacteria had different characteristics on light energy, hydrogen production rate and conversion efficiency. MTP4 produced hydrogen more efficiently under high light conditions and RV did so under low light conditions. Illuminated light toward the surface of a photobioreactor quasi-exponentially declines as it penetrates into the reactor. When two types of bacteria were placed using the developed reactor according to this light distribution, the hydrogen production rate reached 3.64 l/m(2)/h at a light intensity of 500 W/m(2) in 24 h and the conversion efficiency of light energy to hydrogen was 2.18%. These values were 33% higher than those of only using RV. The low light in the deep part of the reactor was utilized efficiently, resulting in a higher hydrogen production rate. (C) 2002 Published by Elsevier Science Ltd on behalf of the International Association for Hydrogen Energy.