Conditions for Vigorous Growth on Sulfide and Reactor-Scale Cultivation Protocols for the Thermophilic Green Sulfur Bacterium Chlorobium tepidum

ABSTRACT We describe a reactor-scale cultivation protocol for the fastest-growing and only known thermophilic member of the family Chlorobiaceae, Chlorobium tepidum. We discovered that C. tepidum would grow with sulfide as the sole electron source at rates and with final cell yields comparable to those found with thiosulfate only if the sulfide concentration was maintained below 0.1 mM and the culture redox potential was at −300 ± 20 mV. Such was also the requirement for growth in a photobioreactor when thiosulfate (optimum level, 12 mM) was used as the preferred electron source. For cultivation of C. tepidum on a 5- to 500-ml scale, we used the system of Balch and Wolfe (Appl. Environ. Microbiol. 32:781–791, 1976) using stopper-sealed serum tubes and bottles as an alternative to the methods commonly used for the cultivation of phototrophic anaerobes and obtained consistent results.