Growth of the obligately chemolithotrophic Thiomicrospira sp. strain L-12, isolated from a hydrothermal vent at a depth of 2,550 m in the Galapagos Rift region, was optimal at pH 8 and required 200 mM Na+ and divalent ions (Ca2+ and Mg2+). The organism was microaerophilic and tolerated 300 microM sulfide without a decrease in the rate of CO2 incorporation. Growth and CO2 incorporation occurred within the temperature range of 10 to 35 degrees C, with both optimal at 25 degrees C. At the in situ pressure of 250 atm. the rate of CO2 incorporation was reduced by 25% relative to that measured at 1 atm: it was entirely suppressed at 500 atm. The results of this physiological characterization suggest that Thiomicrospira sp. strain L-12 can be an active autotroph in the hydrothermal environment.