G. sulfurreducens strain DL-1 and cytochrome-deletion mutants were cultured from frozen stocks, inoculated into and propagated once into liquid growth medium following the study of Coppi et al. (44). The growth medium was supplemented with 10 mM acetate and 40 mM fumarate as the electron donor and acceptor, respectively, and Wolf’s vitamin and mineral supplement [American Type Culture Collection (ATCC)]. The medium (final pH = 6.8) was degassed for 30 min/liter at 80°C in the anaerobic chamber and transferred to glass pressure tubes with butyl stoppers unless otherwise noted. S. oneidensis strain MR-1 and mutants were inoculated from frozen stocks and grown in LB broth aerobically at 30°C, 200-rpm shaking for 16 hours, and then transferred 1:100 and grown anaerobically at 30°C for ca. 10 hours in 20 ml of Shewanella basal medium (10) supplemented with 100 mM HEPES, 0.2% casamino acids, Wolf’s vitamin, and a mineral supplement (ATCC). The anaerobic growth medium (final pH = 7.0) also contained either 10 mM lactate and 60 mM fumarate or 20 mM lactate and 15 mM Fe(III) citrate supplemented with 10 mM fumarate. Kanamycin was also provided at a concentration of 50 μM/ml for the growth of ΔMtr complementary mutants (ΔMtr + MtrABC, ΔMtr + MtrDEF, and ΔMtr + vector). E. coli strains ccm and ccm + CymA/MtrABC were cultured from frozen stocks and grown aerobically overnight in 2xYT medium at 37°C, 250-rpm shaking, and then transferred 1:100 into 25 ml of 2xYT medium and grown with 250-rpm shaking for 16 hours at 30°C. Then, each strain was centrifuged at 6000 rpm for 4 min and resuspended (with an OD600 ~ 0.7) in 20 ml of the anaerobic M1 medium (42) supplemented with 0.2% casamino acids, 40 mM lactate, 15 mM Fe(III) citrate, and 10 mM fumarate and grown for 5 days at 30°C. The growth medium also contains kanamycin (50 μM/ml) and chloramphenicol (30 μM/ml). Anaerobic culturing, growth media, and transfers were conducted in an anaerobic chamber (Coy Laboratory Products) with a H2:CO2:N2 (5:20:75) atmosphere.

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