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emi2859-sup-0001-si.docx1389K

Fig. S1. Effect of nitrite on metabolic activity of D. vulgaris strains in closed serum bottles at high (8–10 mM) sulfide and a headspace of 10% CO2 and 90% N2. Nitrite was added at mid-log phase ([DOWNWARDS ARROW]) to a final concentration of 5 mM. Lactate (▴) and acetate (▾) concentrations (left panels), as well as sulfide (•) and nitrite (▪) concentrations (right panels) were monitored as a function of time (h). Experiments were conducted with at least two replicates and the data are represented with standard error bars.

emi2859-sup-0001-si.docx1389K

Fig. S2. Effect of nitrite on metabolic activity of D. vulgaris strains in closed serum bottles with high (8–10 mM) sulfide in the aqueous phase and H2 (5% vol/vol), CO2 (10%) and N2 in the headspace. Nitrite was added at mid-log phase ([DOWNWARDS ARROW]) to a final concentration of 10 mM. Lactate (▴) and acetate (▾) concentrations (left panels), as well as sulfide (•) and nitrite (▪) concentrations (right panels) were monitored as a function of time (h). Experiments were conducted with at least two replicates and the data are represented with standard error bars.

emi2859-sup-0001-si.docx1389K

Fig. S3. Effect of nitrite on the growth of D. vulgaris Hildenborough strains at low (0–1 mM) sulfide. Growth was in side-arm flasks with WP-LS medium in an anaerobic hood with an atmosphere of 5% H2, 10% CO2 and 85% N2 under conditions of gas exchange. Nitrite (10 mM) was added at mid-log phase ([DOWNWARDS ARROW]). Results are averages for triplicate determinations with standard error bars as indicated; 149 Klett units = 1 OD600.

emi2859-sup-0001-si.docx1389K

Fig. S4. Effect of 10 mM nitrite ([DOWNWARDS ARROW]) on metabolic activity of the Hcp2 GEI strain complemented with plasmids containing the roo1 gene, the hcp1 gene, or no insert in closed serum bottles at high (8–10 mM) sulfide and a 10% CO2, 90% N2 headspace. Lactate (▴) and acetate (▾) concentrations (left panels), as well as sulfide (•) and nitrite (▪) concentrations (right panels) were monitored as a function of time (h). Experiments were conducted with at least two replicates and the data are represented with standard error bars.

emi2859-sup-0001-si.docx1389K

Table S1. Primers used throughout this study. The target description as well as sequence is shown. Uppercase sequence refers to the complementary region and lowercase sequence indicates a tag (usually with a restriction site to facilitate cloning).

emi2859-sup-0001-si.docx1389K

Table S2. Vectors and constructs used in this study for gene replacement mutagenesis and mutant complementation.

emi2859-sup-0001-si.docx1389K

Table S3. Wild-type and mutant strains used in this study.

emi2859-sup-0001-si.docx1389K

Table S4. Construction of mutant strains used in this study.

emi2859-sup-0001-si.docx1389K

Table S5. Rates of nitrite reduction of strains used in this study with 10 mM nitrite added at mid-log phase, where cultures had approximately 10 mM sulfide and 20 mM of residual sulfate. The serum bottle headspace was 10% CO2 and 90% N2, except for entries marked (H2), which contained 5% H2, 10% CO2 and 85% N2. Data have been ranked from highest to lowest RS.

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