Biochar feedstock and production method affects its physicochemical properties and subsequent application. This study investigated the effects of biochar from switchgrass (SGB) and poultry litter (PLB) produced at 350 and 700 °C on alcohol formation using CO:CO2:H2 (40:30:30) by Clostridium carboxidivorans (P7) and C. ragsdalei (P11). Fermentations were performed in 250 mL bottles with a 50 mL working volume at 37 °C. Strains P7 and P11 produced 1.2- to 2.2-fold more alcohol and consumed 1.2- to 1.9-fold more syngas using biochars made at 700 °C compared to 350 °C. Both strains also produced 1.4- to1.9-fold more alcohol with both biochars made at 700 °C compared to control without biochar. Strain P11 produced 1.1- and 1.6-fold more alcohol and fatty acids, respectively, in medium with PLB made at 700 °C compared to strain P7. These results provide guidance towards the selection of biochar type and production temperature to improve syngas fermentation.