Abstract Microorganisms used in syngas fermentation require nutrients to grow and convert syngas (CO, H2 and CO2) into various products. Many of the essential nutrients can be provided by biochar. Poultry litter biochar (PLBC) contains minerals and trace metals and has a high pH buffering capacity, making it suitable as a nutrient supplement. The effects of PLBC loadings from 1 to 20 g L−1 on syngas fermentation were determined in 250 ml bottle assays. Results showed that 10 and 20 g L−1 PLBC significantly increased ethanol production compared to standard yeast extract (YE) medium. Fermentations in a 3L continuous stirred tank reactor (CSTR) with 10 g L−1 PLBC with and without 4-morpholineethanesulfonic acid (MES) showed 64% and 36% more ethanol production, respectively, than standard medium. The acetic acid accumulated at the beginning of fermentation was completely converted to ethanol in all media tested in the CSTR. These results demonstrate the feasibility of using PLBC medium without costly MES in the CSTR to enhance ethanol production from syngas for potential use at commercial scale.