Ethanol produced from lignocellulosic biomass is an alternative transportation fuel with the potential to lower greenhouse gas emissions and increase energy security. Source-separated organic waste (SSO) from the city of Toronto was investigated for feasibility as a lignocellulosic ethanol feedstock. Clostridium phytofermentans is a mesophilic, cellulolytic and ethanologenic species with potential application for ethanol production from lignocullulosic biomass. C. phytofermentans was considered for biofuels production through experiments on a variety of substrates including soluble sugars and pure celluloses. Results from this study found that C. phytofermentans produced 73% of the theoretical ethanol yield on cellobiose but grew poorly on glucose and xylose. In addition, C. phytofermentans grew marginally on microcrystalline and ball-milled cellulose, but with supplemental enzymes produced 55% of the theoretical ethanol yield.