<P><B>Abstract</B></P> <P>This study conducted batch experiments to evaluate the potential of butanol production from microalgae biodiesel residues by <I>Clostridium acetobutylicum</I>. The results indicated that with 90g/L of glucose as the sole substrate the highest butanol yield of 0.2g/g-glucose was found, but the addition of butyrate significantly enhanced the butanol yield. The highest butanol yield of 0.4g/g-glucose was found with 60g/L of glucose and 18g/L of butyrate. Using microalgae biodiesel residues as substrate, <I>C. acetobutylicum</I> produced 3.86g/L of butanol and achieved butanol yield of 0.13g/g-carbohydrate via ABE fermentation, but the results indicated that approximately one third of carbohydrate was not utilized by <I>C. acetobutylicum</I>. Biological butanol production from microalgae biodiesel residues can be possible, but further research on fermentation strategies are required to improve production yield.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Butanol production from microalgae biodiesel residues via ABE fermentation. </LI> <LI> More than twofold of butanol yield increase with additional butyrate of 18–20g/L. </LI> <LI> The highest butanol production of 3.86g/L was achieved with residues of 300g/L. </LI> <LI> The highest butanol yield of 0.13g/g-carbohydrate was with residues of 100g/L. </LI> <LI> One third of carbohydrate from residues cannot be utilized by <I>C. acetobutylicum.</I> </LI> </UL> </P>