The efficiency of enzymatic and microbial saccharification of cassava waste for ethanol production was investigated and the effective parameters were optimized. The mixture of amylase and amyloglucosidase (AMG) resulted in a significantly higher rate of saccharification (79.6%) than the amylase alone (68.7%). Simultaneous saccharification and fermentation (SSF) yielded 6.2 g L-1 ethanol representing 64.5% of the theoretical yield. Saccharification and fermentation using pure and co-cultures of fungal isolates including Rhizopus stolonifer, Aspergillus terreus, Saccharomyces diastaticus and Zymomonas mobilis revealed that the co-culture system involving S. diastaticus and Z. mobilis was highly suitable for the bio-conversion of cassava waste into ethanol, resulting in 20.4 g L-1 in 36 h (91.3% of the theoretical yield).