As the herbivorous fecal matter is good source of anaerobic microbes, we have isolated Clostridium beijerinckiiYVU1(Acc.No.KP334151) and Clostridium saccharoperbutylacetonicumYVU2 (Acc. No. KP334152) strains from wild rabbit and optimized the fermentation factors that required for high yielding of butanol. Mango processing waste (mango peel and kernel flour) and sugarcane field residue (SFR) were selected as economically cheap substrates for the production ABE by the newly isolated solventogenic bacteria. The enzyme saccharification of mango peel resulted in 32% of reducing sugars release and 11.58 g/L butanol was produced by C.beijerinckiiYVU1, 10.5 g/L by the test culture, C. acetobutylicum2878 and 9.47 g/L by C. saccharoperbutylacetonicum YVU2. The production of butanol was 8.91g/L and 8.72g/L with respect to C.beijerinckiiYVU1 and C. saccharoperbutylacetonicumYVU2 from mango kernel flour. Novel sequential acid and alkali pretreatment effectively solubilized both the hemicelluloses and lignin and improved the glucose release (68%) from the cellulose in SFR.