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At 10μgml-1 of 2,4-D, the highest starch concentration of 989μgmg-1was achieved in C. humicola. Finally, starch in C. humicola were hydrolysed and ABE fermentation was performed using C. acetobutylicum under anaerobic condition in a 5 L automated fermenter. After 72 h of fermentation, the fermented broth is analyed in Gas Chromatography showing the fermented product containing Acetone Butanol Ethanol. The present study is the first report on the production of biobutanol from C. humicola isolated from Temple pond. This study emphasizes