In this work, hydrolysis of cellulose and hemicellulose content of palm kernel cake (PKC) by different types of hydrolytic enzymes was studied to evaluate monomeric sugars released for production of biobutanol by Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564) in acetone-butanol-ethanol (ABE) fermentation. Experimental results revealed that when PKC was hydrolyzed by mixed beta-glucosidase, cellulase and mannanase, a total simple sugars of 87.81 +/- 4.78 g/L were produced, which resulted in 3.75 +/- 0.18 g/L butanol and 6.44 +/- 0.43 g/L ABE at 168 h fermentation. In order to increase saccharolytic efficiency of enzymatic treatment, PKC was pretreated by liquid hot water before performing enzymatic hydrolysis. Test results showed that total reducing sugars were enhanced to 97.81 +/- 1.29 g/L with elevated production of butanol and ABE up to 4.15 +/- 1.18 and 7.12 +/- 2.06 g/L, respectively which represented an A: B: E ratio of 7: 11: 1. (C) 2015 Elsevier Ltd. All rights reserved.