A catabolite derepressed Bacillus subtilis strain KCC103 was used to produce α-amylase in medium containing sugarcane bagasse hydrolysate (SBH). Addition of SBH (1% reducing sugar (w/v)) to the nutrient medium supported maximum α-amylase production of 67.4 U ml-1. HPLC analysis of SBH showed the presence of glucose, xylose and arabinose in the ratio of 0.9:1.0:0.16 (w/w/w). In SBH-medium glucose and xylose were consumed completely while arabinose remained unutilized. Uptake rate of glucose was 2-folds higher than xylose but rate of α-amylase production with xylose was 1.5-folds higher than glucose. Arabinose had no effect on growth and α-amylase synthesis. Further, α-amylase production in SBH-medium was enhanced to 144.5 U ml-1 (2.2-fold) by response surface methodology where the levels of SBH, and other media components were varied. The modified medium consisted of (in g l-1) SBH: 24; peptone: 17.43; yeast extract: 1.32 and beef extract: 1.82. High level of SBH showed no significant inhibition of α-amylase synthesis. The derepressed strain KCC103 is useful to produce α-amylase economically in short time (30-36 h). © 2007 Elsevier Ltd. All rights reserved.