Enzymatic hydrolysate of sake lees containing glucose. succinic acid, formic acid and ethanol was used as a substrate for succinic acid production by Actinobacillus succinogenes 130Z. Results indicated that the available nutrients in enzymatic hydrolysate of sake lees could be able to overcome the inhibitory effect in succinic acid fermentation. By optimizing the culture medium for succinic acid fermentation with enzymatic hytholysate of sake lees, the nitrogen sources could be reduced from 10 g/L yeast extract and 8 g/L corn steep liquor to just 2.5 g/L yeast extract. When batch fermentation was carried out in a 3-L fermentor using enzymatic hydrolysate of sake lees with 2.5 g/L yeast extract and the salts mixture, 48.0 g/L succinic acid was obtained with consumption of 63.6 g/L glucose in enzymatic hydrolysate of sake lees at 51 h. This study suggests that enzymatic hydrolysate of sake lees may be an alternative substrate for the efficient production of succinic acid by A. succinogenes 130Z. (c) 2010 Elsevier Inc. All rights reserved.