Microalga Chlorella protothecoides can grow heterotrophically with glucose as the carbon source and accumulate high proportion of lipids. The microalgal lipids are suitable for biodiesel production. To further increase lipid yield and reduce biodiesel cost, sweet sorghum juice was investigated as an alternative carbon source to glucose in the present study. When the initial reducing sugar concentration was 10 g L(-1) in the culture medium, the dry cell yield and lipid content were 5.1 g L(-1) and 52.5% using enzymatic hydrolyzates of sweet sorghum juice as the carbon source after 120 h-culture in flasks. The lipid yield was 35.7% higher than that using glucose. When 3.0 g L(-1) yeast extract was added to the medium, the dry cell yield and lipid productivity was increased to 1.2 g L(-1) day(-1) and 586.8 mg L(-1) day(-1). Biodiesel produced from the lipid of C. protothecoides through acid catalyzed transesterification was analyzed by GC-MS, and the three most abundant components were oleic acid methyl ester, cetane acid methyl ester and linoleic acid methyl ester. The results indicate that sweet sorghum juice could effectively enhance algal lipid production, and its application may reduce the cost of algae-based biodiesel. (C) 2009 Elsevier Ltd. All rights reserved.