This work describes an innovative process for the extraction of lycopene from mated cultures of Blakeslea trispora using supercritical carbon dioxide extraction (SCE) containing acetone as an entrainer. The results obtained are compared to conventional solvent extraction. Incorporation of entrainer improved the recovery of lycopene from 73% to 85% by SCE. Box-Behnken design was used to optimize the operating conditions for maximum extraction of lycopene from the biomass. The parameters optimized were temperature (45-65 degrees C), pressure (300-400 bar), and extraction time (1.0-2.0 h) at a fixed flow rate of 1 ml/ min of carbon dioxide and entrainer at 1 ml/g biomass. The optimum conditions were found to be 1.1 h extraction at 52 degrees C and 349 bar, resulting in a 92% recovery of lycopene. The use of supercritical fluid extraction along with an entrainer is therefore recommended as a better option for enhancing the recovery of lycopene from B. trispora. (C) 2008 Elsevier Ltd. All rights reserved.