This study investigated the co-fermentation of molasses and sweetpotato vine hydrolysate (SVH) by Trichosporon fermentans. T. fermentans showed low lipid accumulation on pure molasses; however, its lipid content increased by 35% when 10% SVH was added. The strong influence of SVH on lipid production was further demonstrated by the result of sensitivity analysis on effects of factors based on an artificial neural network model because the relative importance value of SVH dosage for lipid production was only lower than that of fermentation time. Scanning electron microscope observation and flow cytometry of yeast cells grown in culture with and without SVH showed that less deformation cells were involved in the culture with SVH. The activity of malic enzyme, which plays a key role in fatty acid synthesis, increased from 2.4 U/mg to 3.7 U/mg after SVH added. All results indicated SVH is a good supplement for lipid fermentation on molasses. (C) 2014 Elsevier Ltd. All rights reserved.