Highly efficient fed-batch modes for enzymatic hydrolysis and microbial lipogenesis from alkaline organosolv pretreated corn stover for biodiesel production

High-density culture for microbial lipid preparation from low-cost lignocellulosic feedstocks is crucial for commercial-scale biodiesel production. Herein, fed-batch saccharification of alkaline organosolv pretreatment (AOP) of corn stover at an extremely high solids content of 47% (w/v) released 299.5 g/L of lignocellulosic sugars including 18.3% of soluble oligosaccharides. Three types of liquid hydrolysates for seed culture, fermentation, and feeding during fed-batch culture were obtained from the hydrolysate slurry using a two-step washing strategy with 99.3% of sugars recovery. Cutaneotrichosporon oleaginosum showed excellent capacity for assimilating both monosaccharides and oligosaccharides for lipid production using the fed-batch culture mode. Lipid concentration, content, and yield gained 42.3 g/L, 64.6%, and 20.4 g/100 g, respectively. Turbid hydrolysate collected with high recovery of high-concentration sugars and simplified process could be directly served as feeding medium. In general, the overall hydrolysis yield and lipid yield using fed-batch mode accounted for 93.2% and 97.6% of those using batch mode, respectively, resulting in a lipid output of 102.8 g/kg raw corn stover. The fatty acid composition and the prediction of biodiesel properties of lipid samples indicated the suitability for high-quality fuel production. This study provided valuable information for designing highly efficient lignocelluloses-to-biodiesel routes.