Scale-up of two-step acid-catalysed glycerol pretreatment for production of oleaginous yeast biomass from sugarcane bagasse by Rhodosporidium toruloides.

Two-step dilute acid and acid-catalysed glycerol pretreatment was developed to maximise sugar yield from sugarcane bagasse. At the laboratory scale, dilute acid pretreatment at 130 °C followed by acid-catalysed glycerol pretreatment at 170 °C led to a total sugar (C5 + C6) yield of 82%, 31% higher than that from one-step acid-catalysed glycerol pretreatment. At the pilot scale, the two-step dilute acid and acid-catalysed glycerol pretreatment led to a maximum sugar yield of 74%, 13% higher than that from one-step pretreatment with 52% reduction in glycerol usage. The enzymatic hydrolysate containing glucose and residual glycerol were used to produce microbial oils by a Rhodosporidium toruloides strain. A fed-batch cultivation strategy led to the production of 44.8 g/L cell mass, including 26.6 g/L oil, 8.6 g/L protein and 12.7 mg/L carotenoid. The cell mass and oil yields were 19% higher than those from batch cultivation as feedstock inhibition and catabolite repression were alleviated.