a b s t r a c tThe pentose-assimilating capacity of yeasts from the genus Pseudozyma, P. antarctica PYCC 5048 T , P. aphidis PYCC 5535 T and P. rugulosa PYCC 5537 T , was exploited towards the production of mannosylerythritol lipids (MEL), a glycolipid with biosurfactant properties. The three strains tested were able to grow on d-xylose and l-arabinose with similar maximum specific growth rates to those estimated on d-glucose (around 0.2 h −1 ). The highest MEL titres (4.8-5.4 g/l) and yields (0.11-0.14 g/g) from d-xylose were found in P. antarctica PYCC 5048 T , which presented similar values to those estimated on d-glucose and on d-xylose/d-glucose mixtures. P. rugulosa PYCC 5537T showed a pattern of sugar conversion into MEL similar to P. antarctica, but at 40% lower titres. P. aphidis PYCC 5535 T presented lower MEL titres from d-xylose (1.2 g/l) than from d-glucose (3.4 g/l). Nitrate supply increased sugar consumption rate and, when accompanied by d-glucose or d-xylose feeding, additional biomass production. In this case, sugar was completely consumed before sugar feeding at day 7, but not when feeding is performed at day 4. Higher MEL titres where obtained for the later condition reaching 7.3 g/l and 5.8 g/l, in fed-batch cultures with glucose and xylose, respectively.

BackgroundMannosylerythritol lipids (MEL) are glycolipids with unique biosurfactant properties and are produced by Pseudozyma spp. from different substrates, preferably vegetable oils, but also sugars, glycerol or hydrocarbons. However, solvent intensive downstream processing and the relatively high prices of raw materials currently used for MEL production are drawbacks in its sustainable commercial deployment. The present work aims to demonstrate MEL production from cellulosic materials and investigate the requirements and consequences of combining commercial cellulolytic enzymes and Pseudozyma spp. under separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) processes.ResultsMEL was produced from cellulosic substrates, Avicel® as reference (>99% cellulose) and hydrothermally pretreated wheat straw, using commercial cellulolytic enzymes (Celluclast 1.5 L® and Novozyme 188®) and Pseudozyma antarctica PYCC 5048T or Pseudozyma aphidis PYCC 5535T. The strategies included SHF, SSF and fed-batch SSF with pre-hydrolysis. While SSF was isothermal at 28°C, in SHF and fed-batch SSF, yeast fermentation was preceded by an enzymatic (pre-)hydrolysis step at 50°C for 48 h. Pseudozyma antarctica showed the highest MEL yields from both cellulosic substrates, reaching titres of 4.0 and 1.4 g/l by SHF of Avicel® and wheat straw (40 g/l glucan), respectively, using enzymes at low dosage (3.6 and 8.5 FPU/gglucan at 28°C and 50°C, respectively) with prior dialysis. Higher MEL titres were obtained by fed-batch SSF with pre-hydrolysis, reaching 4.5 and 2.5 g/l from Avicel® and wheat straw (80 g/l glucan), respectively.ConclusionsThis work reports for the first time MEL production from cellulosic materials. The process was successfully performed through SHF, SSF or Fed-batch SSF, requiring, for maximal performance, dialysed commercial cellulolytic enzymes. The use of inexpensive lignocellulosic substrates associated to straightforward downstream processing from sugary broths is expected to have a great impact in the economy of MEL production for the biosurfactant market, inasmuch as low enzyme dosage is sufficient for good systems performance.