title: Conversion of cellulosic materials into glycolipid biosurfactants, mannosylerythritol lipids, by Pseudozyma spp. under SHF and SSF processes.
abstract: BACKGROUND
  Mannosylerythritol 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.
  
  
  RESULTS
  MEL 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 5048(T) or Pseudozyma aphidis PYCC 5535(T). 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.
  
  
  CONCLUSIONS
  This 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.
authors.affiliation: Department of Bioengineering and IBB - Institute for Bioengineering and Bioscience, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001, Lisboa, Portugal. nunosek@gmail.com.|MIT-Portugal Program, 77 Massachusetts Avenue, E40-221, Cambridge, MA, 02139, USA. nunosek@gmail.com.|Laboratório Nacional de Energia e Geologia, I.P, Unidade de Bioenergia, Estrada do Paço do Lumiar 22, 1649-038, Lisboa, Portugal. nunosek@gmail.com.
authors.forename: Nuno Torres
authors.initials: NT
authors.lastname: Faria
authors.affiliation: Department of Bioengineering and IBB - Institute for Bioengineering and Bioscience, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001, Lisboa, Portugal. marisaavsantos@tecnico.ulisboa.pt.|MIT-Portugal Program, 77 Massachusetts Avenue, E40-221, Cambridge, MA, 02139, USA. marisaavsantos@tecnico.ulisboa.pt.
authors.forename: Marisa
authors.initials: M
authors.lastname: Santos
authors.affiliation: Laboratório Nacional de Energia e Geologia, I.P, Unidade de Bioenergia, Estrada do Paço do Lumiar 22, 1649-038, Lisboa, Portugal. carlasofiarf@gmail.com.
authors.forename: Carla
authors.initials: C
authors.lastname: Ferreira
authors.affiliation: Laboratório Nacional de Energia e Geologia, I.P, Unidade de Bioenergia, Estrada do Paço do Lumiar 22, 1649-038, Lisboa, Portugal. susana.marques@lneg.pt.
authors.forename: Susana
authors.initials: S
authors.lastname: Marques
authors.affiliation: Department of Bioengineering and IBB - Institute for Bioengineering and Bioscience, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001, Lisboa, Portugal. frederico.ferreira@tecnico.ulisboa.pt.
authors.forename: Frederico Castelo
authors.initials: FC
authors.lastname: Ferreira
authors.affiliation: Laboratório Nacional de Energia e Geologia, I.P, Unidade de Bioenergia, Estrada do Paço do Lumiar 22, 1649-038, Lisboa, Portugal. cesar.fonseca@lneg.pt.
authors.forename: César
authors.initials: C
authors.lastname: Fonseca
mesh_terms: D002482:Cellulose / Q000378:metabolism*; D006017:Glycolipids / Q000096:biosynthesis*; D013501:Surface-Active Agents / Q000378:metabolism*; D014587:Ustilaginales / Q000254:growth & development*
chemical_list: D006017:Glycolipids; D013501:Surface-Active Agents; C439671:mannosylerythritol lipid; D002482:Cellulose