Purification of Natural Pigments Violacein and Deoxyviolacein Produced by Fermentation Using <i>Yarrowia lipolytica</i>
Georgio Nemer,
Nicolas Louka,
Paul Rabiller Blandin,
Richard G. Maroun,
Eugène Vorobiev,
Tristan Rossignol,
Jean-Marc Nicaud,
Erwann Guénin,
Mohamed Koubaa
Affiliations
Georgio Nemer
Université de Technologie de Compiègne, ESCOM, TIMR (Integrated Transformations of Renewable Matter), Centre de Recherche Royallieu—CS 60319, 60203 Compiègne CEDEX, France
Nicolas Louka
Laboratoire CTA, UR TVA, Centre d’Analyses et de Recherche, Faculté des Sciences, Université Saint-Joseph, Beyrouth 1104 2020, Lebanon
Paul Rabiller Blandin
Université de Technologie de Compiègne, ESCOM, TIMR (Integrated Transformations of Renewable Matter), Centre de Recherche Royallieu—CS 60319, 60203 Compiègne CEDEX, France
Richard G. Maroun
Laboratoire CTA, UR TVA, Centre d’Analyses et de Recherche, Faculté des Sciences, Université Saint-Joseph, Beyrouth 1104 2020, Lebanon
Eugène Vorobiev
Université de Technologie de Compiègne, ESCOM, TIMR (Integrated Transformations of Renewable Matter), Centre de Recherche Royallieu—CS 60319, 60203 Compiègne CEDEX, France
Tristan Rossignol
Université Paris-Saclay, INRAE, AgroParisTech, Micalis Institute, 78350 Jouy-en-Josas, France
Jean-Marc Nicaud
Université Paris-Saclay, INRAE, AgroParisTech, Micalis Institute, 78350 Jouy-en-Josas, France
Erwann Guénin
Université de Technologie de Compiègne, ESCOM, TIMR (Integrated Transformations of Renewable Matter), Centre de Recherche Royallieu—CS 60319, 60203 Compiègne CEDEX, France
Mohamed Koubaa
Université de Technologie de Compiègne, ESCOM, TIMR (Integrated Transformations of Renewable Matter), Centre de Recherche Royallieu—CS 60319, 60203 Compiègne CEDEX, France
Violacein and deoxyviolacein are bis-indole pigments synthesized by a number of microorganisms. The present study describes the biosynthesis of a mixture of violacein and deoxyviolacein using a genetically modified Y. lipolytica strain as a production chassis, the subsequent extraction of the intracellular pigments, and ultimately their purification using column chromatography. The results show that the optimal separation between the pigments occurs using an ethyl acetate/cyclohexane mixture with different ratios, first 65:35 until both pigments were clearly visible and distinguishable, then 40:60 to create a noticeable separation between them and recover the deoxyviolacein, and finally 80:20, which allows the recovery of the violacein. The purified pigments were then analyzed by thin-layer chromatography and nuclear magnetic resonance.