Journal
MOLECULES
Volume 28, Issue 11, Pages -Publisher
MDPI
DOI: 10.3390/molecules28114292
Keywords
Yarrowia lipolytica; pigment; violacein; deoxyviolacein; fermentation; purification; column chromatography
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This study focuses on the biosynthesis of a mixture of violacein and deoxyviolacein using a genetically modified Y. lipolytica strain as a production chassis. The intracellular pigments were extracted and purified using column chromatography. The optimal separation between the pigments was achieved using an ethyl acetate/cyclohexane mixture with different ratios, followed by analysis using thin-layer chromatography and nuclear magnetic resonance.
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.
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