DI-UMONS : Dépôt institutionnel de l’université de Mons

Recherche transversale
Rechercher
(titres de publication, de périodique et noms de colloque inclus)
2019-05-13 - Colloque/Présentation - communication orale - Anglais - page(s)

Delfau-Bonnet Guillaume , Clément Tiphaine, Allais Florent, Hantson Anne-Lise , "Lipids tracking by flow cytometer in four oleaginous yeasts" in Advanced World Research on Yeast, Tokyo, Japon, 2019

  • Codes CREF : Biotechnologie (DI3800), Fermentation biosynthèse (DI3810)
  • Unités de recherche UMONS : Génie des Procédés chimiques et biochimiques (F505)
  • Instituts UMONS : Institut de Recherche en Science et Ingénierie des Matériaux (Matériaux), Institut des Biosciences (Biosciences)

Abstract(s) :

(Anglais) Oleaginous yeasts are able to metabolize different sources of carbon (e.g., monosaccharides, disaccharides, glycerol) into various kinds of lipids (1). Oil from yeasts can be used for different applications such as food, pharmaceutical or third generation biofuels, but interest is growing for the production of long chain poly-unsaturated fatty acids as building blocks for the polymer. Yeasts lipids extraction and quantification is a fastidious and time-consuming process involving several steps and the use of non-environmentally friendly solvents such as hexane and methanolic chloride 3M. The classical method involves lyophilization of biomass, trans-methylation in methanolic chloride and extraction in heptane. An alternative method to assess yeasts lipids production during fermentation consists in dyeing intracellular lipids with a fluorochrome. The most popular lipid dye is Nile red, which binds non- specifically neutral lipids. The BODIPY 493/503 dye is more specific to neutral lipid than Nile red (3). In this study, four oleaginous yeast strains (two strains of Yarrowia lipolytica clib 89 and clib 718, one Lipomyces starkeyi mucl 27779 and one Meyerozyma guillermondii clib 222) were compared for their ability to accumulate lipids during growth. Intracellular lipids accumulation was monitored during yeasts growth, both by Bligh and Dyer protocol and flow cytometry after cell dyeing with Nile red or BODIPY 493/503. GC analysis show that Yarrowia lipolytica clib 89 and clib 718 accumulated more lipids in total but Lipomyces starkeyi presented the highest production of α-linolenic acid and linoleic acid, a long chain polyunsatured acid of interest for the chemical industry. Flow cytometry analysis with BODIPY 493/503 marking (dyeing?) showed a good correlation of fluorescence intensity with both total lipid accumulation and polyunsaturated fatty acid concentration. Yeast marking during fermentation with BODIPY 493/503 associated with flow cytometry is thus a very promising technique to measure and compare the ability of yeasts to synthesize and accumulate lipids, in particular those containing long chain polyunsaturated fatty acids.