Altering the fatty acid profile of Yarrowia lipolytica to mimic cocoa butter by genetic engineering of desaturases

Abstract

Background Demand for Cocoa butter is steadily increasing, but the supply of cocoa beans is naturally limited and under threat from global warming. One route to meeting the future demand for cocoa butter equivalent (CBE) could be to utilize microbial cell factories such as the oleaginous yeast Yarrowia lipolytica. Results The main goal was to achieve triacyl-glycerol (TAG) storage lipids in Y. lipolytica mimicking cocoa butter. This was accomplished by replacing the native Δ9 fatty acid desaturase (Ole1p) with homologs from other species and changing the expression of both Ole1p and the Δ12 fatty acid desaturase (Fad2p). We thereby abolished the palmitoleic acid and reduced the linoleic acid content in TAG, while the oleic acid content was reduced to approximately 40 percent of the total fatty acids. The proportion of fatty acids in TAG changed dramatically over time during growth, and the fatty acid composition of TAG, free fatty acids and phospholipids was found to be very different. Conclusions We show that the fatty acid profile in the TAG of Y. lipolytica can be altered to mimic cocoa butter. We also demonstrate that a wide range of fatty acid profiles can be achieved while maintaining good growth and high lipid accumulation, which, together with the ability of Y. lipolytica to utilize a wide variety of carbon sources, opens up the path toward sustainable production of CBE and other food oils.

Urea is a drop-in nitrogen source alternative to ammonium sulphate in Yarrowia lipolytica

“…of the desaturases can massively alter the fatty acid profile. 26 , 32 , 33 Our results show that urea can be used as a drop-in nitrogen source instead of ammonium sulfate without massively altering the storage lipid profile.…”

“…food oils. 26 In these cases, nitrogen starvation is used to trigger lipid production to achieve high lipid titers. Under nitrogen-limiting conditions, the obese strain (OKYL049) showed significant changes in C16:0 and C18:2 between the two nitrogen sources.…”

Engineering Yarrowia lipolytica to produce nutritional fatty acids: Current status and future perspectives

“…Although there are still many barriers to settle for further improving the accumulation of nutritional fatty acids in Y. lipolytica , significant advances have been made. For example, many natural food substitutes, such as human milk fat substitutes and cocoa butter equivalents, have been produced by Y. lipolytica [ 96 , 97 ]. Nowadays, many emerging synthetic biology tools and gene editing technologies are on their way to become standard laboratory techniques for metabolic engineering of Y. lipolytica .…”

Tailoring and optimizing fatty acid production by oleaginous yeasts through the systematic exploration of their physiological fitness

“…A part of the previous studies on Y. lipolytica enabled switching the fatty acid composition by metabolic engineering strategies. These approaches were mainly focused on the manipulation of the elongation and desaturation pathway [ 30 , 31 ]. Additionally, Ochsenreither et al [ 19 ] reported that temperature and the composition of the medium lead to variation in fatty acid composition.…”