Production of Long Chain Fatty Alcohols Found in Bumblebee Pheromones by Yarrowia lipolytica

Abstract

Fatty alcohols (FA-OH) are aliphatic unbranched primary alcohols with a chain of four or more carbon atoms. Besides potential industrial applications, fatty alcohols have important biological functions as well. In nature, fatty alcohols are produced as a part of a mixture of pheromones in several insect species, such as moths, termites, bees, wasps, etc. In addition, FA-OHs have a potential for agricultural applications, for example, they may be used as a suitable substitute for commercial insecticides. The insecticides have several drawbacks associated with their preparation, and they exert a negative impact on the environment. Currently, pheromone components are prepared mainly through the catalytic hydrogenation of plant oils and petrochemicals, which is an unsustainable, ecologically unfriendly, and highly expensive process. The biotechnological production of the pheromone components using engineered microbial strains and through the expression of the enzymes participating in the biosynthesis of these components is a promising approach that ensures ecological sustenance as well. The present study was aimed at evaluating the production of FA-OHs in the oleaginous yeast, Yarrowia lipolytica, with different lengths of fatty-acyl chains by expressing the fatty acyl-CoA reductase (FAR) BlapFAR4 from B. lapidarius, producing C16:0-OH, C16:1Δ9-OH, and lower quantities of both C14:0-OH and C18:1Δ9-OH, and BlucFAR1 from B. lucorum, producing FA-OHs with a chain length of 18–26 carbon atoms, in this yeast. Among the different novel Y. lipolytica strains used in the present study, the best results were obtained with JMY7086, which carried several lipid metabolism modifications and expressed the BlucFAR1 gene under the control of a strong constitutive promoter 8UAS-pTEF. JMY7086 produced only saturated fatty alcohols with chain lengths from 18 to 24 carbon atoms. The highest titer and accumulation achieved were 166.6 mg/L and 15.6 mg/g DCW of fatty alcohols, respectively. Unlike JMY7086, the BlapFAR4-expressing strain JMY7090 produced only 16 carbon atom-long FA-OHs with a titer of 14.6 mg/L.

Enhancing very long chain fatty acids production in Yarrowia lipolytica

“…E. coli strain EC61 (JMP62 -8UAS-pTEF-TaFAE1-LEU2ex ) was constructed in this work. Plasmids (JMP62 -pTEF-TaFAE1-LEU2ex ) from the E. coli EC11 [ 14 ] and (JMP62 -URA3ex-8UASpTEF-FAR4 ) from E. coli JME4305 [ 41 ] were double-digested by ClaI and BamHI to replace pTEF promoter in (JMP62 -pTEF-TaFAE1-LEU2ex ) by 8UAS-pTEF from (JMP62 -URA3ex-8UAS-pTEF-FAR4 ). The resulting plasmid (JMP62- 8UAS-pTEF-TaFAE1-LEU2ex ) was stored in E. coli EC61.…”

Developments in Fatty Acid-Derived Insect Pheromone Production Using Engineered Yeasts

“…In another study, expression of the gene encoding FAR, BlapFAR 4 from Bombus lapidarius or BlucFAR1 from Bombus lucorum in Y. lipolytica , led to the production of bumblebee pheromones consisting of long-chain fatty alcohols. The titer of saturated fatty alcohols with C18-C24 was 166.6 mg/L, while the titer of C16 FA-OHs (C16:0-OH and C16:1-OH) was 14.6 mg/L ( Table 1 ; Hambalko et al, 2020 ). However, in S. cerevisiae , expression of BlucFAR1 produced only a small amount of FA-OHs (6.9 mg/L), while the expression of BlapFAR4 led to the production of 79 mg/L of C16-OHs ( Table 1 ; Tupec et al, 2019 ).…”

Improving methanol assimilation in Yarrowia lipolytica via systematic metabolic engineering combined with compartmentalization

“…13 In this regard, baker's yeast (Saccharomyces cerevisiae), and some nonconventional yeasts with specific features of interest, such as high secretion ability or lipophilic compound accumulation (Yarrowia lipolytica), are regarded as desirable chassis cells for the construction of synthetic methylotrophs. [16][17][18]39 S. cerevisiae had first been constructed as a methylotrophic yeast by equipping a methanol assimilation module, which led to 0.26 g L −1 pyruvate using methanol and the yeast extract as a co-substrate. 14,15 More recently, Y. lipolytica has been engineered for methanol assimilation by construction of a chimeric assimilation pathway, rewiring of the native precursor supply pathway and laboratory evolution.…”