Biosynthesis of insect sex pheromone precursors via engineered β-oxidation in yeast

Abstract

Mating disruption with insect sex pheromones is an attractive and environmentally friendly technique for pest management. Several Lepidoptera sex pheromones have been produced in yeast, where biosynthesis could be accomplished by the expression of fatty acyl-CoA desaturases and fatty acyl-CoA reductases. In this study, we aimed to develop yeast Yarrowia lipolytica cell factories for producing Lepidoptera pheromones which biosynthesis additionally requires β-oxidation, such as (Z)-7-dodecenol (Z7-12:OH), (Z)-9-dodecenol (Z9-12:OH) and (Z)-7-tetradecenol (Z7-14:OH). We expressed fatty acyl-CoA desaturases from Drosophila melanogaster (Dmd9) or Lobesia botrana (Lbo_PPTQ) and fatty acyl-CoA reductase from Helicoverpa armigera (HarFAR) in combinations with eleven peroxisomal oxidases of different origins. Yeast cultivations were performed with supplementation of methyl myristate (14:Me). The oxidase Lbo_31 670 from L. botrana provided the highest titers of (Z)-7-dodecenoate, (Z)-9-dodecenoate, and (Z)-7-tetradecenoate. However, no chain-shortened fatty alcohols were produced. The mutation of fatty acid synthase (Fas2pI1220F) to increase myristate production did not lead to targeted fatty alcohol production. The problem was solved by directing the reductase into peroxisomes, where the strain with Dmd9 produced 0.10 ± 0.02 mg/L of Z7-12:OH and 0.48 ± 0.03 mg/L of Z7-14:OH, while the strain with Lbo_PPTQ produced 0.21 ± 0.03 mg/L of Z9-12:OH and 0.40 ± 0.07 mg/L of Z7-14:OH. In summary, the engineering of β-oxidation in Y. lipolytica allowed expanding the portfolio of microbially produced insect sex pheromones.

Mid–Long Chain Dicarboxylic Acid Production via Systems Metabolic Engineering: Progress and Prospects

“…Notably, ACO is the rate-limiting enzyme in the first step of β-oxidation . A multifunctional enzyme (MFE2) catalyzes the second and third steps of β-oxidation, wherein trans-2-enoyl-CoA is oxidized to 3-ketoacyl-CoA . Analyses of gene regulatory mechanisms have revealed the presence of a glucose action element and an oleic acid action element in the regions 393–526 bp and 341–393 bp upstream of the ACO-coding region.…”

“…54 A multifunctional enzyme (MFE2) catalyzes the second and third steps of β-oxidation, wherein trans-2-enoyl-CoA is oxidized to 3-ketoacyl-CoA. 55 Analyses of gene regulatory mechanisms have revealed the presence of a glucose action element and an oleic acid action element in the regions 393− 526 bp and 341−393 bp upstream of the ACO-coding region. Hence, the expression level of the ACO gene can be modulated under different carbon source induction conditions.…”

Identification and characterisation ofPlanococcus citri cis- andtrans-isoprenyl diphosphate synthase genes, supported by short- and long-read transcriptome data

“…Alternative management strategies, such as mating disruption, demand larger quantities and, therefore, require low-cost, scalable production methods (Zou and Millar, 2015). Biotechnological production of sex pheromones has been demonstrated recently for lepidopteran pheromones (Mateos-Fernández et al, 2021; Petkevicius et al, 2022) and biotechnological synthesis has been proposed as a sustainable route for large-scale, low-cost pheromone production. However, these methods require knowledge of the biosynthetic pathways.…”

Independent evolution of the iridoid biosynthetic pathway in plants and insects

“…For the management of pests, away from the use of broad-spectrum eradicant pesticides and their rapid development of resistance, pheromones are now being realized as more sustainable alternatives, e.g., the leading pesticide manufacturer FMC’s takeover for $200 million of BioPhero ( 10 ). FMC is thereby developing the production of nonisoprenoidal insect pheromones in commercial competition with conventional pesticides by expressing genes for insect sex pheromone biosynthesis in engineered yeasts for fermenter production ( 11 ). However, the next wider goal will be to do similarly for isoprenoidal pheromones and other signal chemicals (semiochemicals) which can again benefit from the work here ( 5 ) with the choice of iridoid synthesis genes across kingdoms and other taxonomic divisions ( Fig.…”