| Title: | Biosynthesis of insect sex pheromone precursors via engineered beta-oxidation in yeast |
| Author(s): | Petkevicius K; Wenning L; Kildegaard KR; Sinkwitz C; Smedegaard R; Holkenbrink C; Borodina I; |
| Address: | "The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet 220, 2800 Kgs. Lyngby, Denmark. BioPhero ApS, Lerso Parkalle 42-44, 4th, 2100 Copenhagen O, Denmark" |
| ISSN/ISBN: | 1567-1364 (Electronic) 1567-1356 (Print) 1567-1356 (Linking) |
| 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 beta-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 11 peroxisomal oxidases of different origins. Yeast cultivations were performed with supplementation of methyl myristate (14:Me). The oxidase Lbo_31670 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 beta-oxidation in Y. lipolytica allowed expanding the portfolio of microbially produced insect sex pheromones" |
| Keywords: | Amino Acid Sequence Animals Coenzyme A/metabolism Drosophila melanogaster/genetics Fatty Acid Desaturases/genetics/metabolism Insecta *Moths Myristates/metabolism Oxidoreductases/genetics/metabolism *Sex Attractants/genetics/metabolism Yeasts/genetics Yar; |
| Notes: | "MedlinePetkevicius, Karolis Wenning, Leonie Kildegaard, Kanchana R Sinkwitz, Christina Smedegaard, Rune Holkenbrink, Carina Borodina, Irina eng Research Support, Non-U.S. Gov't England 2022/08/11 FEMS Yeast Res. 2022 Sep 1; 22(1):foac041. doi: 10.1093/femsyr/foac041" |
