Identification of bifunctional Δ12/ω3 fatty acid desaturases for improving the ratio of ω3 to ω6 fatty acids in microbes and plants

We report the identification of bifunctional Δ12/ω3 desaturases fromFusarium moniliforme ,Fusarium graminearum , andMagnaporthe grisea . The bifunctional activity of these desaturases distinguishes them from all known Δ12 or ω3 fatty acid desaturases. The ω3 desaturase activity of these enzymes also shows a broad ω6 fatty acid substrate specificity by their ability to convert linoleic acid (LA), γ-linolenic acid, di-homo-γ-linolenic acid, and arachidonic acid to the ω3 fatty acids, α-linolenic acid (ALA), stearidonic acid, eicosatetraenoic acid, and eicosapentaenoic acid (EPA), respectively. Phylogenetic analysis suggests that ω3 desaturases arose by independent gene duplication events from a Δ12 desaturase ancestor. Expression ofF. moniliforme Δ12/ω3 desaturase resulted in high ALA content in bothYarrowia lipolytica , an oleaginous yeast naturally deficient in ω3 desaturation, and soybean. In soybean, seed-specific expression resulted in 70.9 weight percent of total fatty acid (%TFA) ALA in a transformed seed compared with 10.9%TFA in a null segregant seed and 53.2%TFA in the current best source of ALA, linseed oil. The ALA/LA ratio in transformed seed was 22.3, a 110- and 7-fold improvement over the null segregant seed and linseed oil, respectively. Thus, these desaturases have potential for producing nutritionally desirable ω3 long-chain polyunsaturated fatty acids, such as EPA, with a significantly improved ratio of ω3/ω6 long-chain polyunsaturated fatty acids in both oilseeds and oleaginous microbes.