High arachidonic acid producing strains of yarrowia lipolytica

Abstract

Engineered strains of the oleaginous yeast Yarrowia lipolytica capable of producing greater than 10% arachidonic acid (ARA, an ω-6 polyunsaturated fatty acid) in the total oil fraction are described. These strains comprise various chimeric genes expressing heterologous desaturases, elongases and acyltransferases, and optionally comprise various native desaturase and acyltransferase knockouts to enable synthesis and high accumulation of ARA. Production host cells are claimed, as are methods for producing ARA within said host cells.

1 - 14 . (canceled) 
     
     
         15 . A recombinant oleaginous  Yarrowia  yeast cell for the production of arachidonic acid comprising a background  Yarrowia  sp. comprising:
 a) at least one gene encoding Δ9 elongase; and   b) at least one gene encoding Δ8 desaturase;   wherein the oleaginous  Yarrowia  yeast cell accumulates in excess of about 25% of its dry cell weight as oil and the oleaginous  Yarrowia  yeast cell produces at least about 14% arachidonic acid of total lipids produced.   
     
     
         16 - 19 . (canceled) 
     
     
         20 . A microbial oil produced by a method comprising:
 a) culturing the recombinant oleaginous  Yarrowia  yeast cell of  claim 15  wherein a microbial oil comprising arachidonic acid is produced; and   b) optionally, recovering the microbial oil of step (a).   
     
     
         21 . The microbial oil of  claim 20  wherein the oil contains at least about 5% arachidonic acid. 
     
     
         22 . The microbial oil of  claim 20  blended with an oil comprising a fatty acid selected from the group consisting of linoleic acid, γ-linolenic acid, eicosadienoic acid, dihomo-γ-linolenic acid, arachidonic acid, α-linolenic acid, stearidonic acid, eicosatrienoic acid, eicosatetraenoic acid, eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid. 
     
     
         23 . A food product comprising an effective amount of the microbial oil of  claim 20 . 
     
     
         24 . A food product according to  claim 23  selected from the group consisting of a food analogue, a meat product, a cereal product, a baked food, a snack food and a dairy product. 
     
     
         25 . A product selected from the group consisting of a medical food, a dietary supplement, an infant formula and a pharmaceutical comprising an effective amount of the microbial oil of  claim 20 . 
     
     
         26 . An animal feed comprising an effective amount of the microbial oil of  claim 20 . 
     
     
         27 . An animal feed according to  claim 26  wherein the animal feed is selected from the group consisting of a pet feed, a ruminant feed, a poultry feed and an aquacultural feed. 
     
     
         28 . An animal feed comprising an effective amount of microbial oil and optionally comprising a yeast biomass comprising the recombinant oleaginous  Yarrowia  yeast cell of  claim 15 . 
     
     
         29 . An animal feed according to  claim 28  wherein the yeast biomass comprises feed nutrients selected from the group consisting of proteins, lipids, carbohydrates, vitamins, minerals and nucleic acids. 
     
     
         30 - 36 . (canceled)