Production of carotenoids in oleaginous yeast and fungi
View Patent ↗The present invention provides systems for producing engineered oleaginous yeast or fungi that express carotenoids.
1. A recombinant oleaginous fungus selected from Yarrowia or Saccharomyces characterized by:
i. lipid accumulation to at least 20% of its dry weight; and
ii. as a result of a recombinant process, production of at least one carotenoid and accumulation of the at least one carotenoid to at least 1% of its dry cell weight, wherein the at least one carotenoid is selected from the group consisting of beta-carotene and astaxanthin.
2. The recombinant fungus of claim 1 , wherein the lipid accumulation to at least 20% of its dry cell weight occurs as a result of genetic engineering.
3. The recombinant fungus of claim 2 , wherein the fungus is selected from a species group consisting of Yarrowia lipolytica and Saccharomyces cerevisiae.
4. The recombinant fungus of claim 1 , wherein the at least one carotenoid is accumulated in cytoplasmic oil bodies.
5. The recombinant fungus of claim 1 , wherein the recombinant process alters the expression or activity of a carotenogenic polypeptide selected from the group consisting of isoprenoid biosynthesis polypeptides, carotenoid biosynthesis polypeptides, isoprenoid biosynthesis competitor polypeptides, and combinations thereof.
6. The recombinant fungus of claim 5 , wherein the carotenoid biosynthesis polypeptide is selected from the group consisting of: phytoene synthase polypeptide, phytoene dehydrogenase polypeptide, lycopene cyclase polypeptide, carotenoid ketolase polypeptide, carotenoid hydroxylase polypeptide, astaxanthin synthase polypeptide, carotenoid epsilon hydroxylase polypeptide, carotenoid glucosyltransferase polypeptide, lycopene cyclase (beta and epsilon subunits) polypeptides, and acyl CoA:diacyglycerol acyltransferase polypeptide.
7. The genetically recombinant fungus of claim 5 , wherein the carotenogenic polypeptide confers to the yeast the ability to produce the at least one carotenoid to a level at least 1% of its dry cell weight or confers to the yeast the ability to produce at least one carotenoid which the yeast does not naturally produce.
8. The recombinant fungus of claim 5 , wherein the fungus comprises at least one modification selected from the group consisting of:
i. expression of truncated HMG-CoA reductase;
ii. heterologous expression of carotene hydroxylase and carotene ketolase;
iii. complete or partial inactivation of a gene encoding squalene synthase; and
iv. combinations thereof.
9. The recombinant fungus of claim 1 , further characterized by an oleaginic modification.
10. The recombinant fungus of claim 9 , wherein the oleaginic modification confers upon the fungus an ability to accumulate lipid to at least 20% of its dry cell weight.
11. The recombinant fungus of claim 9 , wherein the oleaginic modification alters expression or activity of at least one oleaginic polypeptide.
12. The recombinant fungus of claim 11 , wherein the at least one oleaginic polypeptide is selected from the group consisting of acetyl-CoA carboxylase polypeptide, pyruvate decarboxylase polypeptide, isocitrate dehydrogenase polypeptide, ATP citrate lyase polypeptide, malic enzyme polypeptide, AMP deaminase polypeptide, and combinations thereof.