Production of sterols in oleaginous yeast and fungi

A recombinant fungus characterized by:

• a. the recombinant fungus is oleaginous in that it can accumulate lipid to at least about 20% of its dry cell weight;
• and
• b. the recombinant fungus produces at least one sterol compound, and can accumulate the produced sterol compound to at least about 1% of its dry cell weight;
• wherein the recombinant fungus is a member of a genus selected from the group consisting of:
• Aspergillus, Blakeslea, Botrytis, Candida, Cercospora, Cryptococcus, Cunninghamella, Fusarium (Gibberella), Kluyveromyces, Lipomyces, Mortierella, Mucor, Neurospora, Penicillium, Phycomyces, Pichia (Hansenula), Puccinia, Pythium, Rhodosporidium, Rhodotorula, Saccharomyces, Sclerotium, Trichoderma, Trichosporon, Xanthophyllomyces (Phaffia), and Yarrowia;
• or is a species selected from the group consisting of:
• Aspergillus terreus, Aspergillus nidulans, Aspergillus niger, Blakeslea trispora, Botrytis cinerea, Candida japonica, Candida pulcherrima, Candida revkaufi, Candida tropicalis, Candida utilis, Cercospora nicotianae, Cryptococcus curvatus, Cunninghamella echinulata, Cunninghamella elegans, Fusarium fujikuroi (Gibberella zeae), Kluyveromyces lactis, Lipomyces starkeyi, Lipomyces lipoferus, Mortierella alpina, Mortierella ramanniana, Mortierella isabellina, Mortierella vinacea, Mucor circinelloides, Neurospora crassa, Phycomyces blakesleanus, Pichia pastoris, Puccinia distincta, Pythium irregulare, Rhodosporidium toruloides, Rhodotorula glutinis, Rhodotorula graminis, Rhodotorula mucilaginosa, Rhodotorula pinicola, Rhodotorula gracilis, Saccharomyces cerevisiae, Sclerotium rolfsii, Trichoderma reesei, Trichosporon cutaneum, Trichosporon pullans, Xanthophyllomyces dendrorhous (Phaffia rhodozyma), and Yarrowia lipolytica;
• wherein the recombinant fungus comprises at least one modification as compared with a parental fungus, the at least one modification being selected from the group consisting of sterologenic modifications, oleaginic modifications, and combinations thereof, and
• wherein the at least one modification alters oleaginicity of the recombinant fungus, confers to the recombinant fungus oleaginy, confers to the recombinant fungus the ability to produce the at least one sterol compound to a level at least about 1% of its dry cell weight, or confers to the recombinant fungus the ability to produce at least one sterol compound which the parental fungus does not naturally produce.

An engineered Y. lipolytica strain that produces at least one sterol compound selected from the group consisting of:

• squalene, lanosterol, zymosterol, ergosterol or 7-dehydrocholesterol (provitamin D3), the strain containing one or more sterologenic modifications selected from the group consisting of:
• a. decreased expression or activity of a Y. lipolytica GGPP synthase polypeptide;
• b. increased expression or activity of a truncated HMG CoA reductase polypeptide;
• c. increased expression or activity of an FPP synthase polypeptide;
• d. increased expression or activity of an IPP isomerase polypeptide;
• e. increased expression or activity of an HMG-CoA synthase polypeptide;
• f. increased expression or activity of a mevalonate kinase polypeptide;
• g. increased expression or activity of a phosphomevalonate kinase polypeptide;
• h. increased expression or activity of a mevalonate pyrophosphate decarboxylate polypeptide;
• i. increased expression or activity of a cytosolic malic enzyme polypeptide;
• j. increased expression or activity of a malate dehydrogenase polypeptide;
• k. increased expression or activity of an AMP deaminase polypeptide;
• l. increased expression or activity of a glucose 6 phosphate dehydrogenase polypeptide;
• m. increased expression or activity of a malate dehydrogenase homolog 2 polypeptide;
• n. increased expression or activity of a GND1-6-phosphogluconate dehydrogenase polypeptide;
• o. increased expression or activity of a isocitrate dehydrogenase polypeptide;
• p. increased expression or activity of a IDH2-isocitrate dehydrogenase polypeptide;
• q. increased expression or activity of a fructose 1,6 bisphosphatase polypeptide;
• r. increased expression or activity of a Erg10-acetoacetyl CoA thiolase polypeptide;
• s. increased expression or activity of a squalene synthase polypeptide;
• and combinations thereof.