Compositions and methods for type iii polyketide production in oleaginous yeast species

Abstract

This invention relates to compositions and methods for the production of Type III polyketides using genetically modified oleaginous yeast strains, for example, Yarrowia lipolytica.

1 .- 72 . (canceled) 
     
     
         73 . A method for production of a Type III polyketide comprising:
 a. culturing a genetically modified oleaginous yeast cell comprising a gene encoding Acetyl-CoA Carboxylase (AceCarb (ACC1); YALI0C11407g), wherein the oleaginous yeast cell is  Yarrowia lipolytica , and wherein the oleaginous yeast cell further comprises the following modification: an overexpression of peroxin 10 (PEX10; YALI0C01023) in a growth medium; and   b. isolating the Type III polyketide.   
     
     
         74 . The method of  claim 73 , wherein the oleaginous yeast cell further comprises a heterologous gene encoding a heterologous Type III polyketide synthase, further wherein the Type III polyketide synthase is selected from the group consisting of a CAA86219.2 gene from  Gerbera hybrida , AY517486 gene from  Rheum palmatum , JX840717-JX840724 gene from  Pseudomonas fluorescens , B0LDU5 gene from  Rubus idaeus , B1VLQ8 gene from  Streptomyces griseus , SCO1206 from  Streptomyces coelicolor , BAB12102.2 gene from  Humulus lupulus , or a combination thereof. 
     
     
         75 . The method of  claim 73 , wherein the gene is incorporated into a gene expression cassette comprising a promoter and the gene, wherein the gene expression cassette is integrated into a genome of the yeast cell. 
     
     
         76 . The method of  claim 73 , wherein at least two copies of the gene are present in a genome of the yeast cell. 
     
     
         77 . The method of  claim 73 , wherein the gene is episomally expressed from a plasmid. 
     
     
         78 . The method of  claim 73 , wherein the yeast cell comprises an additional genetic modification. 
     
     
         79 . The method of  claim 78 , wherein the additional genetic modification increases the acetyl-CoA or malonyl-CoA levels or fluxes. 
     
     
         80 . The method of  claim 78 , wherein the additional genetic modification increases the rate of beta-oxidation to increase the acetyl-CoA or malonyl-CoA levels or fluxes. 
     
     
         81 . The method of  claim 73 , wherein the yeast cell is a PO1f strain. 
     
     
         82 . The method of  claim 73 , wherein the yeast cell has been preoptimized for lipid overproduction. 
     
     
         83 . A method for production of a Type III polyketide comprising:
 a. culturing a genetically modified oleaginous yeast cell comprising a gene encoding Acetyl-CoA Carboxylase (AceCarb (ACC1); YALI0C11407g), wherein the oleaginous yeast cell is  Yarrowia lipolytica , and wherein the oleaginous yeast cell further comprises the following modifications: an overexpression of Putative Pyruvate Decarboxylase 2 (PDC2; YALI0D06930), Acetyl-CoA Synthetase 1 (ACS1; YALI0F05962), Acetaldehyde Dehydrogenase 5 (ALD5; YALI0D07942), or peroxin 10 (PEX10; YALI0C01023) in a growth medium; and   b. isolating the Type III polyketide.   
     
     
         84 . The method of  claim 83 , wherein the oleaginous yeast cell further comprises a heterologous gene encoding a heterologous Type III polyketide synthase, further wherein the Type III polyketide synthase is selected from the group consisting of a CAA86219.2 gene from  Gerbera hybrida , AY517486 gene from  Rheum palmatum , JX840717-JX840724 gene from  Pseudomonas fluorescens , B0LDU5 gene from  Rubus idaeus , B1VLQ8 gene from  Streptomyces griseus , SCO1206 from  Streptomyces coelicolor , BAB12102.2 gene from  Humulus lupulus , or a combination thereof. 
     
     
         85 . The method of  claim 83 , wherein the gene is incorporated into a gene expression cassette comprising a promoter and the gene, wherein the gene expression cassette is integrated into a genome of the yeast cell. 
     
     
         86 . The method of  claim 83 , wherein at least two copies of the gene are present in a genome of the yeast cell. 
     
     
         87 . The method of  claim 83 , wherein the gene is episomally expressed from a plasmid. 
     
     
         88 . The method of  claim 83 , wherein the yeast cell comprises an additional genetic modification. 
     
     
         89 . The method of  claim 88 , wherein the additional genetic modification increase the acetyl-CoA or malonyl-CoA levels or fluxes. 
     
     
         90 . The method of  claim 88 , wherein the additional genetic modification increases the rate of beta-oxidation to increase the acetyl-CoA or malonyl-CoA levels or fluxes.