Manipulation of acyl-coa binding protein expression for altered lipid production in microbial hosts
AssigneeE. I. Du Pont de Nemours and Company
InventorsSeung-Pyo Hong, Zhixiong Xue
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Abstract
Acyl-CoA binding protein [“ACBP”] binds thiol esters of long fatty acids and coenzyme A in a one-to-one binding mode with high specificity and affinity. This protein is expected to play an important role in altering lipid production in oleaginous microbial organisms. Knock-out of the protein in the oleaginous yeast, Yarrowia lipolytica, results in a decrease in the total lipid content, while overexpression results in an increase in the total lipid content of the recombinant Yarrowia cells.
Claims19
An oleaginous microbial organism, comprising:
- (i) a recombinant construct comprising at least one isolated polynucleotide comprising a nucleic acid sequence encoding an acyl-CoA binding protein operably linked to at least one regulatory sequence;
- and
- (ii) a source of fatty acids.
An oleaginous microbial organism, comprising:
- (i) a disruption in a native gene encoding an acyl-CoA binding protein;
- and
- (ii) a source of fatty acids.
A method for modifying total lipid content in an oleaginous microbial organism, comprising:
- a) providing an oleaginous microbial organism, comprising:
- (i) a recombinant construct comprising at least one isolated polynucleotide comprising a nucleic acid sequence encoding an acyl-CoA binding protein operably linked to at least one regulatory sequence;
- and
- (ii) a source of fatty acids;
- b) growing the cell of step (a) under conditions whereby transfer of the fatty acids to lipid fractions of the organism is altered by altering expression of the isolated polynucleotide comprising a nucleic acid sequence encoding an acyl-CoA binding protein;
- and
- c) optionally recovering the total lipid fractions of step (b).
A method for modifying fatty acid composition in an oleaginous microbial organism, comprising:
- (a) providing an oleaginous microbial organism, comprising:
- (i) a recombinant construct comprising at least one isolated polynucleotide comprising a nucleic acid sequence encoding an acyl-CoA binding protein operably linked to at least one regulatory sequence;
- and
- (ii) a functional ω-3/ω-6 fatty acid biosynthetic pathway comprising at least one isolated polynucleotide comprising a nucleic acid sequence encoding a desaturase enzyme for biosynthesis of fatty acids;
- (b) growing the cell of step (a) under conditions whereby the expression of the acyl-CoA binding protein is altered, resulting in an altered rate of desaturation by the at least one desaturase enzyme and an altered transfer of the synthesized fatty acids to lipid fractions of the organism;
- (c) optionally recovering the lipid fractions of step (b);
- and,
- (d) optionally determining the modified fatty acid composition of the lipid fractions of step (c).
A method for increasing the total lipid content in a Yarrowia sp., comprising:
- a) providing Yarrowia sp., comprising:
- i) a recombinant construct comprising at least one isolated polynucleotide comprising a nucleic acid sequence encoding an acyl-CoA binding protein, said protein selected from the group consisting of:
- (a) a protein consisting essentially of the sequence set forth in SEQ ID NO:2;
- and
- (b) a protein comprising the following amino acid sequence motifs:
- SEQ ID NO:36, SEQ ID NO:37 and SEQ ID NO:38;
- wherein said isolated polynucleotide is operably linked to at least one regulatory sequence;
- and
- b) a source of fatty acids;
- c) growing the Yarrowia sp. of step (a) under conditions whereby the expression of the acyl-CoA binding protein results in an increased total lipid content of at least 5% when compared to the total lipid content of a Yarrowia sp. lacking said recombinant construct;
- and,
- d) optionally recovering the total lipids of step (b).
Dependencies are inferred from the claim text.