Genetically engineered microbial strains including prototheca lipid pathway genes
Genetically engineered microbial, e.g., Prototheca , cells provide microbial oil useful as a food additive and a source of renewable fuels and industrial chemicals.
1. A recombinant nucleic acid comprising a coding sequence that encodes a Prototheca glycerol-3-phosphate acyltransferase (GPAT) having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 225 and having GPAT activity, wherein said coding sequence is operably linked to a heterologous nucleic acid sequence.
2. The recombinant nucleic acid of claim 1 , wherein said GPAT comprises at least one amino acid substitution in comparison to the amino acid sequence of SEQ ID NO: 225, and has at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 225 and has GPAT activity.
3. The recombinant nucleic acid of claim 1 , wherein said nucleic acid sequence comprises a promoter.
4. The recombinant nucleic acid of claim 1 , wherein said heterologous nucleic acid sequence comprises an untranslated control element.
5. The recombinant nucleic acid of claim 1 , wherein said heterologous nucleic acid sequence encodes a targeting sequence.
6. The recombinant nucleic acid of claim 5 , wherein said targeting sequence is a transit peptide selected from the group consisting of a plastidial targeting sequence and a mitochondrial targeting sequence.
7. The recombinant nucleic acid of claim 1 , wherein said nucleic acid is a DNA molecule.
8. The recombinant nucleic acid of claim 1 , further encoding a sucrose invertase.
9. The recombinant nucleic acid of claim 1 , further encoding an inhibitory RNA that suppresses expression of a Prototheca lipid biosynthesis gene.
10. An expression cassette comprising the recombinant nucleic acid of claim 1 .
11. A genetically engineered microbial cell transformed with the recombinant nucleic acid of claim 1 .
12. A method for obtaining microbial oil comprising culturing the genetically engineered microbial cell of claim 11 under conditions wherein oil is produced, wherein said genetically engineered microbial cell is a genetically engineered Prototheca cell.