Production of fatty alcohols with fatty alcohol forming acyl-CoA reductases (FAR)
The disclosure relates to methods of producing fatty alcohols from recombinant host cells comprising genes encoding heterologous fatty acyl-CoA reductase (FAR) enzymes. The disclosure further relates to FAR enzymes and functional fragments thereof derived from marine bacterium and particularly marine gamma proteobacterium such as Marinobacter and Oceanobacter ; polynucleotides encoding the FAR enzymes and vectors and host cells comprising the same.
1. A method of producing a fatty alcohol composition comprising:
a) culturing a recombinant microorganism in a suitable culture medium, wherein the recombinant microorganism comprises a gene encoding a heterologous fatty acyl reductase (FAR) enzyme having at least 75% sequence identity to SEQ ID NO: 2, and
b) allowing expression of said gene, wherein said expression results in the production of a composition of fatty alcohols.
2. The method according to claim 1 , wherein the recombinant microorganism is selected from the group of a bacterium, a filamentous fungus, a yeast, and an algae.
3. The method according to claim 2 , wherein the recombinant microorganism is a bacterium.
4. The method according to claim 3 , wherein the recombinant bacterium is an E. coli strain.
5. The method according to claim 1 , wherein the FAR enzyme has at least 85% sequence identity to SEQ ID NO: 2.
6. The method according to claim 4 , wherein the FAR enzyme has at least 85% sequence identity to SEQ ID NO: 2.
7. The method according to claim 1 , further comprising isolating the composition of fatty alcohols.
8. The method according to claim 1 , wherein the produced fatty alcohols include C12-C18 fatty alcohols.
9. The method according to claim 1 , wherein the produced fatty alcohols include a mixture of at least two of C14:0, C16:0, C18:0, C16:1 and C18:1 fatty alcohols.
10. The method according to claim 1 , wherein at least 25% of the produced fatty alcohols are secreted.
11. The method according to claim 10 , wherein at least 50% of the produced fatty alcohols are secreted.
12. The method according to claim 1 , wherein the recombinant microorganism produces at least 0.5 g/L of fatty alcohol.
13. The method according to claim 12 , wherein the recombinant microorganism produces at least 5 g/L of fatty alcohols.
14. The method according to claim 1 , wherein the gene encoding the FAR enzyme comprises a codon optimized nucleic acid sequence.
15. The method according to claim 14 , wherein the nucleic acid sequence is a sequence having at least 70% sequence identity SEQ ID NO:1 or SEQ ID NO:3.
16. The method according to claim 1 , wherein the nucleic acid sequence is integrated into the chromosome of the recombinant microorganism.
17. The method according to claim 1 , wherein the nucleic acid is contained in an expression vector in the recombinant microorganism.
18. The method according to claim 1 , wherein the culturing conditions comprise a temperature in the range of20° C. to 45° C.; a pH in the range of 5 to 7, a time period in the range of 16 hours to 144 hours; and an assimilable carbon source comprising fermentable sugars obtained from a cellulosic feedstock.
19. The method according to claim 1 further comprising reducing the fatty alcohol composition to yield an alkane composition.
20. The method according to claim 1 further comprising esterifying the fatty alcohol composition to produce fatty esters.