Microbial production of fatty alcohols
Genes and strains of recombinant microorganisms are provided that are engineered to produce fatty alcohols and fatty alcohol derivatives. The organisms can include one, two, three or more transgenes that direct the biosynthesis of one or more fatty alcohols or derivatives. Methods of producing fatty alcohols using transgenic microorganisms are also provided.
1. A recombinant microorganism, comprising:
at least one exogenous gene encoding an N-terminally truncated eukaryotic alcohol-forming fatty acyl-CoA reductase, wherein the eukaryotic alcohol-forming fatty acyl-CoA reductase is an N-terminally truncated FAR6 of Arabidopsis (SEQ ID NO:28) or a homolog or variant thereof having at least 65% identity to SEQ ID NO:28, or an N-terminally truncated MS2 of Arabidopsis (SEQ ID NO:31) or a homolog or variant thereof having at least 65% identity to SEQ ID NO:31;
wherein the recombinant microorganism produces at least one fatty alcohol.
2. The recombinant microorganism of claim 1 , further comprising at least one exogenous thioesterase gene.
3. The recombinant microorganism of claim 1 , further comprising at least one exogenous acyl-CoA synthetase gene.
4. The microorganism of claim 1 , further comprising at least one exogenous gene encoding a transporter protein.
5. The recombinant microorganism of claim 4 , wherein the at least one exogenous transporter protein gene is CER5, WBC11, a gene encoding AtMRPS, AmiS2, or AtPGP 1, or a gene encoding a homolog or variant of a transporter protein having at least 65% identity to the transporter encoded by the CER5 gene, the transporter encoded by the WBC11 gene, AtMRPS, AmiS2, or AtPGP1.
6. The recombinant microorganism of claim 1 , wherein the microorganism is a photosynthetic microorganism.
7. The recombinant microorganism of claim 6 , wherein the microorganism is a cyanobacterial species.
8. The recombinant microorganism of claim 6 , wherein the microorganism is a eukaryotic microalgal species.
9. A recombinant microorganism comprising:
at least one exogenous gene encoding an N-terminally truncated eukaryotic alcohol-forming fatty acyl-CoA reductase, wherein the N-terminally truncated eukaryotic alcohol-forming fatty acyl-CoA reductase is an N-terminally truncated FAR6 of Arabidopsis (SEQ ID NO:28) or a homolog or variant thereof having at least 65% identity to SEQ ID NO:28, or an N-terminally truncated MS2 of Arabidopsis (SEQ ID NO:31) or a homolog or variant thereof having at least 65% identity to SEQ ID NO:31;
wherein the N-terminal truncation of the eukaryotic alcohol-forming fatty acyl-CoA reductase occurs within ten amino acids of the N-terminus of SEQ ID NO: 28 or SEQ ID NO:31 when the sequence of the eukaryotic alcohol-forming fatty acyl-CoA reductase is aligned for maximum homology with SEQ ID NO:28 or SEQ ID NO:31;
wherein the recombinant microorganism produces at least one fatty alcohol.
10. The recombinant microorganism of claim 5 , wherein the at least one exogenous transporter protein gene is CER5 or a gene encoding a homolog or variant of the CER5 transporter having at least 65% identity to the CER5 transporter.
11. The recombinant microorganism of claim 1 , further comprising at least one exogenous gene encoding an aldehyde-generating fatty acyl-CoA reductase.
12. The recombinant microorganism of claim 11 , wherein the at least one exogenous gene encoding an aldehyde-generating fatty acyl-CoA reductase is an Acr1 gene, an AcrM-1 gene, or one or more of a luxC gene, a luxD gene, or a luxE gene, or a homolog or variant thereof encoding an aldehyde-generating acyl-CoA reductase having at least 65% identity to encoding an aldehyde-generating acyl-CoA reductase encoded by the Acr1 gene, the AcrM-1 gene, a luxC gene, a luxD gene, or a luxE gene.
13. The recombinant microorganism of claim 1 , further comprising at least one exogenous carboxylic acid reductase gene.
14. The recombinant microorganism of claim 7 , wherein the recombinant microorganism is an Agmenellum, Anabaena, Anabaenopsis, Anacystis, Aphanizomenon, Arthrospira, Asterocapsa, Borzia, Calothrix, Chamaesiphon, Chlorogloeopsis, Chroococcidiopsis, Chroococcus, Crinalium, Cyanobacterium, Cyanobium, Cyanocystis, Cyanospira, Cyanothece, Cylindrospermopsis, Cylindrospermum, Dactylococcopsis, Dermocarpella, Fischerella, Fremyella, Geitleria, Geitlerinema, Gloeobacter, Gloeocapsa, Gloeothece, Halospirulina, Iyengariella, Leptolyngbya, Limnothrix, Lyngbya, Microcoleus, Microcystis, Myxosarcina, Nodularia, Nostoc, Nostochopsis, Oscillatoria, Phormidium, Planktothrix, Pleurocapsa, Prochlorococcus, Prochloron, Prochlorothrix, Pseudanabaena, Rivularia, Schizothrix, Scytonema, Spirulina, Stanieria, Starria, Stigonema, Symploca, Synechococcus, Synechocystis, Tolypothrix, Trichodesmium, Tychonema , or Xenococcus species.
15. The recombinant microorganism of claim 1 , wherein the N-terminally truncated eukaryotic alcohol-forming fatty acyl-CoA reductase is an N-terminally truncated FAR6 of Arabidopsis (SEQ ID NO:28) or a homolog or variant thereof having at least 85% identity to SEQ ID NO:28 or an N-terminally truncated MS2 of Arabidopsis (SEQ ID NO:31) or a homolog or variant thereof having at least 85% identity to SEQ ID NO:31.