Photoalkanogens with increased productivity
The present disclosure identifies methods and compositions for modifying photoautotrophic organisms as hosts, such that the organisms efficiently convert inorganic carbon and light into n-alkanes, and in particular the use of such organisms for the commercial production of n-alkanes and related molecules.
1. An engineered photosynthetic cell for producing one or more hydrocarbons, wherein said engineered cell comprises (1) one or more recombinant nucleic acid sequences encoding an alkanal deformylative monooxygenase (ADM) enzyme, and (2) a recombinant nucleic acid sequence encoding a formate dehydrogenase enzyme, and wherein upon culture of the engineered photosynthetic cell for at least 2 days the amount of formate accumulation is reduced by at least 5% relative to a control cell that does not comprise a recombinant nucleic acid sequence encoding a formate dehydrogenase enzyme.
2. The engineered cell of claim 1 , wherein said photosynthetic cell is a microbial cell.
3. The engineered cell of claim 2 , wherein said photosynthetic cell is a cyanobacterial cell.
4. The engineered cell of claim 1 , wherein said formate dehydrogenase enzyme is an NAD+-dependent formate dehydrogenase (E.C. 1.2.1.2).
5. The engineered cell of claim 4 , wherein said formate dehydrogenase enzyme is at least 95% identical to SEQ ID NO: 32.
6. The engineered cell of claim 5 , wherein said formate dehydrogenase enzyme is encoded by a nucleotide sequence at least 95% identical to SEQ ID NO: 36.
7. The engineered cell of any of claims 4 - 6 , wherein the expression of said formate dehydrogenase is under the control of a cpcB promoter.
8. The engineered cell of claim 7 , wherein said formate dehydrogenase enzyme is encoded by a plasmid.
9. A method for producing hydrocarbons, comprising:
(i) culturing an engineered photosynthetic cell of claim 1 in a culture medium; and
(ii) exposing said engineered photosynthetic cell to light and inorganic carbon, wherein said exposure results in the conversion of said inorganic carbon by said engineered photosynthetic cell into the hydrocarbons.
10. The method of claim 9 , wherein said photosynthetic cell is a microbial cell.
11. The method of claim 10 , wherein said photosynthetic cell is a cyanobacterial cell.
12. The method of claim 9 , wherein said hydrocarbons comprise n-alkanes which comprise predominantly n-heptadecane, n-pentadecane or a combination thereof.
13. The method of claim 9 , further comprising isolating at least one n-alkane, n-alkene or n-alkanol from said engineered photosynthetic cell or said culture medium.
14. The method of claim 9 , wherein said method reduces formate concentration by at least 5% in said culture comprising said photosynthetic cell.
15. The method of claim 9 , wherein said method reduces formate concentration by at least 35% in said culture comprising said photosynthetic cell.