Recombinant synthesis of medium chain-length alkanes
The present disclosure identifies methods and compositions for modifying photoautotrophic organisms as hosts, such that the organisms efficiently produce alkanes, and in particular the use of such organisms for the commercial production of alkanes and related molecules. Other materials, methods, and compositions are also described.
1. An engineered cyanobacterium comprising an alkane deformylative monooxygenase encoded by adm, a thioesterase encoded by fatB, fatB2, tesA, FatB3, or Ua-FatB1, and a carboxylic acid reductase encoded by car, carB, or fadD9, wherein the engineered cyanobacterium produces at least one of heptane, nonane, and undecane in an amount greater than an otherwise identical cyanobacterium, cultured under identical conditions, but lacking the alkane deformylative monooxygenase, the thioesterase, and the carboxylic acid reductase.
2. The engineered cyanobacterium of claim 1 , wherein the cyanobacterium further comprises one or more recombinant genes encoding a phosphopanthetheinyl transferase.
3. The engineered cyanobacterium of claim 1 , wherein the cyanobacterium further comprises one or more recombinant genes encoding a phosphopanthetheinyl transferase having EC number 2.7.8.7.
4. The engineered cyanobacterium of claim 3 , wherein the phosphopanthetheinyl transferase is encoded by entD.
5. The engineered cyanobacterium of claim 1 , wherein the cyanobacterium is a thermotolerant cyanobacterium, or a Synechococcus species.
6. The engineered cyanobacterium of claim 1 , wherein expression of an operon comprising one or more of the recombinant genes is controlled by a recombinant promoter, and wherein the promoter is constitutive or inducible.
7. The engineered cyanobacterium of claim 1 , wherein the thioesterase converts C (8-12) acyl-ACP to C (8-12) fatty acid, wherein the carboxylic acid reductase converts C (8-12) fatty acid to C (8-12) aldehyde, and wherein the alkane deformylative monooxygenase converts C (8-12) aldehyde to C (7-11) alkane.
8. A cell culture comprising a culture medium and the cyanobacterium of claim 1 .
9. A method for producing hydrocarbons, comprising:
culturing an engineered cyanobacterium of claim 1 in a culture medium, wherein the engineered cyanobacterium produces increased amounts of alkanes relative to an otherwise identical cyanobacterium, cultured under identical conditions, but lacking the recombinant genes.
10. A method for producing hydrocarbons, comprising:
(i) culturing an engineered cyanobacterium of claim 1 in a culture medium; and
(ii) exposing the engineered cyanobacterium to light and inorganic carbon, wherein said exposure results in the conversion of said inorganic carbon by the cyanobacterium into alkanes, wherein the alkanes are produced in an amount greater than that produced by an otherwise identical cyanobacterium, cultured under identical conditions, but lacking the recombinant genes.
11. The engineered cyanobacterium of claim 1 , wherein the alkane deformylative monooxygenase has EC number 4.1.99.5.
12. The engineered cyanobacterium of claim 1 , wherein the thioesterase has EC number 3.1.2.14.
13. The engineered cyanobacterium of claim 1 , wherein the carboxylic acid reductase has EC number 1.2.99.6.
14. The engineered cyanobacterium of claim 1 , wherein the cyanobacterium is a Synechococcus species.
15. The engineered cyanobacterium of claim 1 , wherein the thioesterase is encoded by fatB, fatB2, or tesA.
16. The engineered cyanobacterium of claim 1 , wherein the thioesterase is encoded by fatB.
17. The engineered cyanobacterium of claim 1 , wherein the thioesterase is encoded by fatB2.
18. The engineered cyanobacterium of claim 1 , wherein the thioesterase is encoded by tesA.
19. The engineered cyanobacterium of claim 1 , wherein the carboxylic acid reductase is encoded by carB.