Methods and compositions for the recombinant biosynthesis of
The present disclosure identifies methods and compositions for modifying photoautotrophic organisms as hosts, such that the organisms efficiently convert carbon dioxide and light into n-alkanes, and in particular the use of such organisms for the commercial production of n-alkanes and related molecules.
1. A method for producing hydrocarbons, comprising:
(i) culturing an engineered cyanobacterium in a culture medium, wherein said engineered cyanobacterium comprises a recombinant acyl ACP reductase (AAR) enzyme and a recombinant alkanal decarboxylative monooxygenase (ADM) enzyme; and
(ii) exposing said engineered cyanobacterium to light and carbon dioxide, wherein said exposure results in the conversion of said carbon dioxide by said engineered cynanobacterium into n-alkanes, wherein at least one of said n-alkanes is selected from the group consisting of n-tridecane, n-tetradecane, n-pentadecane, n-hexadecane, and n-heptadecane, and wherein the amount of said n-alkanes produced is between 0.1% and 5% dry cell weight and at least two times the amount produced by an otherwise identical cyanobacterium, cultured under identical conditions, but lacking said recombinant AAR and ADM enzymes.
2. The method of claim 1 , wherein at least one of said recombinant enzymes is heterologous with respect to said engineered cyanobacterium.
3. The method of claim 1 , wherein said engineered cyanobacterium further produces at least one n-alkene or n-alkanol.
4. The method of claim 3 , wherein said engineered cyanobacterium produces at least one n-alkene or n-alkanol selected from the group consisting of n-pentadecene, n-heptadecene, and 1-octadecanol.
5. The method of claim 3 , wherein said n-alkanes comprise predominantly n-heptadecane, n-pentadecane or a combination thereof.
6. The method of claim 3 , further comprising isolating at least one n-alkane, n-alkene or n-alkanol from said engineered cyanobacterium or said culture medium.
7. The method of claim 1 , wherein said enzymes are encoded by a plasmid.
8. The method of claim 1 wherein said enzymes are encoded by recombinant genes incorporated into the genome of said engineered cyanobacterium.
9. The method of claim 1 wherein said enzymes are encoded by genes which are present in multiple copies in said engineered cyanobacterium.
10. The method of claim 1 wherein said enzymes are encoded by genes which are part of an operon, and wherein the expression of said genes is controlled by a single promoter.