Methods and Compositions for the Recombinant Biosynthesis of Terminal Olefins
The present disclosure identifies methods and compositions for modifying microbial cells, such that the organisms efficiently synthesize terminal olefins, and in particular the use of such organisms for the commercial production of propylene and related molecules.
1 . An engineered microbial cell for producing a hydrocarbon, wherein said engineered microbial cell comprises:
a recombinantly expressed protein comprising an engineered sulfotransferase domain at least 90% identical to the sulfotransferase domain of SEQ ID NOs: 4-19; and
a recombinantly expressed protein comprising an engineered thioesterase domain a at least 90% identical to the thioesterase domain of any of SEQ ID NOs: 4-11 and 20-104, wherein said cell synthesizes at least one terminal olefin in amounts greater than that synthesized by an otherwise identical cell lacking said recombinantly expressed activities but cultured under identical conditions.
2 . The engineered microbial cell of claim 1 , wherein said at least one terminal olefin is propylene.
3 . The engineered microbial cell of claim 1 , wherein said engineered microbial cell comprises 3-hydroxybutyryl-ACP.
4 . The engineered microbial cell of claim 1 , wherein said engineered microbial cell comprises a recombinant accBCAD gene or a recombinant fabDHG gene.
5 . The engineered microbial cell of claim 1 , wherein said engineered microbial cell comprises a recombinant 3-hydroxyacyl ACP dehydratase gene, wherein said gene comprises a modification that reduces its expression, comprises a knock-out mutation, or is under the control of an inducible promoter.
6 . The engineered microbial cell of claim 1 , wherein said engineered microbial cell comprises 3-hydroxybutyryl-CoA.
7 . The engineered microbial cell of claim 1 , wherein said engineered microbial cell comprises a recombinant phaA gene or a recombinant phaB gene.
8 . The engineered microbial cell of claim 1 , wherein said at least one terminal olefin is selected from the group consisting of: ethylene, propylene, butylene, butadiene, isoprene, and 1-nonadecene.
9 . The engineered microbial cell of claim 1 , wherein said engineered microbial cell comprises a recombinantly expressed protein comprising any of SEQ ID NOs: 1-3.
10 . The engineered microbial cell of claim 1 , wherein said engineered microbial cell is a cyanobacterium.
11 . A method for producing a terminal olefin, comprising:
a. culturing an engineered microbial cell of claim 1 in a culture medium, wherein said cell synthesizes at least one terminal olefin.
b. isolating said terminal olefin from said microbial cell or said culture medium.
12 . The method of claim 11 , wherein said at least one terminal olefin is propylene
13 . The method of claim 11 , wherein said engineered microbial cell comprises 3-hydroxybutyryl-ACP.
14 . The method of claim 11 , wherein said engineered microbial cell comprises a recombinant accBCAD gene or a recombinant fabDHG gene.
15 . The method of claim 11 , wherein said engineered microbial cell comprises a recombinant 3-hydroxyacyl ACP dehydratase gene, wherein said gene comprises a modification that reduces its expression, comprises a knock-out mutation, or is under the control of an inducible promoter.
16 . The method of claim 11 , wherein said engineered microbial cell comprises 3-hydroxybutyryl-CoA.
17 . The method of claim 11 , wherein said engineered microbial cell comprises a recombinant phaA gene or a recombinant phaB gene.
18 . The method of claim 11 , wherein said at least one terminal olefin is selected from the group consisting of: ethylene, propylene, butylene, butadiene, isoprene, and 1-nonadecene.
19 . The method of claim 11 , wherein said engineered microbial cell comprises a recombinantly expressed protein comprising any of SEQ ID NOs: 1-3.
20 . The method of claim 11 , wherein said engineered microbial cell is a cyanobacterium.