Methods and compositions for producing alkenes of various chain length
The NonA alkene synthase in Synechococcus sp. displays selective synthesis of 1-nonadecene. Heterologous recombination of a domain, i.e. the acyl binding domain, with other acyl binding proteins, affects acyl substrate chain-length binding selectivity and therefore the chain-length of the synthesized 1-alkenes. Compositions and methods are provided to selectively synthesize 1-alkenes of various chain lengths.
1. An isolated or recombinant chimeric NonA alkene synthase comprising a heterologous acyl binding pocket, wherein said alkene synthase comprises the polypeptide sequence of SEQ ID NO: 30.
2. The alkene synthase of claim 1 , wherein said heterologous acyl binding pocket comprises the polypeptide sequence of SEQ ID NO: 8.
3. The alkene synthase of claim 1 , wherein said alkene synthase is fused to a heterologous amino acid sequence.
4. The alkene synthase of claim 1 , wherein said alkene synthase is encoded by an isolated or recombinant polynucleotide comprising SEQ ID NO: 27.
5. The alkene synthase of claim 1 , wherein said alkene synthase catalyzes the production of an alkene selected from the group consisting of: 1-tridecene, 1-pentadecene, 1-heptadecene, and 1-nonadecene.
6. A method for producing an comprising:
a. culturing a host cell to produce an alkene, wherein said host cell comprises an engineered chimeric NonA comprising a heterologous binding pocket, wherein said alkene synthase comprises the polypeptide sequence of SEQ ID NO: 30; and
b. isolating the alkene.