Production of alpha-olefins
The present invention relates to the biosynthesis of α-olefins. In particular, the invention provides methods for the production of medium-chain α-olefins, more particularly C 11 α-olefins, using a polypeptide with decarboxylase activity on free fatty acids with 8 to 14 carbons, in particular on C 8 -C1 2 free fatty acids, more particularly on C 12 free fatty acids, or a genetically engineered host cell expressing or overexpressing said polypeptide.
1. A method for the production of C 7 -C 11 α-olefins comprising culturing a recombinant host cell comprising a recombinant nucleic acid encoding a decarboxylase enzyme comprising the amino acid sequence of SEQ ID NO:13 under conditions suitable for the production of C 7 to C 11 α-olefins by said host cell, wherein the preferred substrate of said decarboxylase enzyme is an C 8 -C 12 free fatty acid.
2. The method of claim 1 , wherein said nucleic acid encoding a decarboxylase enzyme comprises a nucleotide sequence having at least about 75% sequence identity to SEQ ID NO:10 and wherein said recombinant nucleic acid ensures expression or overexpression of said decarboxylase.
3. The method according to claim 1 , wherein C 12 free fatty acids are the preferred substrate of said decarboxylase, and wherein said α-olefins are C 11 α-olefins.
4. The method according to claim 1 , wherein said host cell is cultivated in a medium comprising C 8 -C 12 free fatty acids.
5. The method according to claim 1 , wherein the host cell has further been genetically engineered to produce or overproduce C 8 -C 12 free fatty acids.
6. The method according to claim 5 , wherein the host cell comprises a recombinant nucleic acid encoding an enzyme involved in the production of free fatty acids with a carbon chain length between 8 and 12 free fatty acids.
7. The method according to claim 6 , wherein the host cell comprises a recombinant nucleic acid encoding a thioesterase having activity on C 8 to C 12 acyl-ACP.
8. The method according to claim 1 , wherein the host cell is an oleaginous host cell.
9. The method according to claim 1 , wherein the host cell is selected from the group consisting of bacteria, yeasts, fungi, plants and algae.
10. The method according to claim 1 , further comprising the step of recovering the α-olefins from the host cell or the culture medium.
11. A method for the production of poly-α-olefins comprising the following steps:
i) producing C 7 -C 11 α-olefins according to a method according to claim 1 ; and
ii) performing an oligomerization reaction using the α-olefins obtained in step i) as monomer to produce an oligomer; and optionally,
iii) hydrogenating the oligomer produced in step ii).
12. The method according to claim 11 , wherein the poly-α-olefins are C 33 poly-α-olefins, wherein step i) comprises the production of C 11 α-olefins, and wherein the oligomerization reaction in step ii) is a trimerization reaction.
13. The method of claim 1 , wherein said nucleic acid encoding a decarboxylase enzyme comprises a nucleotide sequence having at least about 80% sequence identity to SEQ ID NO:10 and wherein said recombinant nucleic acid ensures expression or overexpression of said decarboxylase.
14. The method of claim 1 , wherein said nucleic acid encoding a decarboxylase enzyme comprises a nucleotide sequence having at least about 95%, sequence identity to SEQ ID NO:10 and wherein said recombinant nucleic acid ensures expression or overexpression of said decarboxylase.
15. The method according to claim 1 , wherein said host cell is cultivated in a medium comprising C 12 free fatty acids.
16. The method according to claim 1 , wherein the host cell has further been genetically engineered to produce or overproduce C 12 free fatty acids.
17. The method according to claim 5 , wherein the host cell comprises a recombinant nucleic acid encoding an enzyme involved in the production of a C 12 free fatty acid.
18. The method according to claim 6 , wherein the host cell comprises a recombinant nucleic acid encoding a thioesterase having activity on C 12 acyl-ACP.