Enhanced
The invention analyzed a protein sequence using the Udwary-Merski algorithm and identified a tetradomain fragment (DH1-DH2-UMA) which consists of two predicted DH-like domains and two pseudodomains N-terminal to them. This arrangement of domains and pseudodomains is fundamentally the opposite of what is typically observed in the DH cassettes of actinobacterial polyketide synthases or mammalian fatty acid synthases, both of which feature C-terminal pseudodomains. The invention modified E. coli by over expressing DH1-DH2-UMA in E. coli resulting in an increase in the overall production of all the fatty acids normally present in the E. coli fatty acid profile.
1. A gene vector for modifying Escherichia coli ( E. coli ) comprising:
the dehydratase tetradomain gene fragment DH1-DH2-UMA from Photobacterium profundum of SEQ ID:17 cloned into the plasmid vector of SEQ ID:15.
2. The gene vector of claim 1 , wherein said dehydratase tetradomain gene fragment encodes a DH1-DH2 protein.
3. The gene vector of claim 1 , wherein said gene vector modifies E. coli to produce about 4-5 times more free saturated and monounsaturated fatty acids than wild-type E. coli.
4. The gene vector of claim 1 , wherein said dehydratase tetradomain gene fragment is over-expressed at about room temperature.
5. A method for increasing the production of free saturated and monounsaturated fatty acids in E. coli comprising:
cloning the dehydratase tetradomain gene fragment DH1-DH2-UMA from Photobacterium profundum of SEQ ID: 17 into the plasmid vector of SEQ ID:15; and
inserting said cloned vector into E. coli.
6. The method of claim 5 , wherein said dehydratase tetradomain gene fragment is over-expressed at about room temperature.
7. The method of claim 5 , wherein said dehydratase tetradomain gene fragment encodes a DH1-DH2 protein.