Design
We aimed to create E. coli that synthesizes palmitoleic acid. The K12 strain of E. coli (E. coli K-12 MG1655) has an enzyme that synthesizes Hexa-decanoyl-[acp], shown in red in the figure.
Since two enzymes (1.14.192 and 3.1.2.14) are required to synthesize hexadec-9-enoic acid, these two genes were targeted in our project, shown in blue in the figure.
At first, we ordered two genes to be synthesized from IDT. (Figure below) shows 1.14.192 as enzyme ① and 3.1.2.14 as enzyme ②.
We uses BBa_J04450 as our vector.
And we introduced genes ① and ② synthesized by IDT into
BBa_J04450.
(BBa_K3303003, BBa_K3303004)
Next, in order to express the gene, BBa_K3303003 containing the
enzyme ① is introduced into BBa_I0500, BBa_I13453, BBa_I14032
containing the promoter.
(BBa_K3303005, BBa_K3303001, BBa_K3303002)
Then the vector containing the promoter and enzyme ① (BBa_K3303005, BBa_K3303001, BBa_K3303002) and the vector containing enzyme ② (BBa_K3303004) are combined and transformed into E. coli.
The fatty acid synthesis is then evaluated using gas chromatography.