Team:ITB INDONESIA/idea/production-module
RhamCOLIpid
Engineered E. coli producing rhamnolipid for green enhanced oil recovery
Production Module
Part submission tim ITB_Indonesia for production module, Rhamnolipid Source: http://parts.igem.org/Part:BBa_K1685011
There are two enzymes responsible for rhamnolipid production in Pseudomonas aeruginosa. The first enzyme, RhlA, catalyses dimerization of two hydroxyacyl (usually hydroxydecanoyl) chains that will constitute rhamnolipid’s lipid moiety, HAA. RhlB will catalyse the transfer of rhamnose and HAA forming mono-rhamnolipid.
We utilise The Biosurfactor biobrick (BBa_K653000) designed by Panama 2011 that encodes rhlA and rhlB. We put the biobrick under T7lac promoter with strong RBS, and double terminator.
Our chassis for production is E. coli BL21 (DE3). In its genome, it has a T7 coding gene under pLac promoter.
E. coli produces LacI protein to control expression of genes under Lac promoter. LacI will bind into the lac operator, and inhibit the expression of T7.
When lactose is available, it will bind into LacI protein. The binding causes the LacI to dissociate from lac operator.
Without the inhibition from LacI, T7 polymerase will be expressed
The absence of LacI binding to the operator lac in our construct plus the presence of T7 polymerase will allow the expression of our gene which is under T7 promoter. Therefore, rhlAB gene will be expressed to produce rhlA and rhlB enzymes. These enzymes will then produce the rhamnolipid.
