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Assembling the enzyme gene Rhamnosiltransferase 1 (rhlAB) into a Biobrick-friendly for rhamnolipid production in E. coli |
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PROJECT DESCRIPTION
Rhamnolipids are naturally occurring glycolipids produced by the Pseudomonas aeruginosa. Rhamnolipids have many applications, because of their natural biosurfactant, emulsifying, fungicidal, antibiotic, and pharmaceutical properties. Also rhamnolipids can operate in extreme conditions such as high temperature, pH, and salinity. They have a low toxicity, and they are biodegradable compared to their synthetic chemical counterparts. Rhamnolipid is composed of rhamnose sugar molecule and b-hydroxyalkanoic acid. The rhamnosyltransferase 1 complex (RhlAB) is the key enzyme responsible for transferring the rhamnose moiety to the b-hydroxyalkanoic acid moiety to biosynthesize rhamnolipid. Since the organisms known to produce rhamnolipids are almost exclusively pathogens. For this reason, we propose the non-pathogenic nature of E. coli as an attractive candidate for commercial rhamnolipid production by assembling our rhlAB gene into a BioBrick using synthetic biology to engineer these new rhlAB producing bacteria for human benefit.
Our aim is to construct a BioBrick that leads to the production of rhlAB enzyme complex in E.coli. To accomplish this we must modify the natural gene sequence of rhlAB complex because it includes illegal restriction sites and requires modifications to be fit into a BioBrick. We will perform silent mutations in the gene sequence with a form of mutagenesis. To measure our success we will do bio-assays with our new E. coli rhlAB strain to degrade rhamnose sugars and fatty-acids substrates and turn it in rhamnolipids. We hope that our BioBricks will become a useful tool for the bioengineered industry and widely used as a simple and effective means of rhamnolipid production.
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