Difference between revisions of "Team:Uppsala"

This year iGEM Uppsala is Crafting Crocin. Crocin is an apocarotenoid (organic pigment) found in saffron, extracted from Crocus Sativus. Crocin, crocetin, zeaxanthin and other pigments in the crocin pathway, are the compounds which gives saffron it’s beautiful crimson color and delicious taste. The pathway compounds have great potential as organic dyes for industrial applications. On top of this, recent studies suggest that these colorful components have several medicinal properties. The crocin pathway is very poorly characterized and in addition to this the production of saffron is very expensive. Our market analysis show that the harvest of saffron is labour-intensive, but it has a promising increase in market value. By using synthetic biology for recombinant expression in E. coli we hope to reduce the price of the compounds within the crocin pathway and open up the possibility for industrial and medicinal applications to be further explored. To develop our project further, we have done extensive homology modeling, steered molecular dynamics (SMD) and kinetic measurements to characterize the enzymes in our pathway. When working with recombinant microorganisms, and as well when introducing a new manufacturing method to the saffron market, we realized it was very important for us to take ethical aspects into account. To understand the ethical concerns further we spoke to experts, collaborated with other iGEM teams and organized events for the public.

Our achievements this year are:

1. We successfully integrated the first five steps of the crocin pathway from FPP to zeaxanthin into the E. coli chromosome. The result is a E. coli strain expressing zeaxanthin.

2. We have created sequenced verified BioBricks of the three last enzymes in the crocin pathway: CaCCD2, CsADH2946 and UGTCs2. We have also characterized these enzymes with experiments and simulations

3. We are the first to purify and confirm activity of CsADH2946 as well as measuring the kinetic parameters of the enzyme.

4. Finally we have combined the zeaxanthin producing strain with a plasmid containing all three enzymes from zeaxanthin to crocin, thus we successfully constructed a bacterial strain with all the genes needed to produce crocin and the pathway components.