Team:Uppsala/Demonstrate
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Demonstrate
In our project we have focused on producing the colorful and valuable molecules from the crocin pathway, naturally found in saffron. We have stabilized the expression of the saffron component zeaxanthin by lambda red recombineering and produced zeaxanthin in E. coli . The next goal was to introduce further steps in the pathway. In the first step we included the enzyme CaCDD2 with a lac promotor. The enzyme construct was verified by successful sequencing. Then we introduced another step: CsADH2946, the enzyme that leads to Crocetin. We have successfully expressed and purified this enzyme and tested its activity. Both CaCDD2 and CsADH2946 were transformed into the zeaxanthin strain and production of crocetin was observed by a change in color. The next step was to produce crocin from crocetin, a reaction catalyzed by the enzyme UGTCs2. Again after creating the biobrick and transforming it into E. coli we have sequenced the plasmid succesfully. We have also transformed the combined BioBrick with CaCCD2, CsADH2946 and UGTCs2 into the zeaxanthin strain.
Detailed results that show that our project works can be found on our Results page
