Difference between revisions of "Team:Uppsala/Demonstrate"

Revision as of 01:33, 2 November 2017

<!DOCTYPE html> Results

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.

You can see all of our results on our Results page

Plate with colonies — *E. coli* culture producing colorful pigments from the crocin pathway.

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-       <div style="padding-right:3%; padding-left:3%; text-align:justify;">In our project we have focused on extending the zeaxanthine pathway to produce compounds along the way to crocin. We have stabilized the expression of zeaxanthin by lambda red recombineering and produced zeaxanthin. 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. Crocetin is a biologically interesting and research compound. 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 color change. Our third step was to produce crocin. This is done with enzyme called UGTCs2. Again after creating the biobrick and transforming we have sequenced the plasmid succesfully. We have also transformed the combined BioBrick with CaCCD2, CsADH2946 and UGTCs2 into the zeaxanthin strain.</div>
+       <div style="padding-right:3%; padding-left:3%; text-align:justify;">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 <i>E. coli </i>. 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 <i>E. coli</i> we have sequenced the plasmid succesfully. We have also transformed the combined BioBrick with CaCCD2, CsADH2946 and UGTCs2 into the zeaxanthin strain.</div>
 <div style="padding-right:3%; padding-left:3%; text-align:justify;">You can see all of our results on our <a href= "https://2017.igem.org/Team:Uppsala/Results"> Results page </a></di>