Part:BBa_K2423005

CaCCD2 with BBa_J04500

This BioBrick codes for the enzyme called CaCCD2 that is the first of three enzymes in the crocin pathway that starts from zeaxanthin. CaCCD2 catalyzes the reaction from zeaxanthin to crocetin dialdehyde. In nature the enzyme can be found in Crocus Sativus (the plant where saffron is harvested from).

This BioBrick contains a lac promoter which means that it has to be induced by lactose or some other closely related derivative, such as IPTG.

Previous iGEM teams have worked with another enzyme called ZCD that was thought to catalyze the same reaction, however no success has been seen with that enzyme.

Usage and Biology

Saffron, a well recognized, but expensive spice has not only uses in terms of cooking but compounds found in saffron have been shown to help with inflammation (1), neurodegenerative diseases (2) and more. Some of those compounds namely zeaxanthin, crocetin dialdehyde, crocetin and crocin are all a part of the same metabolic pathway in the plant specie Crocus Sativus. Not only are these compounds in saffron helpful in terms their medicinal properties, but also the fact that they are very colorful. These aspects was what drew us at iGEM Uppsala 2017 to work with the pathway from zeaxanthin to crocin in the BioBrick format, put also to integrate the metabolic steps that leads up to crocin (the pathway from farnesyl pyrophospate (FPP) to zeaxanthin) on the chromosome of Escherichia Coli. The enzyme presented on this page catalyzes the first reaction in the zeaxanthin to crocin pathway.

In more detail CaCCD2 is an carotenoid cleavage dioxygenase (CCD) that cuts zeaxanthin at 7,8 and 7',8' position forming crocetin dialdehyde and 3-OH-ꞵ-cyclocitral. This subclass enzymes (CCD2) was first mentioned in an article from 2014 (3) and one year later CaCCD2 was cloned and expressed (4).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1939
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 837