Translational_Unit

Part:BBa_K3846355

Designed by: Marcel Arvid Zimmeck   Group: iGEM21_Hamburg   (2021-10-16)


Costunolide operon

Parthenolide is an interesting sesquiterpene lactone which exhibits a broad spectrum of biological activities (anti-cancer drug and migraine prophylaxis) and can be isolated from Tanacetum parthenium . Parthenolide is derived from the terpene germacrene A, which itself is a cyclization product of FPP by the germacrene A synthase. The biosynthesis of parthenolide occurs through several CYP450 enzymes (germacrene A hydroxylase, costunolide synthase and parthenolide synthase). We organised the necessary enzyme of this pathway also in an operon structure under the control of an IPTG-inducible T7 promoter.

BBa_K3846355

This part describes the costunolide operon containing germacrene A synthase and hydroxylase as well as costunolide synthase leading to the creation of costunolide from FPP. Costunolide as precursor of parthenolide itself has shown to exhibit biological activities. Additionally it is the precursor of various other biological molecules which yet need to be characterised but might prove interesting in later studies.

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


iGEM Hamburg 2021 part collection

Terpenoids are an important group of natural products used as biofuels, drugs or fragrances. Naturally occuring in plants it has been shown that microbial terpene production in microorganisms like yeast, E. coli or cyanobacteria is possible. Nevertheless iGEM projects seem to rarely focus on this interesting class of natural products which is correlated with a lack of useful parts inside the iGEM registry.

Fortunately we were able to change that and designed a novel golden gate based toolbox which allows.

  1. production of terpenoid precursors and simple terpenoids
  2. creation of CYP P450-reductase fusion enzymes to optimise processing of terpenoid precursors and production of bioactive target products
  3. modularity of the system to enable exchange of linker sequences/promoters/etc. (MoClo-compatible toolbox)

MoClo-based Part Design 2.0

To improve the usefulness of our parts, we then aimed to make our parts compatible with the MoClo standard of goten gate based IIS restriction enzyme assembly. Thereby we expanded the Common Genetic Syntax for fusion sites to allow the creation of a) fusion proteins connected by linker sequences and b) multiple CDS expressed in an operon. More useful information and an overview of all our parts can be found on our wiki.

Figure 1: MoClo syntax of the part collection.


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