Difference between revisions of "Part:BBa K3846003"
 
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This enzyme is involved in the biosynthesis of the antimalarial endoperoxide artemisinin, and catalyzes three consecutive oxidations of amorpha-4,11-diene to produce artemisinic acid, with artemisinic alcohol and artemisinic aldehyde as intermediates products. It shows no activity with limonene, alpha-pinene, beta-pinene, pinocarveol, (-)-alloisolongifolene, caryophyllene, (-)-alpha-gurjunene, (+)-gamma-gurjunene, (+)-ledene, (+)-beta-selinene and (+)-valencene as substrates.  
 
This enzyme is involved in the biosynthesis of the antimalarial endoperoxide artemisinin, and catalyzes three consecutive oxidations of amorpha-4,11-diene to produce artemisinic acid, with artemisinic alcohol and artemisinic aldehyde as intermediates products. It shows no activity with limonene, alpha-pinene, beta-pinene, pinocarveol, (-)-alloisolongifolene, caryophyllene, (-)-alpha-gurjunene, (+)-gamma-gurjunene, (+)-ledene, (+)-beta-selinene and (+)-valencene as substrates.  
  
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The related phytobrick part [https://parts.igem.org/Part:BBa_K3846107 BBa_K3846107] was used for further assemblies.  
+
The related phytobrick part [https://parts.igem.org/Part:BBa_K3846107 BBa_K3846107] was used for all assemblies.  
  
  

Latest revision as of 14:09, 19 October 2021


Amorpha-4,11-diene 12-monooxygenase (CYP71AV1)

This part only contains the codon-optimised coding sequence of the Amorpha-4,11-diene 12-monooxygenase (CYP71AV1) from Artemisia annua. This enzyme is involved in the biosynthesis of the antimalarial endoperoxide artemisinin, and catalyzes three consecutive oxidations of amorpha-4,11-diene to produce artemisinic acid, with artemisinic alcohol and artemisinic aldehyde as intermediates products. It shows no activity with limonene, alpha-pinene, beta-pinene, pinocarveol, (-)-alloisolongifolene, caryophyllene, (-)-alpha-gurjunene, (+)-gamma-gurjunene, (+)-ledene, (+)-beta-selinene and (+)-valencene as substrates.

The related phytobrick part BBa_K3846107 was used for all assemblies.


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.