Part:BBa_K849003:Design

bifunctional Cyclase from Gibberella fujikuroi

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 373
  

Design Notes

These description is conform to the proposed standardized part description by TU Munich.

Reaction Geranylgeranyl pyrophosphat(GGPP) -> ent-Kaurene

This enzyme is a bifunctional diterpene cyclase which catalyses the reaction from GGPP to gibberellin precursor ent-Kaurene. In many organisms this is done via the intermediate copalyl pyrophosphate. By using this enzyme you can skip this step. The iGEM Team Frankfurt intended to use this biobrick for synthesis of the sweetener Rebaudiosid A from stevia plant in yeast. It is the first step not naturally occuring in yeast.

Related BioBrick:

• Related BioBricks:
  • BBa_K849004: ent-Kaurene Oxidase from Gibberella fujikuroi uses ent-kaurene, the product of this enzyme, to synthesize ent-Kaurenoic acid.

Quality control measures:
Part was totally sequenced

Backbone:

• Backbone name: pSB1C3

Safety notes:

Source:

• Synthesized by life technology.

Organism:

• Genesequence derived from fungus Giberella fujikuroi
• Statement about functionality in other chassis.
  • This part needs geranylgeranyl pyrophosphate as a substrate. This is a product of the mevalonate- and non-mevalonate pathway. Saccharomyces cerevisiae uses the mevalonate pathway and produces GGPP but concentration is very low so overexpression of the mevalonate pathway is necessary. E.coli uses the non-mevalonate pathway.

References

Literature references:

• [http://www.ncbi.nlm.nih.gov/pubmed/9745028 Pubmed: Tudzynski B, Kawaide H, Kamiya Y., 1998: Gibberellin biosynthesis in Gibberella fujikuroi: cloning and characterization of the copalyl diphosphate synthase gene]

Database references:

• [http://www.uniprot.org/uniprot/Q9UVY5 Uniprot Q9UVY5]