Part:BBa_K1602000

Inducible itaconic acid producing operon

Itaconic acid is an organic, dicarboxylic acid that is biotechnologically synthesized most commonly in Aspergillus terreus. It is derived from citric acid via 2 intermediates and a final decarboxylation.
To enable this pathway in Escherichia coli it is necessary to introduce 3 genes. 2 of which are already established in the citric acid cycle and one that is from Apergillus terreus, a cis-aconitate decarboxylase (cadA).

Figure 1 Reaction scheme of the itaconic acid producing operon. The substrates for the reaction are acetyl-coa and oxaloacetate. To simplify the process we are only looking at oxaloacetate right now. Oxaloacetate is metabolized to itaconic acid in 3 steps.

Usage

This part is a composite of three genes, each with a strong RBS (BBa_B0034) and all of them under control of one T7 Promoter (BBa_K1497017).

citrate synthase - gltA (BBa_K1033001) aconitate hydratase 1 - acnA (BBa_K1033000) cis-aconitate decarboxylase - cadA (BBa_K1497100)
	Figure 2 Genetic map of the itaconic acid producing operon with T7 promoter. This brick directs the flux towards and finally enables E.Coli BL21 cells to synthesize itaconic acid in presence of the inductor IPTG.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  Illegal NheI site found at 1147
  Illegal NheI site found at 4674
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1714
  Illegal BglII site found at 4683
  Illegal XhoI site found at 3681
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1820
  Illegal NgoMIV site found at 2652
  Illegal NgoMIV site found at 4654
  Illegal NgoMIV site found at 5230
  Illegal AgeI site found at 1915
  Illegal AgeI site found at 2081
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 3110
  Illegal BsaI site found at 4603
  Illegal BsaI.rc site found at 1346
  Illegal BsaI.rc site found at 4085