Part:BBa_K2588022

myristic acid synthesis module, glucose-induced

This module consists of the 5 genes encoding the 4 subunits of the actetyl-coenzyme A carboxylase complex and the thioesterase 1 from the e. coli genome. These proteins play a vital role in the fatty acid bio-synthesis in e. coli and lead to a significantly higher production of the C-14 unsaturated fatty acid (myristic acid) as seen in fig. 1.¹ The ribosome binding sites from the registry and their respective strengths used in this assembly were chosen in order to account for the fact that accB and accC are transcribed as a single mRNA and accA and accD encode for subunits alpha and beta of the acetyl-CoA carboxyltransferase, which occurs as an α₂β₂ heterotetramer.^2,3

Fig. 1 (a) fatty acid synthesis in e. coli. (b) fatty acid production. (c) HPLC of fatty acids phenacyl esters.¹

References

1. Xu, P. et al. Modular optimization of multi-gene pathways for fatty acids production in E. coli. Nat. Commun. 4, 1409 (2013).
2. Li, S. J. & Cronan, J. E. The gene encoding the biotin carboxylase subunit of Escherichia coli acetyl-CoA carboxylase. J. Biol. Chem. 267, 855–63 (1992).
3. Broussard, T. C., Price, A. E., Laborde, S. M. & Waldrop, G. L. Complex formation and regulation of escherichia coli acetyl-CoA carboxylase. Biochemistry (2013). doi:10.1021/bi4000707

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 144
  Illegal NheI site found at 167
  Illegal NotI site found at 1841
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Unknown
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Unknown
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 1035
  Illegal SapI.rc site found at 1232