Part:BBa_K1685011

Rhamnolipid production

This part encodes two enzymes, rhlA and rhlB (BBa_K653000) designed by Panama 2011, for biosynthesis of rhamnolipid. The enzymes are under T7 promoter with medium strengh and a strong ribosomal binding site (BBa_K613010), and stopped with a double terminator (BBa_B0015).

The promoter had the second highest efficiency as shown by EPFL team 2011 (https://parts.igem.org/Part:BBa_K613010:Experience).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 225
  Illegal BamHI site found at 785
  Illegal XhoI site found at 961
  Illegal XhoI site found at 2247
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1140
  Illegal NgoMIV site found at 1861
  Illegal NgoMIV site found at 1974
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 450
  Illegal BsaI site found at 1490
  Illegal BsaI.rc site found at 634
  

Usage and Biology

The biosynthesis of rhamnolipid is relatively simple. It only needs two enzymes which catalyse sequential reaction. The first enzyme, rhamnosyltransferase A (rhlA), which catalyse the dimerisation of two b-hydroxyacyl-ACP chains to make 3-(3-hydroxyalkanoyloxy)alkanoic acids (HAAs). The HAA will form the lipid moiety of the rhamnolipid. Rhamnosyltransferase B, rhlB, will then join TDP-l-rhamnose with HAA to produce mono-rhamnolipid.

Because Escherichia coli can synthesize both rhamnose and fatty acid, the addition of these two enzymes will be sufficient to give the bacteria the ability to synthesize rhamnolipid.

References

Abdel-Mawgoud et al. 2011. Rhamnolipids: Detection, Analysis, Biosynthesis, Genetic Regulation, and Bioengineering of Production. in G. Soberon‐Chavez (ed.), Biosurfactants, Microbiology Monographs 20. Springer-Verlag Berlin, Heidelberg. pp. 13-55