Part:BBa_K5136232

I0500-B0034-rhlA-B0034-rhlB-B0015
Responsible for bioproduction of rhamnolipid.

Biology

RhlA, RhlB and RhlC, which are important enzymes to control the production of rhamnolipids, are found almost exclusively in Pseudomonas sp. and Burkholderia sp. (1)

Usage and Design

RhlA is used for generating the lipid component of rhamnolipid, RhlB can then catalyze the reaction between HAA and dTDP-l-rhamnose to form mono-rhamnolipid. We ligased two basic parts, then they are co-expression by the constant promoter BBa_J23100. And the RBS BBa_B0034 as well as the terminator BBa_B0015 together into the expression vector pSB1C3 by standard assembly, then transferred it into E. coli BL21(DE3) to verify its ability to produce rhamnolipids.

Characterization

1. Double transformation

We used double transformation to prove the monorhamnoolipid production. Plasmid BBa_K5136233_pSB3K3 and plasmid BBa_K5136232_pSB1C3 were transformed into E. coli BL21(DE3). The positive transformants were selected by kanamycin and chloramphenicol.

Reference

1. Chong, H.; Li, Q., Microbial production of rhamnolipids: opportunities, challenges and strategies. Microb Cell Fact 2017, 16 (1), 137.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1205
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1144
  Illegal BamHI site found at 1305
  Illegal XhoI site found at 2041
  Illegal XhoI site found at 3288
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 2181
  Illegal NgoMIV site found at 3015
  Illegal AgeI site found at 979
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1530
  Illegal BsaI site found at 2531
  Illegal BsaI.rc site found at 1714
  Illegal SapI site found at 961