Part:BBa_K1702000

Overexpression of E. coli fadD under control of lac promoter

Acyl-CoA synthetase is the first enzyme the beta oxidation pathway. It attaches Coenzyme-A to long chain fatty acids (LCFAs), allowing them to continue through the pathway. The transcription of fadD is controlled by the enzyme produced by fadR under standard conditions. In this plasmid, fadD is under control of the lac promoter.
While there is not a published structure of fadD in E. coli Andersson, CS et al. published a structure from Mycobacterium tuberculosis.



Experimental Characterization
The intended use of this part was to increase beta-oxidation, which is the natural process for the breakdown of fatty acids, in order to allow for frying oil waste to be used as an alternative substrate. We ran several experiments on this part and found that it did not significantly improve E. coli's ability to grow on fatty acids or frying oil. For exact experimental protocol see our wiki. Our first experiment was on 0.04% steric acid (a mimic for used frying oil components) and we saw significantly better growth from our control (just the lac promoter) than we did from our construct (lac promoter + fadD induced with IPTG).

In an experiment with cells grown on 0.04% frying oil we did not see significant improvement by the fadD gene as compared to the control. This experiment got slightly odd data and can not be used to definitively say what was occurring.



Overall this part by itself does not greatly improve the growth of E coli. on frying oil.

Sequence and Features

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
  COMPATIBLE WITH RFC[1000]