Part:BBa_K2588015

Leu Operon coding sequences from E. coli

We designed leuABCD to make E. coli produce 3-Methyl-1-butanole as a lure molecule to attract mosquitos. leuABCD is only one part of this lure module. leuABCD consists of four genes leuA, leuB, leuC and leuD. Its natural function is to catalyze the reaction of 2-ketoisovalerate to 2-ketoisocaproate. 2-ketoisocaproate can then react to 3-methyl-1-butanole.

Usage and Biology

leuABCD is a gene naturally occurring in the genome of E. coli. Normally its translation leads to four different proteins which are involved in the synthesis of L-Leucine. LeuA converts 2-ketoisovalerate into 2-isopropylmalate. LeuB and LeuC then convert it into 3-isopropylmalate. LeuD converts it into 2-isopropyl-3-oxosuccinate and 2-isopropyl-3-oxosuccinate then spontaneously transforms into 2-ketoisocaproate. Naturally, it then gets transformed into L-Leucine. That synthesis can be interrupted by using two other genes (kivD from Lactococcus lactis and ADH2 from Saccharomyces cerevisiae) to go from 2-ketoisocaproate to 3-methyl-1-butanol instead of L-Leucine.

LeuABCD is part of the 3MB module, BBa_K2588024

References

1. Connor, M.R., Cann, A.F. & Liao, J.C. Appl Microbiol Biotechnol (2010) 86: 1155.
2. Engineering of an Escherichia coli Strain for the Production of 3-Methyl-1-Butanol, Michael R. Connor, James C. Liao, Appl. Environ. Microbiol. Sep 2008

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 4514
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 3252
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 4340
  Illegal SapI.rc site found at 1172