Part:BBa_K1497013

B0034-DFR

The biobrick B0034-DFR is a composite consisting of the biobrick DFR (BBa_K1497013) and RBS (Ellowitz 1999, BBa_B0034). B0034 is a strong ribosome binding site with a biology efficiency of 1.0. The dihydroflavonol 4-reductase (DFR; EC 1.1.1.219) from the plant Dianthus caryophyllus is an enzyme, catalyzing the reversible conversion of dihydroflavonols e.g. dihydro-kaempferol or dihydroquercetin into their corresponding leucoanthocyanidin. This reaction is NADPH-dependent (Liew et al. 1998) but its counter reaction can also occur with NAD+ instead of NADP+ (Queen Mary University of London 2014). DFR contains 353 amino acids and has a molecular weight of approximately 39.4 kDa. The iGEM Team TU Darmstadt 2014 used the B0034-DFR and verified the function of the B0034-DFR in their pelargonidin operon (K1497023).

Figure 1 Reaction of the DFR. One Dihydroflavonol reacts with NADPH to form a Leucoanthocyanidin. The reverse reaction works with NAD+ or NADP+.

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 348
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 100
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 85
  

References

Chye-Fong Liew, Chiang-Shiomg Loh, Chong-Jin Goh, Saw-Hoon Lim 1998, 'The isolation, molecular characterization and expression of dihydroflavonol 4-reductase cDNA in the orchid, Bromheadia finlaysoniana', Plant Science,  vol. 135, no. 2, pp. 161-169. Available from: ScienceDirect (31.08.2014)

Queen Mary University of London, EC 1.1.1.219. Available from: <http://www.chem.qmul.ac.uk/iubmb/enzyme/EC1/1/1/219.html> (31.08.2014)