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

1. Petit P, Granier T, d’Estaintot BL, et al. (2007) Crystal structure of grape dihydroflavonol 4-reductase, a key enzyme in flavonoid biosynthesis. Journal of molecular biology 368:1345–57. doi: 10.1016/j.jmb.2007.02.088

2. Gollop R, Even S, Colova-tsolova V, et al. (2002) Expression of the grape dihydroflavonol reductase gene and analysis of its promoter region 1. 53:1397–1409.

3. Liew C, Loh C, Goh C, Lim S (1998) The isolation , molecular characterization and expression of dihydroflavonol 4-reductase cDNA in the orchid , Bromheadia. 135:161–169.