Part:BBa_K1497018

B0034-F3H-B0034-DFR

This part encodes the sequence for the enzyme Flavanone-3ß-hydroxylase (F3H) EC 1.2.7.3 from Petroselinum crispum. F3H catalyzes naringenin, an anthocyanidin precursor, to the substance dihydroflavonol (Tanaka, 2006) and the dihydroflavonol 4-reductase. 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 leucoanthocyanidins. 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 DFR in combination with the B0034-RBS and verified the function of the 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+

Figure 2 Reaction of the F3H

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 185
  Illegal BamHI site found at 675
  Illegal BamHI site found at 1481
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1233
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1218
  

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.

4. Tanaka Y, Brugliera F, B PTRS, et al. (2013) Flower colour and cytochromes P450 Flower colour and cytochromes P450.