Difference between revisions of "Part:BBa K1497018"

 
 
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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.
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The dihydroflavonol 4-reductase (DFR; EC 1.1.1.219) from the plant <i> Dianthus caryophyllus </i> 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<sup>+</sup> instead of NADP<sup>+</sup> (Queen Mary University of London 2014). DFR contains 353 amino acids and has a molecular weight of approximately 39.4 kDa. <br> <br>
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The iGEM Team TU Darmstadt 2014 used the DFR in combination with the <a href="/Part:BBa_K1497023">B0034</a>-RBS  and verified the function of the DFR in their pelargonidin operon <a href="/Part:BBa_K1497023">(K1497023)</a>.
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      <br>
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      <p class="MsoCaption" align="text-align:justify"><span lang="EN-US"><b>Figure 1</b></span></a><span lang="EN-US">
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Reaction of the DFR. One Dihydroflavonol reacts with NADPH to form a leucoanthocyanidin. The reverse reaction works with NAD<sup>+</sup> or NADP<sup>+</sup></span></p>
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      <p class="MsoCaption" align="text-align:justify"><span lang="EN-US"><b>Figure 2</b></span></a><span lang="EN-US">
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Reaction of the F3H </span></p>
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<partinfo>BBa_K1497018 parameters</partinfo>
 
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===References===
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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
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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.
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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.
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4. Tanaka Y, Brugliera F, B PTRS, et al. (2013) Flower colour and cytochromes P450 Flower colour and cytochromes P450.

Latest revision as of 01:43, 18 October 2014

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.