Difference between revisions of "Part:BBa K1497013"
 
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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 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. <br> <br>
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The biobrick B0034-DFR is a composite consisting of the biobrick DFR (BBa_K1497013) and RBS (Ellowitz 1999, <a href="/Part:BBa_K1497023">BBa_B0034</a>).
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The iGEM Team TU Darmstadt 2014 used the DFR and verified the function of the DFR in their pelargonidin operon <a href="/Part:BBa_K1497023">(K1497023)</a>.  
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B0034 is a strong ribosome binding site with a biology efficiency of 1.0.<br>
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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 leucoanthocyanidin. 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>
 +
The iGEM Team TU Darmstadt 2014 used the B0034-DFR and verified the function of the B0034-DFR in their pelargonidin operon <a href="/Part:BBa_K1497023">(K1497023)</a>.  
 
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  src="https://static.igem.org/mediawiki/parts/a/ac/DFR_Wiki_reaction.png"></p>
 
  src="https://static.igem.org/mediawiki/parts/a/ac/DFR_Wiki_reaction.png"></p>
 
       <p class="MsoCaption" align="text-align:justify"><span lang="EN-US"><b>Figure 1</b></span></a><span lang="EN-US">
 
       <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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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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====References====
 
====References====
  
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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)<br><br>
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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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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)
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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.

Latest revision as of 01:03, 18 October 2014

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.