Difference between revisions of "Part:BBa K5120001"
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<p>Chalcone Reductase (CHR) is an enzyme that catalyzes a step in the biosynthesis of flavonoids, cconverting 6'-hydroxychalcone (naringenin chalcone) into 6'-deoxychalcone (isoliquiritigenin) by reducing the α,β-unsaturated bond.
 
<p>Chalcone Reductase (CHR) is an enzyme that catalyzes a step in the biosynthesis of flavonoids, cconverting 6'-hydroxychalcone (naringenin chalcone) into 6'-deoxychalcone (isoliquiritigenin) by reducing the α,β-unsaturated bond.
  
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<p>It is constructed for use in plants, specifically <i>Nicotiana benthamiana</i>, to be included in a composite part that allows for the expression of CHS in the biosynthesis of isoflavonoids.</p>
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<p>It is constructed for use in plants, specifically <i>Nicotiana benthamiana</i>, to be included in a composite part that allows for the expression of CHR in the biosynthesis of isoflavonoids.</p>
  
  

Revision as of 03:53, 1 October 2024

Chalcone Synthase (CHS) in Pueraria Mirifica Var. Lobata

Usage and Biology

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Figure 1: Puerarin Pathway

Chalcone Reductase (CHR) is an enzyme that catalyzes a step in the biosynthesis of flavonoids, cconverting 6'-hydroxychalcone (naringenin chalcone) into 6'-deoxychalcone (isoliquiritigenin) by reducing the α,β-unsaturated bond. <p>It is constructed for use in plants, specifically Nicotiana benthamiana, to be included in a composite part that allows for the expression of CHR in the biosynthesis of isoflavonoids.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Proof of Function

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Figure 2: Figure 2. HPLC chromatogram showing the detection of puerarin, daidzin, genistin, iso-vitexin, daidzein and genistein in transformed N. benthamiana samples
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Figure 3: Figure 3, concentration levels of three compounds—daidzein, genistein, and genistin—in Nicotiana benthamiana

This part was used in a composite part along with other key enzymes in the isoflavonoid biosynthetic pathway and was agroinfiltrated into N. benthamiana using Agrobacterium tumefaciens.

After transformation, the modified plants were tested for isoflavonoid production using High-Performance Liquid Chromatography (HPLC). The chromatogram shows the amounts of each target isoflavonoid: puerarin, daidzein, and genistein with the first peak, observed at around 16.0 minutes, representing puerarin, followed by a peak at approximately 17.0 minutes, which corresponds to daidzin. Further along, a peak at 22.0 minutes is attributed to genistin. Traces of all three compounds were detected in N. benthamiana, a plant that does not naturally produce any of these because it lacks the enzymes needed to do so. This shows that the sequence for CHR did function as intended because if it hadn't then the pathway wouldn't have progressed further and produced these isoflavonoids.


Functional Parameters