Part:BBa_K5120001

Chalcone Reductase (CHR) in Pueraria Mirifica Var. Lobata

Usage and Biology

File:Projectdescription1.png

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.

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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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.