Part:BBa_K2819201

Blue light-repressible production of FNS

This part contains the coding sequence of flavone synthase (FNS), under an EL222 blue light repressible promoter, BBa_K2332019. This gene comes from Gerbera hybrida, and the sequence was obtained after codon optimization.

Usage and Biology

- This engineered part can be used to produce Eriodictyol, a flavonoid, in dark conditions.
- We recommend BL21* (DE3) as a chassis for this composite part. This is because BL21* (DE3) is an RNase knockout strain, therefore the half-life of mRNA is prolonged in this strain, which can help in the biomanufacturing of the E. coli.

Characterization

Biosynthesis of Luteolin
FNS is involved in the luteolin-synthesis pathway. We use co-transformed this part with BBa_K2819206 into BL21* (DE3) for the biosynthesis of luteolin. We not only carried out the biosynthesis in flasks, but also in a bioreactor.

Figure 2: Luteolin in the supernatant of the cell cultures from biosynthesis using a bioreactor

High Performance Liquid Chromatography (HPLC)

To confirm that we have produced luteolin, we carried out HPLC, and the following shows the results.

Figure 3: Comparison of Luteolin concentration in BL21* (DE3) wild type and BL21* (DE3) with our parts

Conclusion

As both F3'H and FNS are needed in the biosynthesis of luteolin, these characterisation results indicate that the FNS gene is being produced, however, further optimization is needed to better understand the gene.

Sequence and Features

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 803
  
• 1000
  COMPATIBLE WITH RFC[1000]