Réaction #1244287

ord-c90050b26ee845ae9757a607e0da95a2

Conditions de réaction

Conditions détaillées
See reaction.notes.procedure_details.

Traitement

  1. 1
    AutreBatch fermentations were prepared
  2. 2
    AutreAn overnight
  3. 3
    AutreStar (Invitrogen) was prepared
  4. 4
    workup.ADDITIONAntibiotics were added
  5. 5
    Autrewas grown in an incubator at 37° C. until OD600 reaches about 0.6
  6. 6
    Autrewere grown at 27° C.
  7. 7
    workup.ADDITIONwas added to a final concentration of 0.2 mM
  8. 8
    workup.WAITThe cultures were incubated at 27° C. for 72 hours

Mode opératoire

Batch fermentations were prepared to determine the overall production of flavonoid compounds using commercially available (2S)-eriodictyol. An overnight preinnoculums of recombinant E. coli BL21 Star (Invitrogen) was prepared carrying vectors pCDF-F and pCOLA-DL in LB or M9 minimal media. Antibiotics were added according to manufacturer's instructions (50 ug/mL streptomycin and 30 ug/mL kanamycin). The next day, 4 mL of the preinnoculum was used to seed 76 mL LB or M9 minimal media cultures. The culture was grown in an incubator at 37° C. until OD600 reaches about 0.6. Gene expression was induced by 1 mM ITPG and the cultures were grown at 27° C. with horizontal shaking for 4 hours. (2S)-Eriodictyol dissolved in DMSO was added to a final concentration of 0.2 mM. The cultures were incubated at 27° C. for 72 hours.

Source

DOI: 10.6084/m9.figshare.5104873.v1Brevet: US07807422B2uspto-grants-2010_10