Reaktion #1244287

ord-c90050b26ee845ae9757a607e0da95a2

Reaktionsgleichung

NC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O[C@H]3O[C@H](CO)[C@@H](O)[C@H](N)[C@H]3O)[C@H](N)C[C@@H]2N)[C@H](O)[C@@H](O)[C@@H]1O
kanamycin
CN[C@@H]1[C@H](O[C@H]2[C@H](O[C@H]3[C@H](O)[C@@H](O)[C@H](NC(=N)N)[C@@H](O)[C@@H]3NC(=N)N)O[C@@H](C)[C@]2(O)C=O)O[C@@H](CO)[C@H](O)[C@H]1O
streptomycin
CC(C(=N)N)C(=S)NCC(=O)O
ITPG
CS(C)=O
DMSO
→
O=C1C[C@@H](c2ccc(O)c(O)c2)Oc2cc(O)cc(O)c21
Eriodictyol

Edukte

—
flavonoid
NC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O[C@H]3O[C@H](CO)[C@@H](O)[C@H](N)[C@H]3O)[C@H](N)C[C@@H]2N)[C@H](O)[C@@H](O)[C@@H]1O
kanamycin
—
pCDF-F
CN[C@@H]1[C@H](O[C@H]2[C@H](O[C@H]3[C@H](O)[C@@H](O)[C@H](NC(=N)N)[C@@H](O)[C@@H]3NC(=N)N)O[C@@H](C)[C@]2(O)C=O)O[C@@H](CO)[C@H](O)[C@H]1O
streptomycin
CC(C(=N)N)C(=S)NCC(=O)O
ITPG
—
(2S)-eriodictyol
CS(C)=O
DMSO
—
(2S)-Eriodictyol

Reagenzien

Keine

Reaktionsbedingungen

Detaillierte Bedingungen
See reaction.notes.procedure_details.

Aufarbeitung

  1. 1
    SonstigeBatch fermentations were prepared
  2. 2
    SonstigeAn overnight
  3. 3
    SonstigeStar (Invitrogen) was prepared
  4. 4
    workup.ADDITIONAntibiotics were added
  5. 5
    Sonstigewas grown in an incubator at 37° C. until OD600 reaches about 0.6
  6. 6
    Sonstigewere 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

Vorschrift

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.

Quelle

DOI: 10.6084/m9.figshare.5104873.v1Patent: US07807422B2uspto-grants-2010_10