Reaktion #1478586

Reaktionsgleichung

kanamycin

streptomycin

ITPG

DMSO

→

Naringenin

Reaktionsbedingungen

Aufarbeitung

1. 1
   SonstigeBatch fermentations were prepared
2. 2
   SonstigeAn overnight
3. 3
   workup.ADDITIONAntibiotics were added
4. 4
   Sonstigewas grown in an incubator at 37° C. until OD600 reaches about 0.6
5. 5
   Sonstigewere grown at 27° C.
6. 6
   workup.ADDITIONwas added to a final concentration of 0.2 mM
7. 7
   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 (2R/S)-naringenin. An overnight preinnoculums of recombinant E. coli BL21Star (Invitrogen) 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. (2R/S)-Naringenin dissolved in DMSO was added to a final concentration of 0.2 mM. FeSO4 was added to a final concentration of 0.1 mM. The cultures were incubated at 27° C. for 72 hours.

Quelle