Reaktionsgleichung
[CH3:1][C@@H:2]1O[C@@H:5]([O:7][C@H:8]2[C@H:13](O)[C@@H:12]([OH:15])[C@H:11](NC(N)=N)[C@@H:10]([OH:20])[C@@H:9]2NC(N)=N)[C@H:4](O[C@@H]2O[C@@H](CO)[C@H](O)[C@@H](O)[C@@H]2NC)[C@@:3]1(O)C=O.C1[C@H](N)[C@@H:45]([O:48][C@H]2O[C@H](CN)[C@@H](O)[C@H](O)[C@H]2O)[C@H:44](O)[C@@H](O[C@H]2O[C@H](CO)[C@@H](O)[C@H](N)[C@H]2O)[C@@H]1N.[CH3:74][CH:75](C(NCC(O)=O)=S)C(N)=N.CS(C)=[O:88]>>[CH:74]1[C:4]([C@H:5]2[O:7][C:8]3[CH:9]=[C:10]([OH:20])[CH:11]=[C:12]([OH:15])[C:13]=3[C:45](=[O:48])[CH2:44]2)=[CH:3][CH:2]=[C:1]([OH:88])[CH:75]=1Komponenten
Versuchsvorschrift
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.
Aufarbeitung
- customBatch fermentations were prepared
- customAn overnight
- additionAntibiotics were added
- customwas grown in an incubator at 37° C. until OD600 reaches about 0.6
- customwere grown at 27° C.
- additionwas added to a final concentration of 0.2 mM
- waitThe cultures were incubated at 27° C. for 72 hours
Quellen und Nachweise
- ORD-Eintrag
- ord-1454c3e51d1648389a94fa031c335a8d
- Quelldatensatz
- uspto-grants-2008_03
- Patentnummer
- US07338791B2
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