Optimizing Fermentative Production of E. coli

The E. coli strains used in this study are listed in Table 2^{38 (link)}. LB medium was used as the standard medium. For fermentative production, the cells were grown in SM1 minimal medium (1 g/L Trypton, 0.5 g/L Yeast extract, 100 µM potassium phosphate buffer (pH 7.0), 75.7 µM (NH₄)₂SO₄, 1.7 µM NaCl, 1.0 µM MgSO₄, 0.1 µM CaCl₂, 20 mg/L ammonium ferric citrate (AFC), 0.6 µM Na₂MoO₄, 40.4 µM H₃BO₃, 2.9 µM CoCl₂, 1 µM CuSO₄, 8.1 µM MnCl₂, 3.04 mM Met) supplemented with 3% glucose. When required, ampicillin (Ap), tetracycline (Tc), kanamycin (Km) and streptomycin (Sm) were added at concentrations of 50, 10, 50 and 50 µg/ml, respectively. Cultures were incubated aerobically by shaking at 30 °C. Growth was monitored by measurement of the optical density at 562 nm or 660 nm using U-1100 Spectrophotometer (HITACHI).Table 2

E. coli strains used in this study.

Strain	Genotype	Reference or source
BW25113	Wild type, rrnB3 ∆lacZ4787 hsdR514 ∆(araBAD)567 ∆(rhaBAD)568 rph-1	38 (link)
JW3909	BW25113 ∆metJ::Kmr	38 (link)