Figure 3.
NNSCAA synthesis by engineered strain WD21 in a 250 mL flask. The cells were grown in 50 mL of LB supplemented with 100 μg/mL ampicillin, 50 μg/mL kanamycin, 0.5 mM of IPTG, 5 g/L l-lysine, 1.0 mM MgSO4, and 0.5 mM ThDP at 37 °C with 250 rpm orbital shaking. Strain WD21 is strain BL21(DE3) plus plasmids pIVC2 and pET21aRKD. The total diamines conclude DAP, HMD, and DAHP. DAP, 1,5-diaminopentane; HMD, 1,6-hexamethylenediamine; and DAHP, 1,7-diaminoheptane. Each experiment was conducted at least three times to ensure reliability and data are presented as mean values ± SD.