Fig. 3
Effect of bacterial cell lysate on the GSH content analysis in LigF reaction system. a Absorbance value OD405; b 96-Well Microplate. 1. The bacterial cell lysate: BL21(DE3) with ligF plasmid; 2. without bacterial cell lysate; 3. the bacterial cell lysate: BL21(DE3) without plasmid. In this part, a 96-Well Microplate was used to investigate the effect of cell lysate on assay of LigF activity by GSH analysis. In No. 1 group, the reaction mixture was 200 μL in total, including 4 μL 20 mM GVG, 4 μL 3 mg/mL GSH, 20 μL cell lysate from BL21(DE3) with ligF plasmid, 172 μL 50 mM Tris–Cl buffer (pH 8.0); in the No. 2 group, 20 μL cell lysate was replaced by the 20 μL 50 mM Tris–Cl buffer (pH 8.0); in the No. 3 group, the cell lysate was from BL21(DE3) strain without plasmid. The mixture was cultured at 27 °C,80 rpm for 30 min. And then, 50-μL sample was taken for the GSH content analysis to describe the LigF activity