In Vitro Optimization of Enzymes Involved in Precorrin-2 Synthesis Using Response Surface Methodology
Fig 4
Titration of each enzyme involved in precorrin-2 synthesis.
The reaction mixture contained: 5 mM ALA, 200 μM SAM, 1 mM NAD, 1 μM precorrin-2 dehydrogenase, and various concentrations of titrated enzymes. (A), Optimization of PBGS concentration. The reaction mixture contained 1 μM PBGD, 1 μM UROS, 1 μM SUMT, and various concentrations of PBGS from 0.02–3 μM. (B), Optimization of PBGD concentration. The reaction mixture contained 0.1 μM PBGS, 1 μM UROS, 1 μM SUMT, and various concentrations of PBGD from 0.1–6 μM. (C), Optimization of UROS concentration. The reaction mixture contained 0.1 μM PBGS, 1 μM PBGD, 1 μM SUMT, and various concentrations of UROS from 0.1–10 μM. (D), Optimization of SUMT concentration. The reaction mixture contained 0.1 μM PBGS, 1 μM PBGD, 1 μM UROS, and various concentrations of SUMT from 0.1–35 μM. Results are presented as the mean of 3 replicates. Error bars indicate SD.