Figure 5.

Comparison of Tm MiaB activity with components of the TM0927, TM1639, and NADPH reducing system. The activity of Tm MiaB was determined in reactions containing components of the ferredoxin reducing system: (A) ferredoxin TM0927 and NADPH, (B) TM1639 and NADPH, and (C) ferredoxin TM0927, TM1639 and NADPH, and show the formation of ms²i⁶A (black squares), 5′‐dA (red triangles), and SAH (blue circles) at specific time points. Reactions were conducted at 65°C and contained the following in a 100 μL volume: 50 μM Tm MiaB, 150 μM i⁶A tRNA^Phe, 1 mM SAM, 50 μM TM0927, 55 μM TM1639, 1 mM NADPH and buffer system (50 mM HEPES (pH 7.5), 150 mM KCl, 1 mM MgCl₂, and 686 μM L‐(+)‐tryptophan as an internal standard). The reducing system consisting of TM0927 and NADPH (A), TM1639 and NADPH (B), or TM0927, TM1639, and NADPH (C), was incubated for 15 min prior to addition to the reaction containing Tm MiaB.