FIG. 5.

Effect of increasing substrate concentration on product chain length. TLC analysis of the radiolabeled products synthesized by Rv2361c in the presence of 30 μM [1-¹⁴C]IPP and 50 μM ω,E,Z-FPP (A) or 200 μM ω,E Z-FPP (B). The alcohols obtained by enzymatic dephosphorylation of the products formed were analyzed by reverse-phase TLC as described in Materials and Methods. Products labeled with [1-¹⁴C]IPP were visualized by a Bioscan System 200 imaging Scanner (Bioscan Inc). Standard polyprenols were located with an anisaldehyde spray reagent. The large peak that goes off the scale in both panels corresponds to the allylic substrate, E,Z-FPP.