FIG. 6.

Stereochemical analysis of GGPP enzymatically synthesized by M. smegmatis membranes using ω,E,E-FPP as the allyic primer (B). Assay conditions were as described for Fig. 2. The radiolabeled prenyl diphosphates were dephosphorylated with potato acid phosphatase, and the resulting geranylgeraniol was purified as described for Fig. 5. The radiolabeled geranylgeraniol was applied to a silica gel G60 TLC plate, which was developed in toluene-ethyl acetate (7:3, vol/vol). Radioactive spots were located with a System 200 Imaging Scanner (Bioscan Inc.), and standard polyprenols (A) were located with an anisaldehyde spray reagent (9).