Substrate specificities of farnesyl diphosphate synthases with respect to cyclic substrate homologs

Abstract

We investigated substrate specificities of farnesyl diphosphate synthases (FPSs) derived from porcine liver and Bacillus stearothermophilus by examining the reactivity of cyclopentylideneethyl diphosphate with several 3-alkyl homologs of isopentenyl diphosphate. Reaction of cyclopentylideneethyl diphosphate with isopentenyl diphosphate using porcine liver or bacterial enzyme gave 10-cyclopentyliden-3,7-dimethyldeca-2,6-dinenyl diphosphate as a double condensation product, with relative yields of 40.9% for the porcine liver enzyme and 15.9% for the bacterial enzyme. Reaction of cyclohexlideneethyl diphosphate with 3-ethylbut-3-enyl diphosphate using the bacterial enzyme gave 10-cyclohexliden-3,7-diethyldeca-2,6-dinenyl diphosphate (yield: 24.6%).

Substrate Specificities of <i>E</i>-and <i>Z</i>-Farnesyl Diphosphate Synthases with Artificial Substrates

“…3-Ethyl-, 3-propyl-, and 3-butylIPP were synthesized according to our previously reported method [22][23][24][25]. Briefly, 3-ethylIPP was synthesized as follows: 2-(3-chloromethylbut-3-enoyloxy)-tetrahydro-2H-pyran was obtained by selenium oxidation and chlorination of isopentenyl tetrahydropyranyl ether, which was methylated by a Girgnard reaction, yielding 3-ethylbut-3-en-1-ol by deprotection [25].…”

“…Briefly, 3-ethylIPP was synthesized as follows: 2-(3-chloromethylbut-3-enoyloxy)-tetrahydro-2H-pyran was obtained by selenium oxidation and chlorination of isopentenyl tetrahydropyranyl ether, which was methylated by a Girgnard reaction, yielding 3-ethylbut-3-en-1-ol by deprotection [25]. Diphosphorylation of the corresponding alcohols was then carried out using the method of Davisson et al [26].…”

“…Wild-type FPP synthase derived from B. stearothermophilus was purified by following a previously reported method [20][21][22][23][24][25]. Briefly, the expression plasmid obtained from Prof. Tanetoshi Koyama of Tohoku University was used to transform Escherichia coli BL21 (DE3) cells.…”