Substrate Specificities of Wild and Mutated Farnesyl Diphosphate Synthases fromBacillus Stearothermophiluswith Artificial Substrates

Self Cite

We assayed biologically significant isoprenoids for antibacterial and antifungal activities. Isoprenoids varied widely with respect to their growth inhibitory activities towards Gram-positive and Gram-negative bacteria and fungal species. Of particular interest was the finding that geraniol acted as a broad spectrum of antibiotic, and significantly inhibited the growth of Gram-positive Staphylococcus aureus 209P and methicillin resistant S. aureus 834, Gram-negative Escherichia coli IFO-3806, and Shigella sonnei, as well as the fungus Trichopyton sp. Farnesol inhibited S. aureus strains 209P and 834 to a greater extent than did geraniol, but did not detectably inhibit Gram-negative bacteria.

Section: Introductionmentioning

confidence: 99%

Antibacterial and antifungal activities of isoprenoids

Nagaki

Narita

Ichikawa

et al. 2011

Self Cite

“…Relative yields of FPS reactions were then calculated relative to those of FPP derived from the standard enzymatic reaction with IPP and DMAPP. Our materials were similar to those used in previous experiments [16][17][18][19]; our HPLC apparatus was a Hitachi L-6000 (Hitachi, Tokyo, Japan) equipped with a Hitachi LaChrom L-7420 UV-VIS detector and a ChromatoDAQ II (ULVAC, Inc., Chigasaki, Japan). We also used a LichroCART column (Merck-Japan, Tokyo, Japan) with a hexane:2-propanol (40:1) eluent.…”

Section: Discussionmentioning

confidence: 99%

“…[11] The motif (DD**D), which is rich in aspartate (D), is known to be an active site of this enzyme, and tyrosine (Y81) in five upper streams of the first aspartate rich motif (FARM) is believed to be important in product chain length determination [11]. Several studies have investigated the substrate specificities of FPS with allylic and homoallylic substrate homologs [12][13][14][15][16]. We recently reported on the reactivities of several allylic substrate homologs which have a hydrophilic group at the -position, using B. stearothermophilus and porcine liver FPSs [17][18][19].…”

Section: Introductionmentioning

confidence: 99%

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

Musashi

Kanno

Kawakami

et al. 2010

Self Cite

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%).

“…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.…”

Section: Purification Of E-and Z-fpp Synthasesmentioning

confidence: 99%

“…Several previous studies have investigated the substrate specificities of FPP synthase with both allylic and homoallylic substrate homologs [16][17][18][19][20].…”

Section: Introductionmentioning

confidence: 99%

Substrate Specificities of E-and Z-Farnesyl Diphosphate Synthases with Artificial Substrates

Nagaki

Musashi

Kawakami

et al. 2010

Self Cite

Here, to assess substrate specificities of E-and Z-farnesyl diphosphate synthases from Bacillus stearothermophilus and Thermobifida fusca, we examined the reactivities of isopentenyl diphosphate homologs having an alkyl group at the 3-position. E-and Z-FPP synthase reactions of geranyl diphosphate (GPP) with 3-ethylisopentenyl diphosphate produced E-and Z-3-ethylfarnesyl diphosphates (yield: 70.8%, and 11.3%, respectively). The E-FPP synthase reaction of GPP with 3-propylisopentenyl diphosphate produced E-3-propylfarnesyl diphosphate (yield: 40.1%); however, when reacting GPP with 3-propylisopentenyl diphosphate by Z-FPP synthase reaction, only trace product was acquired. Further, neither E-nor Z-FPP synthase reaction of GPP with 3-butylisopentanyl diphosphate produced any product.