Substrate specificities of several prenyl chain elongating enzymes with respect to 4-methyl-4-pentenyl diphosphate

Article abstract of DOI:10.1271/bbb.68.2070

In order to develop synthetic methods for biologically active homoallylic terpene sulfates, we examined the applicability and substrate specificities of several prenyl chain elongating enzymes with respect to 4-methyl-4-pentenyl diphosphate (homoIPP). The reaction of dimethylallyl diphosphate with homoIPP by use of Bacillus stearothermophilus (all-trans)-farnesyl diphosphate synthase resulted in efficient yields of cis-(yield: 45.9%) and trans-4,8-dimethylnona-3, 7-dien-1-ol (homoGOH, 25.5%), which has a carbon skeleton of 4,8-dimethylnona-3-en-1-sulfate, an antiproliferative compound from a marine organism (Aiello, A. et al., Tetrahedron, 53, 11489-11492 (1997)). The homoIPP was found to be also active as a homoallylic substrate in place of isopentenyl diphosphate for Sulfolobus acidocaldarius geranylgeranyl diphosphate synthase to give diphosphate of cis- and trans-4,8,12-trimethyltrideca-3,7,11-trien-1-ol, for Micrococcus luteus B-P 26 hexaprenyl diphosphate synthase to give cis- and trans-4,8,12,16-tetramethylheptadeca-3,7,11,15-tetraen-1-ol (homoGGOH), and for Micrococcus luteus B-P 26 undecaprenyl diphosphate synthase to give cis-homoGGOH exclusively.

Full text of DOI:10.1271/bbb.68.2070

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Substrate Specificities of Several Prenyl Chain
Elongating Enzymes with Respect to 4-Methyl-4-
pentenyl Diphosphate
b
a
a
a
a
Masahiko NAGAKI , Yohei MIKI , Minori NAKADA , Jun KAWAKAMI , Haruo KITAHARA , Yuji
c
c
d
e
MAKI , Yoshinori GOTOH , Tokuzo NISHINO & Tanetoshi KOYAMA
a
Department of Materials Science and Technology, Faculty of Science and Technology,
Hirosaki University
b
Department of Natural Science, Faculty of Education, Hirosaki University
c
Department of Biological and Material Chemistry, Faculty of Science, Yamagata
University
d
Department of Biochemistry and Engineering, Faculty of Engineering, Tohoku University
e
Institute of Multidisciplinary Research for Advanced Materials, Tohoku University
Published online: 22 May 2014.
To cite this article: Masahiko NAGAKI, Yohei MIKI, Minori NAKADA, Jun KAWAKAMI, Haruo KITAHARA, Yuji MAKI, Yoshinori
GOTOH, Tokuzo NISHINO & Tanetoshi KOYAMA (2004) Substrate Specificities of Several Prenyl Chain Elongating Enzymes
with Respect to 4-Methyl-4-pentenyl Diphosphate, Bioscience, Biotechnology, and Biochemistry, 68:10, 2070-2075, DOI:
10.1271/bbb.68.2070
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Biosci. Biotechnol. Biochem., 68 (10), 2070–2075, 2004
Substrate Specificities of Several Prenyl Chain Elongating Enzymes
with Respect to 4-Methyl-4-pentenyl Diphosphate
1
;y
1
1
1
2
Masahiko NAGAKI, Yohei MIKI, Minori NAKADA, Jun KAWAKAMI, Haruo KITAHARA,
3
3
4
5
Yuji MAKI, Yoshinori GOTOH, Tokuzo NISHINO, and Tanetoshi KOYAMA
1
Department of Materials Science and Technology, Faculty of Science and Technology,
Hirosaki University, 3 Bunkyo-cho, Hirosaki, Aomori 036-8561, Japan
2
Department of Natural Science, Faculty of Education, Hirosaki University,
1
Bunkyo-cho, Hirosaki, Aomori 036-8560, Japan
3
Department of Biological and Material Chemistry, Faculty of Science, Yamagata University,
Kojirakawa-machi, Yamagata 990-8560, Japan
4
Department of Biochemistry and Engineering, Faculty of Engineering, Tohoku University,
Aoba-ku, Sendai, Miyagi 980-8579, Japan
5
Institute of Multidisciplinary Research for Advanced Materials, Tohoku University,
Aoba-ku, Sendai, Miyagi 980-8577, Japan
Received May 6, 2004; Accepted July 5, 2004
In order to develop synthetic methods for biologically
active homoallylic terpene sulfates, we examined the
applicability and substrate specificities of several prenyl
chain elongating enzymes with respect to 4-methyl-4-
pentenyl diphosphate (homoIPP).
tetraen-1-ol (homoGGOH), and for Micrococcus luteus
B-P 26 undecaprenyl diphosphate synthase to give cis-
homoGGOH exclusively.
Key words: prenyl chain elongating enzyme; farnesyl
diphosphate synthase; geranylgeranyl di-
phosphate synthase; undecaprenyl diphos-
phate synthase; substrate specificity
The reaction of dimethylallyl diphosphate with ho-
moIPP by use of Bacillus stearothermophilus (all-trans)-
farnesyl diphosphate synthase resulted in efficient yields
of cis-(yield: 45.9%) and trans-4,8-dimethylnona-3,7-
dien-1-ol (homoGOH, 25.5%), which has a carbon
skeleton of 4,8-dimethylnona-3-en-1-sulfate, an antipro-
liferative compound from a marine organism (Aiello,
A. et al., Tetrahedron, 53, 11489–11492 (1997)). The
homoIPP was found to be also active as a homoallylic
substrate in place of isopentenyl diphosphate for
Sulfolobus acidocaldarius geranylgeranyl diphosphate
synthase to give diphosphate of cis- and trans-4,8,12-
trimethyltrideca-3,7,11-trien-1-ol, for Micrococcus lu-
teus B-P 26 hexaprenyl diphosphate synthase to give
cis- and trans-4,8,12,16-tetramethylheptadeca-3,7,11,15-
The enzymatic prenyl chain elongating reaction
proceeds with the condensation of an allylic primer
substrate with isopentenyl diphosphate (IPP, 1a) stereo-
specifically. The condensation terminates precisely
when the elongation of the prenyl chain reaches a
certain length according to the specificities of individual
synthases, as shown in Scheme 1. These prenyl chain
elongating enzymes can be classified into four classes
according to mode of subunit composition as well as
chain length and the stereochemistry of the final
1
–3)
products.
Scheme 1. Prenyl Chain Elongating Enzymatic Reactions.
y
To whom correspondence should be addressed. Fax: +81-172-39-3947; E-mail: nagaki@cc.hirosaki-u.ac.jp
Abbreviations: FPP, farnesyl diphosphate; IPP, isopentenyl diphosphate; UPP, undecaprenyl diphosphate; DMAPP, dimethylallyl diphosphate;
GPP, geranyl diphosphate

Subtrate Specificities of Prenyl Chain Elongating Enzymes
2071
Scheme 2. Prenyl Chain Elongating Enzymatic Reactions between Allylic Substrate and HomolPP.
Porcine lever FPP synthase has been shown to produce 4a or 4b from the reaction with DMAPP or GPP, respectively.7,8)
80:1 (v=v) (B). Identification of the reaction products
was carried out with a GC-MS, JMS-AM II 50 type
GCG Mass spectrometer connected with a HP 5890
series II Gas chromatograph equipped with an Ultra-
alloy-1 (S) column. The column temperature was
ꢀ
programmed from 90 to 280 C with a linear gradient
of temperature at a rate of 15 C/min and then held at
Scheme 3. Sodium cis-4,8-Dimethylnona-3-en-1-sulfate, an Antipro-
liferative Terpene from a Marine Organism.9)
ꢀ
ꢀ
280 C for 3 min. The relative yields of the prenyltrans-
ferase reactions, except for those of FPP synthase
reactions, were calculated on the basis of the amount of
recovered farnesol derived from the residual substrate
FPP in the reaction mixture. The relative yields of FPP
synthase reactions were those of the homologs of GPP
and FPP derived from reactions with a set of substrates
compared to the yield of FPP derived from the reaction
with IPP and DMAPP or with IPP and GPP, respec-
tively.
From the standpoint of application of the enzyme in
organic syntheses, prenyl chain elongating enzymes
have been shown to be useful in stereospecific syntheses
of some kinds of biologically active compounds. Porcine
liver farnesyl diphosphate (FPP) synthase reaction has
4
)
been applied in the chiral syntheses of faranal and 4-
5
)
methyljuvenile hormone. Undecaprenyl diphosphate
(UPP) synthase from Escherichia coli has been applied
to the one-pot synthesis of glycinoprenols.
6
)
7
,8)
Ogura et al.
reported that 4-methyl-4-pentenyl
Chemicals.
Synthesis of IPP-homolog. 4-Methyl-4-pentenyl
diphosphate7,10) (homoIPP) was synthesized by di-
diphosphate (homoIPP, 1b) was accepted as a substrate
in place of the homoallylic substrate, IPP, for a porcine
liver FPP synthase, and that the product derived from the
reaction of homoIPP with GPP (2b) or with DMAPP
1
1)
phosphorylation of 4-methyl-4-pentene-1-ol (yield:
47.0%), which was prepared by reduction of 4-methyl-
1
2)
(
2a) was exclusively cis-homofarnesyl-(4b) or cis-
4-pentenoic acid with LiAlH4.
homogeranyl diphosphate (4a), respectively, as shown
in Scheme 2.
Chemical synthesis of an authentic sample of 4,8,12,
16-tetramethylheptadeca-3,7,11,15-tetraen-1-ol (homo-
GGOH). Cyanation with sodium cyanide of geranylger-
anyl bromide, which was derived from bromination of
geranylgeraniol by the use of phosphorus bromide, gave
4,8,12,16-tetramethylheptadeca-3,7,11,15-tetraenonitrile
(homogeranylgeranyl cyanide) (yield: 74.0%). 4,8,12,
16-tetramethylheptadeca-3,7,11,15-tetraenoic acid (ho-
mogeranylgeranoic acid), which was derived from
hydrolysis of homogeranylgeranyl cyanide, was reduced
by the use of lithium aluminum hydride to give a 3-cis
and 3-trans mixture of 4,8,12,16-tetramethylheptadeca-
Recently, antiproliferative terpene derivatives, which
have homoallylic carbon skeletons such as cis-4,8-
dimethylnona-3-en sodium sulfate (5), have been iso-
9
)
lated from marine organisms such as the sea squirt, as
shown in Scheme 3.
In this study, in order to develop synthetic methods
for this kind of biologically active homoterpene com-
pounds, we examined the applicability and substrate
specificities of several prenyl chain elongating enzymes
with respect to an artificial IPP homolog, homoIPP.
3
,7,11,15-tetraen-1-ol (homoGGOH, yield: 16.1%). The
1
Materials and Methods
H NMR (400 MHz, CDCl3, TMS) of the cis- and trans
mixture of homoGGOH was constituted as follows: ꢀ:
1.64 (16H, s.), 2.06 (12H, dt.), 2.29 (2H, dt.), 3.62
(2H,t), 5.10 (4H, s). The C NMR (100 MHz, CDCl3,
TMS) of the cis- and trans mixture of homoGGOH was
constituted as follows: ꢀ: 15.9, 16.0, 17.7, 23.4, 25.7,
26.5, 26.6, 26.7, 27.5, 27.6, 31.9, 32.4, 39.7, 39.9, 63.8,
119.8, 123.6, 124.2, 124.4, 124.9, 129.0, 131.3, 135.0,
Analysis. Prenyl alcohols derived from the phospha-
tase treatment of products from prenyltransferase reac-
tions were subjected to HPLC, a Hitachi type L-6000
equipped with Hitachi L-7420 (LaChrom) type UV–VIS
detector with a Chromato Monitor (Nippon Filcon) and
with a LichroCART (Merck) column with the eluents of
solvent mixtures of hexane:2-propanol = 40:1 (A), or
1
3
ꢁ
1
138.9. And the IR ꢁmax (KBr) cm of homoGGOH was

2072
M. NAGAKI et al.
constituted as follows: 3352 (OH), 1048 (C–O). It has
been shown for these homoprenyl alcohols that under
the GC conditions of these homologs, the cis-isomers
have a shorter retention time than that of the corre-
sponding trans-isomer. The retention times of the
cis- and trans-homoGGOH on GC were 12.2 min and
FPP synthase solution (25 ꢄg, 100 ꢄl) was used. After
ꢀ
incubation at 55 C for 6 h, the reaction mixture was
treated with alkaline phosphatase for 5 h, and the
reaction products were extracted with pentane and
13)
analyzed by HPLC and GC-MS.
Mesophilic enzymatic reactions. The incubation mix-
ture for M. luteus B-P 26 HexPP synthase reaction
contained, in a total volume of 1.0 ml, 50 mM of Tris–
1
3)
1
2.3 min respectively, and the area ratio of these peaks
was 3:7. The MS spectral data of cis-homoGGOH were
þ
þ
as follows: m=z: 304 (M ) (rel. int. 0.4%), 286 (M ꢁ
HCl buffer (pH 8.5), 1.0 mM of MgCl , 20 mM of ꢃ-
2
þ
þ
1
6
8) (1.7), 217 (M ꢁ 18 ꢁ 69) (2.4), 149 (M ꢁ 18 ꢁ
mercaptoethanol, 50 mM of NH Cl, 1.0 mM of FPP,
4
þ
9 ꢁ 68) (8.8), 81 (M ꢁ 18 ꢁ 69 ꢁ 68) (54.1), 69 (base
1.0 mM of IPP or homoIPP, and the recombinant HexPP
synthase (76 and 90 ꢄg of components A and -B,
peak). And the MS spectral data of trans-homoGGOH
þ
20)
were as follows: m=z: 304 (M ) (rel. int. 0.7%), 286
þ
respectively). HexPP synthase consists of two dis-
sociable heterodimeric components A and -B, each of
which lacks catalytic activity but is activated when
þ
þ
(
1
M ꢁ 18) (1.9), 217 (M ꢁ 18 ꢁ 69) (2.5), 149 (M ꢁ
þ
8 ꢁ 69 ꢁ 68) (9.2), 81 (M ꢁ 18 ꢁ 69 ꢁ 68) (54.2), 69
1
9)
(
base peak).
Other homologs such as homogeraniol were synthe-
combined with the other. The M. luteus B-P 26 UPP
synthase reaction was executed under similar conditions,
2
1)
sized similarly according to the synthetic method
reported.8
as described previously. The mixture was incubated at
ꢀ
)
37 C for 6 h. After incubation, the reaction mixture was
treated with 1-butanol and the products were hydrolyzed
Potato acid phosphatase was purchased from Boeh-
ringer Mannheim (Roche, grade II, catalog no. 108 227).
ꢀ
with acid phosphatase at 37 C for 12 h. The hydro-
lysates were extracted with pentane and analyzed by
HPLC and GC-MS.
Purification of prenyl chain elongating enzymes.
Purification of S. acidocaldarius GGPP synthase.
E. coli DH5ꢂ cells were transformed with pUC119
plasmid that contained an insert of the S. acidocaldarius
Results and Discussion
1
4)
GGPP synthase gene.
inoculate LB medium containing ampicillin and incu-
The colonies were used to
In order to develop synthetic methods for biologically
active terpenes such as antiproliferative compounds, we
examined the substrate specificities with respect to
homoIPP of several prenyl chain elongating enzymes.
ꢀ
bated at 37 C overnight. The cells were harvested and
disrupted by sonication. The homogenate was heated at
55 C for 60 min and then centrifuged at 10;000 ꢂ g.
The supernatant fraction of E. coli DH5ꢂ/pGGPS3 was
ꢀ
Reaction of DMAPP or GPP with homoIPP by use of
FPP synthase
precipitated with a 30–60% saturation of (NH ) SO .
4
2
4
The precipitated protein fraction was dialyzed and
purified by two chromatographies (Butyl-Toyopearl
Reaction of DMAPP (2a) with homoIPP (1b) by use of
FPP synthase. The hydrolysates derived from the
product of the B. stearothermophilus FPP synthase
reaction of 1b with DMAPP as substrate gave two
peaks on HPLC (eluent B) at 29.3 and 31.1 min, which
were then subjected to GC-MS. The MS spectrum of the
former alcohol showed a molecular ion at m=z 168
(rel. int. 2.3%), corresponding to C H O, together with
and MonoQ).1
4–16)
The purities of the GGPP synthase
fractions were analyzed by sodium dodecyl sulfate-
polyacrylamide gel electrophoresis after staining with
Coomassie Brilliant Blue.
Purification of other enzymes. B. stearothermophilus
or porcine liver FPP synthase was purified according to
1
1
20
þ
1
3,17,18)
the method reported previously.
synthases from M. luteus B-P 26 were purified in the
HexPP- and UPP
main fragment ions at m=z 150 (M ꢁ 18) (4.6), 81
þ
(M ꢁ 18 ꢁ 69) (40.9), and 69 (base peak), indicating
similar manner, as reported previously.19,20)
that the alcohol has a 4,8-dimethylnona-3,7-dien-1-ol
(homogeraniol) structure. On the other hand, the
Conditions of the enzyme-catalyzed reaction.
spectrum of the latter alcohol showed the same molecu-
lar ion as well as similar fragment ions with some
differences in relative intensity, which also indicates a
homogeraniol structure.
Thermostable enzyme-catalyzed reactions. The incu-
bation mixture for the S. acidocaldarius GGPP synthase
reaction contained, in a total volume of 1.0 ml, 100 mM
of phosphate buffer (pH 5.8), 2.0 mM of MgCl , 10 mM
In order to determine the geometry of the double bond
at the 3-position, authentic cis- and trans-homogeraniol
were synthesized from geranyl bromide via correspond-
ing 4,8-dimethylnona-3,7-dienoic acid by lithium alu-
minum hydride reduction.
2
of ꢃ-mercaptoethanol, 10 mM of NH Cl, 1.0 mM of an
4
allylic substrate (GPP or FPP), 1.0 mM of IPP or
homoIPP, and the recombinant GGPP synthase solution
(
200 ꢄl). The B. stearothermophilus FPP synthase re-
action was executed under conditions similar to those
described for the GGPP synthase, except that 200 mM of
Tris–HCl buffer (pH 8.5), 1.0 mM of IPP or homoIPP,
The authentic cis- and trans-homogeraniol eluted on
HPLC (eluent A) had retention times of 16.2 and
17.4 min, respectively. These results indicate that the
former, derived from the enzymatic reaction product, is
1
.0 mM of an allylic substrate (DMAPP or GPP), and

Subtrate Specificities of Prenyl Chain Elongating Enzymes
2073
assignable to cis-homogeraniol (4a-OH), and the latter
to the trans-isomer (3a-OH). The relative yields of 4a
and 3a were 45.9% and 25.5% of that of GPP derived
from the reaction with DMAPP and IPP.
Our re-experiment on the action of porcine liver FPP
synthase-catalyzed reaction with IPP homolog (1b)
confirmed exclusive formation of the cis-isomer, al-
though enzyme activity was as low as 1% compared
with that of the thermostable enzyme.
Reaction of FPP (2c) with homoIPP (1b) by use of
S. acidocaldarius GGPP synthase. The alcohols derived
from phosphatase treatment of the products of the
S. acidocaldarius GGPP synthase reaction of 1b with
FPP gave two peaks on GC at 12.1 (relative yield: 6.3%)
and 12.2 min (relative yield: 11.2%). These two products
were found to be identical with those of authentic
cis- and trans-homogeranylgeraniol respectively. Hence
the former was identified as cis-(4c-OH) and the latter
as trans-homogeranylgeraniol (3c-OH).
In 1997, Aiello et al. reported the isolation of an
antiproliferative terpene sulfate from a Mediterranean
9)
ascidian. This biologically active terpene has the same
carbon skeleton to that of cis-homogeraniol. From the
viewpoint of the possibility of synthetic application of
FPP synthase to obtain the carbon skeleton of the
antiproliferative terpene, the B. stearothermophilus FPP
synthase appears to be more useful because of the large
yield compared to that of the liver enzyme.
Reaction of FPP with homoIPP by use of HexPP
synthase or UPP synthase
Reaction of FPP (2c) with homoIPP (1b) by use of
M. luteus B-P 26 HexPP synthase. The alcohols derived
from the products of the M. luteus B-P 26 HexPP
synthase reaction of 1b with FPP gave two peaks on GC
at 12.2 (relative yield: 3.5%) and 12.3 min (relative
yield: 5.2%). Both of the MS spectra of these products
exactly coincided with the authentic cis-(4c-OH) and
trans-homogeranylgeraniol (3c-OH) respectively.
Reaction of FPP (2c) with homoIPP (1b) by use of
M. luteus B-P 26 UPP synthase. The hydrolysate
derived from acid phosphatase treatment of the product
of the M. luteus B-P 26 UPP synthase reaction of 1b
with FPP gave a peak on GC at 12.2 min (relative yield:
2.0%). The GC-MS spectrum of the product gave a
similar MS spectrum to that of cis-homogeranylgeraniol
(4c-OH), indicating that the enzymatic product was
cis-geranylgeranyl diphosphate (4c).
Reaction of GPP with homoIPP by use of FPP
synthase. The alcohols derived from the reaction of the
thermostable FPP synthase with homoIPP (1b) and GPP
(
(
2b) as substrates eluted on HPLC as two peaks at 35.5
relative yield: 2.9%) and 36.9 min (relative yield:
2
.6%), which were then purified and subjected to GC-
MS. In the MS spectrum of the former product, the
molecular ion was observed at m=z 236 (rel. int.: 5.0%),
corresponding to C H O, with other fragment ions
16
28
þ
þ
at m=z 218 (M ꢁ 18) (5.4), 149 (M ꢁ 18 ꢁ 69) (8.9),
þ
8
1 (M ꢁ 18 ꢁ 69 ꢁ 68) (62.4), and 69 (base peak),
indicating that the alcohol has a 4,8,12-trimethyltrideca-
,7,11-trien-1-ol (homofarnesol) structure. The latter
3
product also showed a molecular ion at m=z 236 (rel.
int.: 2.7%), and other fragment ions similar to those of
the former product, at m=z 218, 149, 81, and 69, for
example, suggesting that the product has a homofarnesol
structure.
As mentioned above, cis-homogeraniol eluted on the
HPLC with a shorter retention time than that of the
corresponding trans-isomer. The former product was
assigned to cis-homofarnesol (4b-OH) and the latter to
the trans-isomer (3b-OH).
Conclusion
Scheme 2 illustrates the prenyl chain elongating
reactions with respect to homoIPP as the non-allylic
substrate catalyzed by the prenyltransferases examined
in this study. The relative yields of the homoallylic
alcohols obtained by phosphatase treatment of each
prenyltransferase reaction product are shown in Table 1.
By obtaining 3-cis-4,8-dimethylnona-3,7-dien-1-ol
(4a-OH) derived from a large-scale enzymatic reaction
by thermostable FPP synthase, it could be possible to
synthesize the antiproliferative homoterpene sulfate by
When porcine liver FPP synthase was used, the cis-
isomer (4b-OH, relative yield: 7%) was obtained
7
)
22)
exclusively, as reported previously.
Sharpless epoxidation of the corresponding THP ether
3)
2
obtained by the use of PPTS for the protection of the
double bond at C-3 and by hydrogenation at C-7, and
Reactions of GPP or FPP with homoIPP by use of
GGPP synthase
2
4,25)
then reduction of epoxide to olefine via iodohydrin,
2
6)
Reaction of GPP (2b) with homoIPP (1b) by use of
S. acidocaldarius GGPP synthase. The hydrolysate
derived from acid phosphatase treatment of the product
of the S. acidocaldarius GGPP synthase reaction of 1b
with GPP gave two peaks at 29.4 (relative yield: 17.6%)
and 30.0 min (relative yield: 28.2%) on HPLC (eluent
B). These were subjected to GC-MS analysis. The two
alcohols showed similar MS spectra to those of the
homofarnesols, 3b-OH and 4b-OH, indicating that the
former is the cis-homofarnesol (4b-OH) and the latter
the trans-isomer (3b-OH).
followed by sulfuric esterification.
Acknowledgment
This work was supported by Grant-in-Aid for Scien-
tific Research 12680586 (to M.N.) from the Ministry
of Education, Culture, Sports, Science and Technology
of Japan. Part of this work was supported by the
Kuribayashi Education Foundation and the Asahi Glass
Foundation.

2074
M. NAGAKI et al.
Table 1. Relative Yields of the Homoallylic Alcohols Derived from Prenyltransferase Reactions with HomoIPP as Non-allylic Substrate
ꢃ
Product homoallylic alcohols derived from phosphatase treatment of each enzymatic product (%)
Prenyltransferase
FPP synthase
Allylic substrate
3
aOH
3bOH
3cOH
4aOH
4bOH
4cOH
DMAPP
GPP
n.d.
—
—
n.d.
—
—
—
0.1
—
—
7.4
—
—
—
—
—
—
6.3
(porcine liver)
FPP synthase
B. stearothermophilus)
DMAPP
GPP
GPP
25.5
—
—
—
45.9
—
(
2.6
1.7
—
2.9
1.1
—
GGPP synthase
S. acidocaldarius)
—
—
—
11.2
—
—
(
FPP
HexPP synthase
M. luteus B-P 26)
FPP
FPP
—
—
—
—
5.2
—
—
—
—
3.5
2.0
(
UPP synthase
(
n.d.
M. luteus B-P 26)
ꢃ
The relative yields of product alcohols derived from reactions by FPP synthase were relative to the yields of FOH derived from the corresponding enzymatic
reactions with IPP and DMAPP (or GPP). The relative yields of product alcohols derived from reactions by GGPP synthase were relative to the yields of GGOH
derived from the corresponding enzymatic reactions with IPP and FPP. The relative yields of product alcohols derived from reactions by HexPP synthase or UPP
synthase were calculated on the basis of the amount of recovered farnesol derived from the residual substrate FPP in the reaction mixture.
n.d., not detected.
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