Synthesis of 1,2,3,4-tetrahydroxybenzene from D-glucose: Exploiting myo- inositol as a precursor to aromatic chemicals [11]

Full text of DOI:10.1021/ja9840293

J. Am. Chem. Soc. 1999, 121, 3799-3800
3799
Scheme 1a
Synthesis of 1,2,3,4-Tetrahydroxybenzene from
D-Glucose: Exploiting myo-Inositol as a Precursor to
Aromatic Chemicals
Chad A. Hansen, Amy B. Dean, K. M. Draths, and
J. W. Frost*
Department of Chemistry, Michigan State UniVersity
East Lansing, Michigan 48824
ReceiVed NoVember 23, 1998
Polyhydroxy benzenes and quinones possessing the oxygen-
ation pattern of 1,2,3,4-tetrahydroxybenzene 1 often display
biological activity. Aurantiogliocladin 2 and fumigatin 3 are
antibiotics.1 Coenzyme Qn)10 4 is an essential antioxidant in
humans protecting low-density lipoproteins from atherosclerosis-
related oxidative modification.2 Dillapiole 5 is a pyrethrin
synergist and responsible for the sedative effect of Perilla
frutescens leaves.3 A synthetic route (Scheme 1) has now been
elaborated which provides convenient access to 1,2,3,4-tetrahy-
a Key: (a) phosphoenolpyruvate:carbohydrate phosphotransferase; (b)
myo-inositol 1-phosphate synthase; (c) phosphatase activity; (d) dehy-
drogenase activity; (e) 0.5 M H2SO4, H2O, reflux.
Figure 1. Cultivation of E. coli JWF1/pAD1.88A under fed-batch
fermentor conditions: solid bar, inositol; open bar, myo-inositol 1-phos-
phate; (b) cell dry weight.
droxybenzene via myo-inositol intermediacy. The general utility
of this route is demonstrated by a concise synthesis of coenzyme
Qn)3 4. While the shikimate pathway and polyketide biosynthesis
have traditionally provided biocatalytic access to aromatic
chemicals, syntheses of 1,2,3,4-tetrahydroxybenzene 1 and co-
enzyme Qn)3 4 are distinguished by the recruitment of myo-
inositol biosynthesis.
Synthesis of myo-inositol by Escherichia coli JWF1/pAD1.88A
begins with D-glucose uptake and conversion to D-glucose
6-phosphate catalyzed by the E. coli phosphotransferase system4
where phosphoenolpyruvate is the source of the transferred
phosphoryl group (Scheme 1). D-Glucose 6-phosphate then
undergoes cyclization to myo-inositol 1-phosphate catalyzed by
myo-inositol 1-phosphate synthase. This enzyme activity, which
results from expression of the Saccharomyces cereVisiae INO1
gene5 on plasmid pAD1.88A, varied significantly (0.022, 0.043,
0.018, and 0.009 µmol/min/mg at 18, 30, 42, and 54 h,
respectively) over the course of the fermentation.
inositol 1-phosphate to myo-inositol is catalyzed by the enzyme
inositol monophosphatase.6 Phosphoester hydrolysis was fortu-
itously catalyzed in E. coli JWF1/pAD1.88A by unidentified
cytosolic or periplasmic phosphatase activity.
Oxidation of myo-inositol to myo-2-inosose, the next step in
the conversion of D-glucose into 1,2,3,4-tetrahydroxybenzene 1,
is the first catabolic step when myo-inositol is used as a sole source
of carbon for growth and metabolism by microbes such as Bacillus
subtilis.7 myo-Inositol can also be oxidized by Gluconobacter
oxidans without loss of product myo-2-inosose to catabolism.8
Accordingly, incubation of G. oxidans ATCC 621 in medium
containing microbe-synthesized myo-inositol led to the formation
of myo-2-inosose (Scheme 1) in 95% isolated yield.
Inososes have been thought to be stable under acidic conditions
and reactive under basic conditions with reported aromatizations
resulting from successive â-eliminations being dominated by
formation of 1,2,3,5-tetrahydroxybenzene.9 We, however, ob-
served myo-2-inosose to be reactive under acidic conditions with
no apparent formation of 1,2,3,5-tetrahydroxybenzene. Refluxing
G. oxidans-produced myo-2-inosose for 9 h in degassed, aqueous
0.5 M H2SO4 under argon cleanly afforded 1,2,3,4-tetrahydroxy-
benzene in 66% isolated yield.
Conversion of D-glucose into 1,2,3,4-tetrahydroxybenzene 1
is a three-step synthesis. 1,2,3,4-Tetrahydroxybenzene 1 has
historically been obtained from pyrogallol 6 by a longer route
(Scheme 2) involving synthesis and subsequent hydrolysis of
aminopyrogallol 7.10 Due to the tedious nature of this synthesis,10b
two alternate routes (Scheme 2) were used to obtain authentic
E. coli JWF1/pAD1.88A synthesized 21 g/L myo-inositol and
4 g/L myo-inositol 1-phosphate in 11% combined yield (mol/
mol) from D-glucose under fed-batch fermentor conditions (Figure
1). Both myo-inositol and myo-inositol 1-phosphate accumulated
in the culture supernatant. In eucaryotes, hydrolysis of myo-
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10.1021/ja9840293 CCC: $18.00 © 1999 American Chemical Society
Published on Web 04/07/1999

3800 J. Am. Chem. Soc., Vol. 121, No. 15, 1999
Communications to the Editor
Scheme 2a
Scheme 3a
a Key: (a) (CH3)2SO4, NaOH, 69%; (b) (i) n-BuLi, TMEDA, hexanes,
THF, 0 °C; (ii) CH3I, 0 °C, 83%; (c) (i) n-BuLi, TMEDA, hexanes, 0
°C; (ii) CuCN, THF, Et2O, 0 °C; (iii) farnesyl bromide, -78 °C, 57%;
(d) CAN, pyridine-2,6-dicarboxylate, CH3CN/H2O, 0 °C, 46%.
uses p-cresol as a starting material and substantially shorter than
syntheses of coenzyme Qn from pyrogallol, gallic acid, or
vanillin.13b-d
Only one oxygen atom in coenzyme Qn, a shikimate pathway
product, is directly derived from D-glucose. The remaining oxygen
atoms are derived from O2 via enzyme-catalyzed hydroxylations.
Trihydroxybenzenes, pyrogallol and phloroglucinol possess the
maximum number of oxygen atoms attached to a benzene nucleus
by the shikimate pathway or polyketide biosynthesis in lieu of
enzyme-catalyzed hydroxylation. At least a dozen enzymes are
required to disassemble and reassemble the carbon atoms of
D-glucose into the benzene nucleus of coenzyme Qn, pyrogallols,
and phloroglucinols. By comparison, synthesis of 1,2,3,4-tetrahy-
droxybenzene 1 via myo-inositol intermediacy requires only four
enzymes and an acid-catalyzed dehydration for all six carbon and
all four oxygen atoms to be directly derived from the carbon and
oxygen atoms of D-glucose. Synthesis (Scheme 1) of 1,2,3,4-
tetrahydroxybenzene 1 is thus a useful example of enzyme and
atom14 economy in organic synthesis in addition to being a
significant strategic departure from previous biocatalytic syntheses
of aromatic chemicals from D-glucose.
a Key: (a) Cl2C(O), pyridine, xylene, reflux; (b) H2SO4, HNO3; (c)
KOH (aq); (d) Zn, HCl; (e) H2O, reflux; (f) BnBr, K2CO3, acetone, reflux,
83%; (g) K3Fe(CN)6, H2O2, AcOH, 11%; (h) H2, 10% Pd/C, EtOH, 100%;
(i) N-methylformanilide, POCl3, 60 °C, 93%; (j) HCO2H, H2O2, CH2Cl2,
0 °C to room temperature, 95%; (k) H2, 10% Pd/C, EtOH, 80%.
samples of 1,2,3,4-tetrahydroxybenzene 1. Low-yielding, direct
hydroxylation11 of protected pyrogallol 8 or higher-yielding,
indirect oxidation via formyl 10 intermediacy12 yielded respec-
tively quinone 9 and phenol 11.12 Hydrogenation of 9 and 11
afforded products which were identical to 1,2,3,4-tetrahydroxy-
benzene 1 synthesized (Scheme 1) from D-glucose.
Variations in strategies employed for hydroxyl protection
combined with the ease of metalation and alkylation of the
aromatic nucleus makes 1 a versatile intermediate for the synthesis
of a wide spectrum of naturally occurring 1,2,3,4-tetrahydroxy-
benzene derivatives. For example, permethylation (Scheme 3) of
1 leads to tetramethyl 12 which undergoes facile lithiation and
methylation affording 13 in high yield. Formation of an organo-
cuprate from 13, farnesylation, and subsequent reaction with
(NH4)2Ce(NO3)6 affords coenzyme Qn)3 4. This four-step syn-
thesis of coenzyme Qn from tetrahydroxybenzene 1 is equal in
length to the shortest reported13a synthesis of coenzyme Qn which
Acknowledgment. Research was supported by the National Science
Foundation. Professor Susan A. Henry provided the INO1 locus.
Supporting Information Available: Synthesis of myo-inositol, myo-
2-inosose, 1, 8, 9, 10, 11, 12, 13, 14, and 4 (PDF). This material is
available free of charge via the Internet at http://pubs.acs.org.
JA9840293
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