A convenient gram-scale synthesis of uridine diphospho(13C6)glucose

Article abstract of DOI:10.1016/j.carres.2006.02.033

A simple gram-scale synthesis of uridine diphospho(¹³C₆)glucose is presented from d-(¹³C₆)glucose. The critical step uses a 1H-tetrazole-catalyzed coupling of 2,3,4,6-tetra-O-acetyl-α-d-glucopyranosyl-1-phosphat

Full text of DOI:10.1016/j.carres.2006.02.033

Carbohydrate Research 341 (2006) 1743–1747
Note
A convenient gram-scale synthesis of uridine diphospho(13C6)glucose
Zoran Dinev,a,b Ahmad Z. Wardak,c Robert T. C. Brownleed and Spencer J. Williamsa,b,
*
aSchool of Chemistry, University of Melbourne, Parkville, Victoria 3010, Australia
bBio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Victoria 3010, Australia
cDepartment of Biochemistry, La Trobe University, Melbourne, Victoria 3086, Australia
dDepartment of Chemistry, La Trobe University, Melbourne, Victoria 3086, Australia
Received 3 February 2006; received in revised form 20 February 2006; accepted 24 February 2006
Available online 5 April 2006
Abstract—A simple gram-scale synthesis of uridine diphospho(13C6)glucose is presented from D-(13C6)glucose. The critical step uses
a 1H-tetrazole-catalyzed coupling of 2,3,4,6-tetra-O-acetyl-a-D-glucopyranosyl-1-phosphate and UMP-morpholidate. The uridine
diphospho(13C6)glucose was used in the structural identification of (1!3)-b-D-glucan from Lolium multiflorum.
ꢀ 2006 Elsevier Ltd. All rights reserved.
Keywords: Sugar nucleotides; UDP-glucose; Morpholidate coupling; Isotopic labeling; Glucan
Sugar nucleotides are the primary building blocks from
which most complex carbohydrates are built.1 Despite
the central importance of sugar nucleotides, there is still
a need for simple practical routes for their preparation.
Here we report a simple high-yielding route for the
preparation of uridine diphospho(13C6)glucose 6 from
D-(13C6)glucose on a gram scale, for use in 13C NMR
and mass spectrometric structural studies of glycans.2–5
Unlike many of the methods used for the synthesis of su-
gar nucleotides that are described in the literature, this
approach relies on the coupling of a protected glucose-
1-phosphate derivative, thereby allowing purification
of intermediates on silica gel.
Enzymatic methods for the synthesis of sugar nucleo-
tides hold great promise for simple preparative
approaches to sugar nucleotides.6 However, despite the
apparent simplicity of these approaches, there remain
significant problems including the difficulty of acquiring
many of the necessary enzymes, the problem of scale-up,
and the difficulty of isolation of the products. For exam-
ple, Ma and Sto¨ckigt have reported a 450 mg enzymatic
synthesis of UDP-(4-13C)glucose from D-(4-13C)glucose;
however, this method required repeated column and
preparative thin layer chromatography and thus is not
amenable to scale-up.7
Synthetic methods, while superficially cumbersome,
are in many cases more reliable for the preparation of
sugar nucleotides. In general, there are two main
approaches: condensation of a sugar and a nucleotide
diphosphate,8–11 or condensation of a sugar phosphate
with an activated nucleotide monophosphate.12,13 The
latter is more widely used as issues of stereochemical con-
trol in the former can lead to anomeric mixtures.9–11
Most commonly the condensation of an unprotected
sugar phosphate with a nucleoside phosphomorpholi-
date under 1H-tetrazole catalysis is used.14 Indeed, Fair-
weather and co-workers have reported the synthesis of
UDP-(13C6)glucose using this standard method.5 How-
ever, this approach requires troublesome purification of
the highly polar unprotected sugar nucleoside diphos-
phate from a complex reaction mixture. In a patent dis-
closure, Oehrlein and Baisch have reported the carbonyl
diimidazole-promoted coupling of acetylated sugar
phosphates with nucleoside monophosphates, followed
by their deprotection using orange peel pectinesterase.15
This method demonstrated the advantages of coupling
protected sugar phosphates, but failed to exploit the
advantageous physical properties of the acetylated prod-
uct in the isolation step. Kosma and co-workers have
*
Corresponding author. Tel.: +61 3 8344 2422; fax: +61 3 9347
8124; e-mail: sjwill@unimelb.edu.au
0008-6215/$ - see front matter ꢀ 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2006.02.033

1744
Z. Dinev et al. / Carbohydrate Research 341 (2006) 1743–1747
published a simple but significant extension to these
methods that involves coupling of acetylated sugar phos-
phates with a nucleoside phosphomorpholidate,16–18
where the acetylated sugar nucleotide was purified by
flash chromatography prior to deacetylation. While very
promising, this method has not been applied on a large
scale and the utility of 1H-tetrazole as a catalyst for pro-
moting the coupling of acetylated phosphosugars and
morpholidates has not been widely investigated.19 Here,
we report on a simple practical route to UDP-(13C6)glu-
cose using 1H-tetrazole-catalyzed coupling of an acetyl-
ated sugar phosphate with a morpholidate as the key
step, that proceeds in an excellent overall yield from
D-(13C6)glucose.
and the reaction cleanly went to completion overnight.
The coupled product was purified by flash chromatogra-
phy to afford the tetraacetate 5 in excellent yield (84%
over two steps) (Scheme 1). The overnight reaction time
and excellent yield seen here stand in contrast to the 1H-
tetrazole-catalyzed morpholidate couplings reported by
the Schmidt group, which required 3 days to achieve
low to moderate yields (13–30%) of UDP-exo-glycal
derivatives.19 Finally, deprotection of 5 was carried
out by treatment of tetraacetate 6 with triethylammo-
nium bicarbonate (TEAB) in methanol/water at
ꢀ20 ꢁC. The product was purified by Biogel chromato-
graphy to afford uridine diphospho(13C6)glucose 6 as
the ammonium salt (Scheme 2). Sugar nucleotide 6
was contaminated with trace amounts of uridine 50-
monophosphate (UMP) and (13C6)glucose 1,2-cyclic
phosphate (ꢁ5%), which arise spontaneously during
the prolonged deprotection conditions. The presence
of these impurities did not affect combustion analysis
of 6, nor its use in biological studies (see below). UMP
has been found to act as a potent inhibitor of many
glycosyltransferases and if its removal is necessary it is
noted that UMP can be hydrolyzed by treatment with
alkaline phosphatase.
D-(13C6)Glucose was acetylated using catalytic sulfu-
ric acid in acetic anhydride to afford the pentaacetate
1. The pentaacetate 1 was treated with hydrazine acetate
to afford hemiacetal 2. The hemiacetal was condensed
with diphenylphosphoryl chloride according to the pro-
cedure of Sabesan and Neira to afford the anomeric
phosphate 3.20 The diphenyl groups of 3 were cleaved
by hydrogenolysis using platinum oxide under hydrogen
and the resulting phosphoric acid 4 was coupled with
uridine 50-monophosphoromorpholidate in pyridine,
according to the procedure of Kosma.18 In our hands,
this procedure was sluggish and even after several days
significant starting material remained. Upon addition
of 1H-tetrazole significant rate acceleration was noted
The utility of UDP-(13C6)glucose 6 for the study of
polysaccharide synthesis and for structural analysis of
the resultant product was illustrated by its use as sub-
strate in reaction with a (1!3)-b-D-glucan synthase
OAc
O
OAc
O
OH
O
a
b
AcO
AcO
AcO
AcO
HO
HO
OAc
OH
OH
AcO
AcO
HO
13C6)glucose
1
2
(
OAc
O
O
N
OAc
O
AcO
AcO
e
c
NH
O
O
AcO
AcO
AcO
O P O P
O
O
AcO
OPO(OR)2
O
OH OH
3: R = Ph
4: R = H
5
d
OH OH
=
13C
Scheme 1. Reagents and conditions: (a) Ac2O, H2SO4, rt, 96%; (b) NH2NH2ÆAcOH, DMF, 91%; (c) (PhO)2POCl, DMAP, 83%; (d) H2, PtO2,
EtOAc/CH3CH2OH (1:1); (e) uridine 50-monophosphoromorpholidate 4-morpholine-N,N0-dicyclohexylcarboxamide salt, 1H-tetrazole, pyridine,
84% over two steps.
OAc
O
OH
O
O
N
O
N
AcO
AcO
HO
HO
NH
NH
O
O
O
O
a
AcO
HO
O P O P
O
O P O P
O
O
O
O
OH OH
O
O
O
+
2. NH4
=
13C
OH OH
OH OH
6: UDP(13C6)Glc
5
Scheme 2. Reagents and conditions: (a) TEAB, CH3OH, H2O, ꢀ20 ꢁC, 94%.

Z. Dinev et al. / Carbohydrate Research 341 (2006) 1743–1747
1745
C1
C3
C5 C2
C4
C6
ppm
100
90
80
70
60
Figure 1. 100.59 MHz 13C NMR spectrum of ꢁ25 lg (1!3)-b-D-glucan prepared from Lolium multiflorum by in vitro labeling with UDP-
(
13C6)glucose (6).
preparation from suspension-cultured Italian ryegrass
(Lolium multiflorum) cells.21 The product (ꢁ25 lg) was
recovered using the procedure of Him et al.22 The 13C
NMR spectrum of the product (Fig. 1) is identical to
that from a similar 13C-labeled (1!3)-b-D-glucan prep-
aration from blackberry (Rubus fruticulosus),5 and
agrees with the characteristic shifts seen for a natural
abundance 13C NMR spectrum of a (1!3)-b-D-
glucan.23
To conclude, a simple chemical synthesis of UDP-
(13C6)glucose 6 is described, with the critical step being
a 1H-tetrazole-catalyzed coupling of the acetylated glu-
cose-1-phosphate 1 and uridine 50-monophosphoromor-
pholidate. Starting with 2 g of D-(13C6)glucose over 1 g
of labeled UDP-(13C6)glucose 6 was synthesized in an
overall yield of 50%. This route is scaleable and should
be readily applied to even larger scale syntheses, or to
the synthesis of other isotopically labeled derivatives.
performed by Sally Duck at the Chemistry Department,
Monash University. D-(13C6)Glucose and hydrazine acet-
ate were obtained from Sigma–Aldrich. Elemental anal-
ysis was performed by C.M.A.S. (Belmont, Victoria).
1.2. Diphenyl 2,3,4,6-tetra-O-acetyl-a-D-(13C6)gluco-
pyranosyl phosphate (3)
1.2.1.
(1).
1,2,3,4,6-Penta-O-acetyl-D-(13C6)glucopyranose
suspension of D-(13C6)glucose (2.00 g,
A
10.7 mmol) in Ac2O (11 mL) was treated with H2SO4
(100 lL) at 0 ꢁC and the mixture was stirred at rt for
2 h. Water (40 mL) was then added followed by EtOAc
(50 mL). The organic phase was separated and washed
with water (3 · 50 mL) and satd aq NaHCO3
(3 · 50 mL), dried (MgSO4), and concentrated in vacuo
to give 1,2,3,4,6-penta-O-acetyl-D-(13C6)glucopyranose
(1) as a clear oil (4.07 g, 96%, b:a ratio = 4:5). 1H
NMR (400 MHz, CDCl3): d 2.01 (s, 3H, CH3b), 2.02
(s, 3H, CH3a), 2.03 (s, 3H, CH3b), 2.04 (s, 3H,
CH3a), 2.05 (s, 3H, CH3a), 2.08 (s, 3H, CH3b), 2.09
(s, 3H, CH3a), 2.12 (s, 3H, CH3b), 2.18 (s, 3H, CH3a),
2.22 (s, 3H, CH3b), 3.62–5.74 (m, 6H, H-2a,b, 3a,b,
4a,b, 5a,b, 6aa,b, 6ba,b), 5.71 (dd, JC,H = 165 Hz,
J1,2 = 9.2 Hz, 1H, H-1b), 6.33 (dm, JC,H = 175 Hz, 1H,
H-1a).
1. Experimental
1.1. General methods
Petroleum spirits refer to a mixed fraction boiling at
40–60 ꢁC. Thin layer chromatography (TLC) was per-
formed with Merck Silica Gel 60 F254, using mixtures
of petroleum spirits–EtOAc unless otherwise stated.
Detection was effected by either charring in a mixture
of 5% sulfuric acid–CH3OH and/or by visualization in
UV light. NMR spectra were obtained on Varian Inova
400 MHz instruments (Melbourne, Australia). Flash
chromatography was performed according to the meth-
od of Still et al. with Merck Silica Gel 60, using adjusted
mixtures of EtOAc–petroleum spirits unless otherwise
stated.24 CH2Cl2 and pyridine were dried over CaH2.
Solvents were evaporated under reduced pressure using
a rotary evaporator. High resolution mass spectra were
1.2.2.
2,3,4,6-Tetra-O-acetyl-D-(13C6)glucopyranose
(2). A solution of 1,2,3,4,6-penta-O-acetyl-D-(13C6)gluco-
pyranose (1) (4.05 g, 10.2 mmol) in DMF (10 mL) was
treated with hydrazine acetate (1.13 g, 12.3 mmol) and
stirred under nitrogen for 2 h. A second portion of
hydrazine acetate (470 mg, 5.11 mmol) was added and
stirred for a further 2 h. EtOAc (50 mL) was then added
followed by water (50 mL) and the organic layer was
washed with water (2 · 50 mL), dried (MgSO4), and
concentrated in vacuo to give 2,3,4,6-tetra-O-acetyl-D-
1
(13C6)glucopyranose (2) as a clear oil (3.28 g, 91%). H

1746
Z. Dinev et al. / Carbohydrate Research 341 (2006) 1743–1747
NMR (400 MHz, CDCl3): d 2.02–2.10 (m, 12H, 4 · CH3),
combined fractions were pooled, concentrated in vacuo
and purified a second time by flash chromatography
(10–25% H2O in acetonitrile) to give uridine 50-diphos-
3.57–5.80 (m, 7H, H-1,2,3,4,5,6a,6b).
1.2.3. Diphenyl 2,3,4,6-tetra-O-acetyl-a-D-(13C6)gluco-
pyranosyl phosphate (3). DMAP (2.20 g, 18.0 mmol)
was added to a solution of 2,3,4,6-tetra-O-acetyl-D-
(13C6)glucopyranose (2) (2.66 g, 7.51 mmol) in CH2Cl2
(53 mL) and stirred at rt for 15 min. The reaction mix-
ture was cooled to ꢀ10 ꢁC, then diphenyl chlorophos-
phate (3.75 mL, 18.1 mmol) was added dropwise. After
stirring for 1 h, the reaction mixture was diluted with
CH2Cl2 (50 mL) and washed successively with ice cold
water (2 · 100 mL), ice cold 0.5 M HCl (2 · 100 mL),
and satd aq NaHCO3 (2 · 100 mL). The organic frac-
tion was dried (MgSO4), concentrated in vacuo, and
purified by flash chromatography (40–50% EtOAc in
petroleum spirits) to give diphenyl 2,3,4,6-tetra-O-acet-
yl-a-D-(13C6)glucopyranosyl phosphate (3) as a pale
pho-2,3,4,6-tetra-O-acetyl-a-D-(13C6)glucopyranose
(5)
(2.14 g, 84%) as a white solid. 1H NMR (400 MHz,
D2O): d 1.88 (s, 3H, CH3), 1.91 (s, 3H, CH3), 1.94 (s,
3H, CH3), 1.97 (s, 3H, CH3), 3.75–5.53 (m, 7H, H-
1,2,3,4,5,6a,6b), 3.99–4.19 (m, 4H, H-20,30,40,50), 5.80
(d, J1 ;2 ¼ 4:0 Hz, 1H, H-10), 5.83 (d, J5 ;6 ¼ 8:0 Hz,
0
0
00 00
1H, H-500), 7.84 (d, J5 ;6 ¼ 8:0 Hz, 1H, H-600); 13C
00 00
NMR (100.5 MHz, D2O): d 20.24, 20.29, 20.30, 20.40
(4C, CH3), 61.51–62.01 (m, 1C, C-6), 65.13 (d,
JC5 ;P ¼ 5:3 Hz, C-50), 67.55–68.95 (m, 2C, H-4,5),
0
69.50–71.80 (m, 3C, C-2,3,30), 74.18 (C-20), 83.10 (d,
JC4 ;P ¼ 9:1 Hz, C-40), 88.82 (C-10), 92.20–92.69 (m, 1C,
0
C-1), 102.84 (C-500), 141.97 (C-600), 151.88 (C-200),
166.31 (C-400), 172.97 (·2), 173.25, 173.83 (4C, C@O);
31P NMR (161.8 MHz, D2O): d ꢀ11.74, ꢀ9.47 (2 · br
s, P-1,2); ESIMS m/z calcd for 13C6C17H32N2O21P2
[MꢀH]ꢀ: 739.1099. Found 739.1114. [M+Naꢀ2H]ꢀ:
761.0929. Found 761.0926.
1
yellow oil (2.70 g, 83%). H NMR (400 MHz, CDCl3):
d 1.81 (s, 3H, CH3), 1.99 (s, 3H, CH3), 2.00 (s, 6H,
CH3 · 2), 3.84 (dd, JC,H = 150 Hz, J6a,6b = 12.8 Hz,
1H, H-6b), 4.03 (dm, JC,H = 148 Hz, 1H, H-5), 4.15
(ddd, JC,H = 147 Hz, J6a,6b = 12.8 Hz, J5,6a = 3.6 Hz,
1H, H-6a), 5.00 (dm, JC,H = 146 Hz, 1H, H-2), 4.88–
5.72 (m, 2H, H-3,4), 6.04 (dm, JC,H = 180 Hz, 1H, H-
1); 13C NMR (100.5 MHz, CDCl3): d 20.21, 20.51,
20.58 (·2) (4C, CH3), 60.93 (d, JC,C = 44.2 Hz, 1C, C-
1.4. Bis(ammonium) uridine 50-diphospho-a-D-(13C6)-
glucose (6)
Uridine 50-diphospho-2,3,4,6-tetra-O-acetyl-a-D-(13C6)-
glucopyranose (5) (1.65 g, 2.23 mmol) was suspended
in a mixture of 0.1 M TEAB (60 mL) and CH3OH
(45 mL) and chilled to ꢀ40 ꢁC. Et3N (750 lL,
10.2 mmol) was then added and the reaction mixture
was stored at ꢀ20 ꢁC for 4 days with occasional mixing.
A mixture of 0.1 M TEAB (30 mL) and CH3OH
(22.5 mL) then Et3N (375 lL, 5.1 mmol) were added
and the reaction mixture was stored at ꢀ20 ꢁC for a fur-
ther 4 days. The reaction mixture was then concentrated
in vacuo, keeping the temperature below 30 ꢁC. The
resulting material was passed through size exclusion
resin (Bio-Gel P-2, 3.5 cm · 45 cm) eluted with 250 mM
NH4HCO3. The fractions containing the product were
pooled and lyophilized. The product was passed
through cation exchange resin (Dowex 50W X8-400,
ammonium form, 2 · 5 cm). Fractions containing the
product were pooled and lyophilized to give bis(ammo-
nium) uridine 50-diphospho-a-D-(13C6)glucose (6) as a
6), 68.87–70.09 (m, 4C, C-2,3,4,5), 94.95 (d, JC,C
=
44.2 Hz, 1C, C-1), 120.00–150.24 (4C, Ph), 169.32,
169.76, 169.95, 170.52 (4C, C@O); 31P NMR
(161.8 MHz, CDCl3): d ꢀ14.38 to ꢀ14.22 (m, 1P, P-1);
ESIMS m/z calcd for 13C6C20H29O13P [M+Na]+:
609.1445. Found 609.1445.
1.3. Uridine 50-diphospho-2,3,4,6-tetra-O-acetyl-a-D-
(13C6)glucopyranose (5)
A solution of diphenyl 2,3,4,6-tetra-O-acetyl-a-D-(13C6)-
glucopyranosyl phosphate (3) (2.01 g, 3.42 mmol) in
EtOAc/EtOH (1:1, 60 mL) containing PtO2 (486 mg)
was shaken under H2 in a Parr apparatus at 380 kPa
for 18 h. The reaction mixture was filtered through Cel-
ite, and the filtrate treated with Et3N (0.95 mL,
6.84 mmol), and then concentrated in vacuo to yield
crude bis(triethylammonium) 2,3,4,6-tetra-O-acetyl-a-
D-glucopyranosyl phosphate (4). Without purification,
this crude material was co-evaporated with dry pyridine
(3 · 20 mL). UMP-morpholidate (3.38 g, 4.91 mmol)
was co-evaporated with pyridine (3 · 20 mL), then trans-
ferred via a cannula into the reaction flask. The com-
bined reagents were co-evaporated with pyridine
(2 · 20 mL) then 1H-tetrazole (772 mg, 11.0 mmol) was
added and the reaction mixture stirred at rt for 24 h.
The reaction mixture was concentrated in vacuo and
the resulting material was purified by flash chromato-
graphy (7:2:1 then 5:2:1, EtOAc/CH3OH/H2O). The
1
white solid (1.28 g, 94%). H NMR (400 MHz, D2O):
d 3.05–4.15 (m, 9H, H-2,3,30,4,40,5,50,6a,6b), 4.19 (d,
J1 ;2 ¼ 3:6 Hz, 1H, H-20), 5.43 (dm, JC,H = 173 Hz,
0
0
1H, H-1), 5.74–5.80 (m, 2H, H-10,500), 7.77 (d, J5 ;6
¼
00 00
8:4 Hz, 1H, H-600); 13C NMR (100.5 MHz, D2O): d
0
60.99 (d, JC,C = 42.7 Hz, 1C, C-6), 65.63 (d, JC5 ;P
¼
5:3 Hz, 1C, C-50), 70.02 (ddd, JC3,C4 = 37.4 Hz,
JC4,C5 = 37.4 Hz, JC2,C4 = 3.0 Hz, 1C, C-4), 70.34 (s,
1C, C-30), 71.79–72.74 (m, 1C, C-2), 74.97 (m, 3C, C-
20,3,5), 83.93 (d, JC4 ;P ¼ 9:1 Hz, 1C, C-40), 89.08 (1C,
0
C-10), 96.29 (dm, JC1,C2 = 44.2 Hz, 1C, C-1), 103.37
(C-500), 142.33 (C-600), 152.56 (C-200), 166.98 (C-400); 31P

Z. Dinev et al. / Carbohydrate Research 341 (2006) 1743–1747
1747
5. Fairweather, J. K.; Him, J. L. K.; Heux, L.; Driguez, H.;
Bulone, V. Glycobiology 2004, 14, 775–781.
6. Wong, C.-H.; Halcomb, R. L.; Ichikawa, Y.; Kajimoto, T.
Angew. Chem., Int. Ed. Engl. 1995, 34, 521–546.
7. Ma, X. Y.; Sto¨ckigt, J. Carbohydr. Res. 2001, 333, 159–
163.
8. Salam, M. A.; Behrman, E. J. Carbohydr. Res. 1982, 101,
339–342.
9. Ernst, C.; Klaffke, W. J. Org. Chem. 2003, 68, 5780–5783.
10. Arlt, M.; Hindsgaul, O. J. Org. Chem. 1995, 60, 14–15.
11. Hanessian, S.; Lu, P. P.; Ishida, H. J. Am. Chem. Soc.
1998, 120, 13296–13300.
NMR (161.8 MHz, D2O): ꢀ10.91 (ddd, JP1,P2 = 19.8 Hz,
JC1,P2 6.9 Hz, JC2,P2 = 6.1 Hz, 1P, P-1), ꢀ9.28 (d,
JP1,P2 = 19.8 Hz, 1P, P-2); LRMS (ESIꢀ, m/z): [MꢀH]ꢀ
571.2, [Mꢀ2H]2ꢀ 285.2. ESIMS m/z calcd for 13C6C9-
H24N2O17P2 [MꢀH]ꢀ: 571.0673. Found 571.0665.
[M+Na-2H]ꢀ 593.0492. Found 593.0494. Anal. Calcd
for 13C6C9H30N4O17P2: C, 30.70; H, 4.99; N, 9.24.
Found: C, 30.67; H, 5.02; N, 9.31.
12. Moffatt, J. G.; Khorana, H. G. J. Am. Chem. Soc. 1961,
83, 649–658.
Acknowledgments
13. Moffatt, J. G.; Khorana, H. G. J. Am. Chem. Soc. 1958,
80, 3756–3761.
14. Wittmann, V.; Wong, C. H. J. Org. Chem. 1997, 62, 2144–
2147.
This work was supported by Biosupplies Australia Pty
Ltd. Thomas Munro is thanked for helpful advice.
15. Oehrlein, R.; Baisch, G. Process for the complete removal
of protective groups on nucleoside diphosphate and
triphosphate sugars, WO 96/24683, 1996.
16. Zamyatina, A.; Gronow, S.; Puchberger, M.; Graziani, A.;
Hofinger, A.; Kosma, P. Carbohydr. Res. 2003, 338, 2571–
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17. Zamyatina, A.; Gronow, S.; Oertelt, C.; Puchberger, M.;
Brade, H.; Kosma, P. Angew. Chem., Int. Ed. 2000, 39,
4150–4153.
Supplementary data
Supplementary data associated with this article can be
found, in the online version, at doi:10.1016/j.carres.
2006.02.033.
18. Graziani, A.; Zamyatina, A.; Kosma, P. Carbohydr. Res.
2004, 339, 147–151.
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