F. Charmantray, P. Dellis, V. Hélaine, S. Samreth, L. Hecquet
FULL PAPER
phases were washed with saturated aq NaHCO3 (20 mL) and water
(20 mL) and dried with MgSO4. After evaporation under vacuum,
a yellow oil was obtained in 95% yield (5.8 g). 1H NMR (400 MHz,
CDCl3): δ = 1.17 (td, J = 7 Hz, 6 H, 2ϫCH3), 1.57 (t, J = 8 Hz,
1 H, SH), 2.6 (ddd, J = 7, 8, 14 Hz, 1 H, 3-H), 2.64 (s, 1 H, OH),
2.75 (ddd, J = 4, 8, 14 Hz, 1 H, 3-H), 3.55 (qd, J = 7, 10 Hz, 2 H,
CH2), 3.62 (ddd, J = 4, 6, 7 Hz, 1 H, 2-H), 3.72 (qd, J = 7, 10 Hz, 1
H, CH2O), 4.43 (d, J = 6 Hz, 1 H, 1-H) ppm. 13C NMR (100 MHz,
CDCl3): δ = 15.3 (CH3), 26.7 (C-3), 63.5 (CH2), 63.8 (CH2), 72.4
(C-2), 103.5 (C-1) ppm.
α Isomer: 1H NMR (400 MHz, CD3OD): δ = 2.94 (dd, J = 5,
11 Hz, 1 H, 5-H), 3.03 (dd, J = 5, 11 Hz, 1 H, 5Ј-H), 3.73 (d, J =
11 Hz, 1 H, 1-H), 3.81 (d, J = 11 Hz, 1 H, 1Ј-H), 4.03 (d, J = 5 Hz,
1 H, 3-H), 4.23–4.32 (m, 1 H, 4-H) ppm. 13C NMR (100 MHz,
CD3OD): δ = 35.9 (C-5), 66.8 (C-1), 79.0 (C-4), 83.9 (C-3), 96.0
(C-2) ppm. HR-ESI-MS calculated for C5H10O4NaS [M + Na]+:
189.0198; found 189.0192.
5-Chloro-D-xylulose (7): 2-Chloroacetaldehyde (850 µL, mmol) at
50% (w/v) in water was added to an aqueous solution of DHAP
(60 mL, 5.7 mmol) at pH 6.8. FruA (200 U) was added. After 1 h
an additional amount of 2-chloroacetaldehyde (150 µL) was added
to the reaction mixture and allowed to react for 18 h at room tem-
perature. The pH of the water phase was adjusted to 4.7 with HCl
(1 ), and acid phosphatase (50 U) was added. The reaction mix-
ture was shaken for a further 48 h at room temperature. After com-
pletion of the reaction, the pH was raised to 7.0 with NaOH (1 ),
and MeOH (180 mL) was poured into the solution. The resulting
precipitate was removed by filtration through celite. The filtrate was
concentrated under vacuum. Column chromatography (CH2Cl2/
MeOH, 9:1) yielded 5-chloro--xylulose as a pale yellow oil
(679 mg, 71% from DHAP). 1H NMR (400 MHz, CD3OD): δ =
3.54 (dd, J = 8, 10 Hz, 1 H, 5-H), 3.70 (dd, J = 8, 10 Hz, 1 H, 5Ј-
H), 4.10 (td, J = 2, 8 Hz, 1 H, 4-H), 4.39 (d, J = 2 Hz, 1 H, 3-H),
4.47 (d, J = 19 Hz, 1 H, 1-H), 4.54 (d, J = 19 Hz, 1 H, 1Ј-H) ppm.
13C NMR (100 MHz, CD3OD): δ = 45.1 (C-5), 67.9 (C-1), 73.6 (C-
4), 76.7 (C-3), 212.5 (C-2) ppm. HR-ESI-MS: calculated for
C5H9ClNaO4 [M + Na]+ 191.0087; found 191.0097.
3-Acetylthioglyceraldehyde Diethyl Acetal (4a): To a solution of po-
tassium thioacetate (962 mg, 8.3 mmol) in thioacetic acid (5.6 mL)
and CH2Cl2 (20 mL) was added dropwise compound 3 (2.23 g,
15.9 mmol) at –78 °C under argon. The mixture was stirred at
–78 °C for 1 h and for 38 h at room temperature. After filtration,
the solution was poured onto ice, and the aqueous phase was ex-
tracted with CH2Cl2 (3ϫ20 mL). The combined organic phases
were washed with saturated aq NaHCO3 and brine. After drying
with MgSO4 and purifying by column chromatography on silica
gel (cyclohexane/ethyl acetate, 8:2), 2.2 g of a yellow oil was ob-
1
tained (63% yield). H NMR (400 MHz, CDCl3): δ = 1.15 (t, J =
7 Hz, 6 H, CH3), 2.26 (s, 3 H, CH3), 2.65 (s, 1 H, OH), 2.9 (dd, J
= 8, 14 Hz, 1 H, 3-H), 3.18 (dd, J = 4, 14 Hz, 1 H, 3-H), 3.5 (m,
2 H, CH2), 3.67 (m, 3 H, CH2, 2-H), 4.3 (d, J = 5 Hz, 1 H, 1-H)
ppm. 13C NMR (100 MHz, CDCl3): δ = 15.2 (CH3), 30.3 (CH3),
31.2 (C-3), 62.5 (CH2), 63.7 (CH2), 70.9 (C-2), 103.5 (C-1), 195.9
(CO) ppm.
3-Thiophosphateglyceraldehyde Diethyl Acetal (4b): Sodium thio-
phosphate (2.7 g, 6.8 mmol) was added to a solution of compound
3 (1 g, 6.8 mmol) in water (20 mL). The reaction mixture was then
stirred for 4 h at room temperature. After evaporation of water un-
der vacuum, 2.1 g of a white solid was obtained (70% yield). 1H
NMR (400 MHz, D2O): δ = 1.26 (t, J = 7 Hz, 6 H, CH3), 2.86 (dd,
J = 9, 14 Hz, 1 H, 3-H), 3.33 (dd, J = 4, 14 Hz, 1 H, 3-H), 3.73
(m, 2 H, CH2), 3.84 (m, 2 H, CH2, 2-H), 4.6 (d, J = 6 Hz, 1 H, 1-
H) ppm. 13C NMR (100 MHz, D2O): δ = 17.2 (CH3), 34.5 (C-3),
66.8 (CH2), 67.3 (CH2), 74.8 (C-2), 106.5 (C-1) ppm.
5-Bromo-D-xylulose (8): 2-Bromoacetaldehyde (1.3 mL, 2 mmol)
was added to an aqueous solution of DHAP (60 mL, 5.7 mmol) at
pH 6.8. FruA (200 U) was added. After 1 h an additional amount
of 2-bromoacetaldehyde (150 µL) was added to the reaction mix-
ture and allowed to react for 18 h at room temperature. The pH of
the water phase was then adjusted to 4.7 with HCl (1 ) and acid
phosphatase (50 U) was added. The reaction mixture was shaken
for a further 48 h at room temperature. After completion of the
reaction, the pH was raised to 7.0 with NaOH (1 ), and MeOH
(180 mL) was poured into the solution. The resulting precipitate
was removed by filtration through celite. The filtrate was concen-
trated under vacuum. Column chromatography (CH2Cl2/MeOH,
9:1) yielded 5-bromo--xylulose as a pale yellow oil (666 mg, 55%
3-Thioglyceraldehyde (3): An aqueous solution of compound 4 (1 g,
5.54 mmol) in water (32 mL) was treated with Dowex 50WX8 resin
(H+ form, 800 mg) for 24 h. Compound 3 thus obtained (in quanti-
tative yield) was used in the next step without further purification.
1
from DHAP). H NMR (400 MHz, CD3OD): δ = 3.40 (dd, J = 8,
10 Hz, 1 H, 5-H), 3.58 (dd, J = 8, 10 Hz, 1 H, 5Ј-H), 4.15 (td, J =
2, 8 Hz, 1 H, 4-H), 4.44 (d, J = 2 Hz, 1 H, 3-H), 4.51 (d, J = 19 Hz,
1 H, 1-H), 4.54 (d, J = 19 Hz, 1 H, 1Ј-H) ppm. 13C NMR
(100 MHz, CD3OD): δ = 33.4 (C-5), 67.9 (C-1), 73.7 (C-4), 77.0
(C-3), 213.4 (C-2) ppm. HR-ESI-MS: calculated for C5H9BrNaO4
[M + Na]+ 234.9582; found 234.9577.
5-Thio-D-xylulofuranose (2): In a 100 mL flask, Tris buffer (0.2 ,
1.36 g), lithium hydroxypyruvate (25 m, 154 mg), compound 3
(175 m, 16 mL), MgCl2 (3 m, 34 mg) and thiamine pyrophos-
phate (2 m, 52 mg) were added to water (35 mL). The pH of the
solution was adjusted to 7.5 and yeast TK (200 U) was added. The
reaction mixture was stirred at 30 °C under argon until complete
disappearance of α-hydroxypyruvate was detected (Lactate dehy-
drogenase/NADH enzymatic assay). Proteins were precipitated
with methanol (170 mL) and removed by centrifugation at
8000 rpm for 15 min. Dowex 50WX8 resin (H+ form, 10 mL) was
added, the mixture was stirred for 30 min, and the resin was re-
5-Halo-D-xylulofuranose 7 and 8 (One-Pot Reaction from rac-Glyci-
dol): Solid Na2HPO4 (3.71 g, 25 mmol) was added to a solution of
rac-2,3-epoxypropanol (1.9 g, 25 mmol, 50 mL) in distilled water.
The mixture was heated at 100 °C for 3 h and assayed for -glyc-
erol-3-phosphate content. The yield was 61% from (S)-2,3-epoxy-
propanol (30% from rac-2,3-epoxypropanol). To a solution con-
taining -glycerol-3-phosphate (155 m, 10 mL, based on enzy-
matic assay) at pH 6.8 was added GPO/catalase mixture (0.1 mL,
45 U/1800 U), FruA (0.07 mL, 20 U) and aldehyde (2-chloroacetal-
dehyde, 0.13 mL, 2 mmol or 2-bromoacetaldehyde, 545 mg,
–
moved by filtration. Dowex 1X8 resin (HCO3 form, 10 mL) was
added to the filtrate, and the mixture was stirred for 30 min. After
filtration of the resin, the solution was evaporated to dryness, and
crude compound 2 was purified by flash column chromatography
(CH2Cl2/MeOH, 8:2) to afford 110 mg of compound 2 (48% yield).
β Isomer: 1H NMR (400 MHz, CD3OD): δ = 2.57 (dd, J = 9, 10 Hz, 2 mmol). The reaction proceeded with stirring at room temperature
1 H, 5-H), 3.03 (dd, J = 8, 10 Hz, 1 H, 5Ј-H), 3.59 (d, J = 13 Hz, overnight. The pH was adjusted to 4.7 with HCl (1 ) and acidic
1 H, 1-H), 3.62 (d, J = 13 Hz, 1 H, 1Ј-H), 3.73 (d, J = 9 Hz, 1 H, 3- phosphatase (50 U) was added. The reaction mixture was shaken
H), 4.23–4.32 (m, 1 H, 4-H) ppm. 13C NMR (100 MHz, CD3OD): δ for another 24 h at room temperature. The pH was raised to 7.0
= 31.2 (C-5), 67.5 (C-1), 77.0 (C-4), 79.8 (C-3), 89.7 (C-2) ppm.
with NaOH (1 ), and MeOH (30 mL) was poured into the solu-
5530
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Eur. J. Org. Chem. 2006, 5526–5532