A convenient preparation of aldonohydroxamic acids in water and crystal structure of L-erythronohydroxamic acid

Article abstract of DOI:10.1016/S0008-6215(01)00210-5

Hydroxamic acids derived from aldonic acids, namely aldonohydroxamic acids, have become an increasingly important class of inhibitors of enzymes involved in the metabolism of carbohydrates. We now report the straightforward preparation of various types of aldonohydroxamic acids by a new methodology involving the use of commercial 50% aqueous hydroxylamine as the source of the free base hydroxylamine that reacts directly with the corresponding aldonolactone dissolved in water. The reaction proceeds almost instantaneously in water at room temperature, yielding generally pure products in quantitative yield. To date, this methodology is probably the most facile and efficient way to synthesize aldonohydroxamic acids. We also determined by X-ray diffraction analysis the first crystal structure of a free aldonohydroxamic acid reported to date. Crystals of L-erythronohydroxamic acid belonged to the monoclinic system, space group P2₁, a=5.511(3), b=7.556(1), c=8.071(3) A, β=109.10°, and Z=2.

Full text of DOI:10.1016/S0008-6215(01)00210-5

www.elsevier.com/locate/carres
Carbohydrate Research 335 (2001) 195–204
Note
A convenient preparation of aldonohydroxamic acids in
water and crystal structure of
acid
L-erythronohydroxamic
Laurent Salmon,a,* Elise Prost,a Claude Merienne,b Renaud Hardre´,a,†
Georges Morgantc
aLaboratoire de Chimie Bioorganique et Bioinorganique, CNRS-FRE 2127,
Institut de Chimie Mole´culaire d’Orsay (ICMO), Baˆtiment 420, Uni6ersite´ de Paris-Sud, F-91405 Orsay, France
bSer6ice de Radiocristallographie, Institut de Chimie Mole´culaire d’Orsay (ICMO), Uni6ersite´ de Paris-Sud,
F-91405 Orsay, France
cLaboratoire de Cristallochimie Bioinorganique, Faculte´ de Pharmacie, Uni6ersite´ de Paris-Sud,
F-92290 Chaˆtenay-Malabry, France
Received 14 May 2001; received in revised form 8 July 2001; accepted 16 July 2001
Abstract
Hydroxamic acids derived from aldonic acids, namely aldonohydroxamic acids, have become an increasingly
important class of inhibitors of enzymes involved in the metabolism of carbohydrates. We now report the
straightforward preparation of various types of aldonohydroxamic acids by a new methodology involving the use of
commercial 50% aqueous hydroxylamine as the source of the free base hydroxylamine that reacts directly with the
corresponding aldonolactone dissolved in water. The reaction proceeds almost instantaneously in water at room
temperature, yielding generally pure products in quantitative yield. To date, this methodology is probably the most
facile and efficient way to synthesize aldonohydroxamic acids. We also determined by X-ray diffraction analysis the
first crystal structure of a free aldonohydroxamic acid reported to date. Crystals of L-erythronohydroxamic acid
,
belonged to the monoclinic system, space group P21, a=5.511(3), b=7.556(1), c=8.071(3) A, i=109.10°, and
Z=2. © 2001 Elsevier Science Ltd. All rights reserved.
Keywords: Hydroxamic acids; Hydroxylamine; Lactones; Water, reactions in; Monosaccharides; X-ray crystal structure
1. Introduction
first introduced by Mathis more than 50 years
ago with the syntheses of
D
-glucono- (1,
-arabinono-, -man-
-galactonohydroxamic acids.1 Al-
Hydroxamic acids derived from aldonic
acids, namely aldonohydroxamic acids, were
Scheme 1),
nono- and
L
-ribono-,
L
D
D
donohydroxamic acids have become an
increasingly important class of compounds.
They have proven key compounds, as potent
inhibitors, for the study of the mechanistic
and structural aspects of several enzymes in-
volved in the metabolism of simple carbohy-
* Corresponding author. Tel.: +33-169-156311; fax: +33-
169-157281.
E-mail address: lasalmon@icmo.u-psud.fr (L. Salmon).
† Present address: Institute of Fundamental Sciences,
Massey University, Palmerston North, New Zealand.
0008-6215/01/$ - see front matter © 2001 Elsevier Science Ltd. All rights reserved.
PII: S0008-6215(01)00210-5

196
L. Salmon et al. / Carbohydrate Research 335 (2001) 195–204
Scheme 1. Preparation of aldonohydroxamic acids 1–10 from the corresponding aldonolactones 11–20.
allowing removal of some of the precipitated
sodium chloride; or (ii) in aqueous solution
using lithium hydroxide or another base. In
most cases, the aldonohydroxamic acids had
to be further purified by size-exclusion chro-
matography, precipitation, or ion-exchange
chromatography.
In a general project dealing with the study
of aldonohydroxamic acids, we report here the
syntheses of several aldonohydroxamic acids
of various lengths and configurations from
their corresponding aldonolactones: the
known aldonohydroxamic acids 1–4 and new
aldonohydroxamic acids 5–10 (Scheme 1). We
had first prepared these products according to
the classical methodology using hydroxy-
lamine hydrochloride and sodium methoxide
drates, being used as mimetics of the postu-
lated high-energy intermediates or transition
states. The known examples are phosphogly-
colohydroxamic acid,2,3
D
-threonohydroxamic
-arabinonohydroxamic acid 5-
phosphate5 (2, Scheme 1),
-lyxonohydrox-
amic acid6 (3, Scheme 1),
-xylarohydroxa-
mic acid7 and -arabinonohydroxamic acid8
acid,4
D
D
D L
,
D
(4, Scheme 1). These compounds were com-
monly prepared by reaction of the free base
hydroxylamine with the corresponding methyl
or ethyl aldonates or aldonolactones. How-
ever, and as far as the experimental details
were reported, the free base hydroxylamine
was always obtained by neutralization of a
hydroxylamine hydrochloride solution either:
(i) in dry methanol using sodium methoxide,

L. Salmon et al. / Carbohydrate Research 335 (2001) 195–204
197
in methanol. A large amount of sodium chlo-
ride was always present with the product,
which proved very difficult to remove com-
pletely without hydrolysis of the aldonohy-
droxamic acid to the corresponding aldonate
during the purification steps, such as during
size-exclusion chromatography. We observed
that most of the prepared aldonohydroxamic
acids are more or less unstable in water at
room temperature, even at neutral pH. Hence,
we report in this paper, the synthesis of the
pure analytical aldonohydroxamic acids 1–10
by a new methodology which does not involve
any chromatographic purification step: the use
of commercial 50% aqueous hydroxylamine as
the source of the free base hydroxylamine that
reacts directly with the corresponding aldono-
lactone dissolved in water. There are only a
few examples in the literature that reported
the use of 50% aqueous hydroxylamine, but as
far as we know, only for the synthesis of
hydroxamic acid derivatives of amino acids (in
ethanol or THF).9,10 For one of the prepared
are commercially available (Aldrich, Acros).
-Arabinono-1,4-lactone 5-phosphate (12)14
and
-lyxono-1,4-lactone (17)15 were prepared
as previously described. -Lyxono-1,4-lactone
(13),16,17 -arabinono-1,4-lactone (14),18,19
erythrono-1,4-lactone (15)20 and
-threono-
1,4-lactone (16)21 were obtained from barium
-lyxonate,22 potassium -arabinonate23 (by
hypoiodite oxidation22 of
-arabinose), potas-
sium -threonate
-erythronate23 and calcium
D
L
D
D
L
-
L
D
D
D
L
L
(Aldrich), respectively, using the reported lac-
tonization procedure19 (except that P2O5 was
used instead of magnesium perchlorate). 5-De-
oxy-
L
-lyxono-1,4-lactone 2024,25 was obtained
from
L
-fucose by oxidative alkaline degrada-
tion,26 followed by lactonization.19 New spec-
tral data of the aldonolactones 15, 16, 17 and
20 are reported below.‡
As a general procedure, 50% aqueous hy-
droxylamine (Aldrich, 306 mL, 5 mmol) was
added to a solution of the lactone (1 mmol) in
water (1.7 mL). The reaction mixture was then
stirred for 10 min at room temperature. The
corresponding aldonohydroxamic acid was
obtained by evaporation of water and hydrox-
ylamine under diminished pressure (T=25
°C, P=0.1 torr). Complete removal of hy-
droxylamine was achieved by adding water
(5–10 mL) to the residue, followed by evapo-
ration; this procedure was repeated until a
constant mass was reached. Generally, the
final lyophilization step produced the pure
product in quantitative yield. 13C NMR analy-
sis in deuterium oxide at neutral pH showed
the C-1 chemical shift at approximately 171.0
ppm, a value characteristic of aldonohydrox-
amic acids. Possible hydrolysis of the product
to the corresponding hydroxylammonium al-
donate would give a typical C-1 resonance at
179–180 ppm. As a matter of fact, this was
observed in the case of compounds 1, 3, 5 and
7; while these compounds were at least 95%
pure according to their NMR spectra, they
had to be recrystallized to give pure analytical
products. All aldonohydroxamic acids re-
ported in this paper gave spectroscopic data
products,
L
-erythronohydroxamic acid 5, we
were able to obtain crystals suitable for X-ray
diffraction analysis (attempts to crystallize the
other analogs either failed or led to unsuitable
crystals). Crystal structures for two aldonohy-
droxamic acids,
D
-threonohydroxamic acid
and phosphoglycolohydroxamic acid, com-
plexed at the active site of their target enzymes
were reported.4,11–13 We now report the first
crystal structure of a free aldonohydroxamic
acid. It will contribute to the better under-
standing of the solid-state structure of
monosaccharides derivatives, and more partic-
ularly of aldonohydroxamic acid derivatives.
2. Results and discussion
Synthesis.—Aldonohydroxamic acids 1–10
were prepared from their corresponding al-
donolactones,
D
-glucono-1,5-lactone (11), and
the aldono-1,4-lactones of the following
configurations: 5-phospho-
D
-arabino- (12),
D
-
lyxo- (13),
D
-arabino- (14),
-lyxo- (17),
-ribo- (19) and 5-deoxy-
-Glucono-1,5-lactone (11),
1,4-lactone (18) and -ribono-1,4-lactone (19)
L
-erythro- (15),
‡ 13C NMR (D2O, 50.3 MHz): 15: l 179.4 (C-1); 73.8 (C-4);
70.5 (C-2); 69.6 (C-3). 16: l 178.0 (C-1); 73.9 (C-2); 73.0
(C-3); 70.3 (C-4). 17: l 179.0 (C-1); 82.4 (C-4); 71.3 (C-2);
70.3 (C-3); 60.6 (C-5). 20: l 179.5 (C-1); 79.3 (C-4); 72.0, 71.8
(C-2, C-3); 13.8 (C-5).
L
-threo- (16),
L
D
-erythro- (18),
D
L
-lyxo- (20) (Scheme
-erythrono-
1).
D
D
D

198
L. Salmon et al. / Carbohydrate Research 335 (2001) 195–204
Table 2
and microanalyses in full agreement with the
proposed structures (see Section 3).
Fractional atomic coordinates and equivalent isotropic ther-
mal parameter Ueq for non-hydrogen atoms of
L
-erythrono-
Our methodology offers the obvious advan-
tage that there is no salt to remove. The
reaction proceeds almost instantaneously in
water at room temperature, yielding, in most
cases, pure products in quantitative yield. Ex-
cept for a few of the prepared aldonohydrox-
amic acids, there are no side-products, no
work-up and no purification step. There is no
hydroxamic acid 5 a
b
Atom
x/a
y/b
z/c
Ueq
O-1
O-2
O-3
O-4
O-5
N-5
C-1
C-2
C-3
C-4
−0.0496(4)
0.4719(4)
0.0049(5)
0.4894(5)
0.7521(4)
0.5265(4)
0.0614(3) 0.4248(3)
0.0663(3) 0.4396(2)
0.1771(3) 0.0169(3)
0.2294(3) 0.0182(3)
0.4951(3) 0.2228(2)
0.4240(3) 0.2372(3)
0.0339
0.0294
0.0337
0.0292
0.0264
0.0235
0.0315
0.0237
0.0222
0.0210
0.0496(6) −0.0514(4) 0.3191(4)
Table 1
0.2704(5)
0.1901(5)
0.4163(5)
0.0336(3) 0.2784(3)
0.2090(3) 0.1796(3)
0.2888(3) 0.1378(3)
Crystal parameters, data collection and structure refinement
for L-erythronohydroxamic acid 5
Crystal parameters
Molecular formula
Molecular weight
Crystal system
a E.s.d’s in parentheses refer to the last significant digit. Ueq
is defined as the cube root of the product of the principal
C4H9NO5
151.12
monoclinic
P21
axes.
b 100% occupancy.
Space group
Unit cell dimensions
need to synthesize a hydroxylamine reagent,
and no need for protective groups on the
starting aldonolactones. To date, this method-
ology using 50% aqueous hydroxylamine is
probably the easiest and most efficient way to
synthesize aldonohydroxamic acids. There is
no doubt that it will soon prove useful in the
synthesis of other types of hydrophilic hydrox-
amic acid derivatives.
,
a (A)
5.511(3)
7.556(1)
8.071(3)
109.10
317.6(2)
2
1.58
0.14
,
b (A)
,
c (A)
i (°)
3
,
V (A )
Z
zcalcd (g cm−3
)
v (Mo Ka) (cm−1
F(000)
)
160.08
0.25×0.35×0.45
colorless hexagonal rods
Crystal size (mm)
Crystal color and shape
Determination of the X-ray structure of
L
-
erythronohydroxamic acid 5.—Crystal struc-
ture of 5 contributes to the study of the
solid-state structures of monosaccharides
aimed at a better understanding of the factors
that determine which structure type is formed,
particularly in the case of aldonohydroxamic
acids for which, to our knowledge, no crystal
structure has been reported to date. The
refined unit cell constants and other relevant
Data collection
Diffractometer
Monochromator
Radiation
Scan mode
Temperature (K)
q Range for data collection (°)
Index range of h, k, l
Enraf–Nonius CAD4
graphite
Mo Ka (u=0.71069 A)
ꢀ−2q
291
,
2–30
−85h58,
05k512,
05l513
DIFABS
0.82
Absorption correction
Tmin
crystal data for -erythronohydroxamic acid 5
L
are presented in Table 1, together with details
of the intensity measurements. Final fractional
atomic coordinates and equivalent isotropic
temperature factors for non-hydrogen atoms
of compound 5 are reported in Table 2. Se-
lected torsion angles and hydrogen-bond
parameters are illustrated in Tables 3 and 4,
respectively. The ORTEP view of the title com-
pound is represented in Fig. 1, together with
the labeling of the atoms and hydrogen bonds.
The molecular packing scheme is represented
in Fig. 2.
Tmax
1.00
2945/1469
853
Reflections collected/unique
Reflections observed [I\3|(I)]
Structure refinement
Parameters
Final R
95
0.0388
0.0422
Chebyshev
1.07, 1.18, −0.193
1.0339
0.011643
−0.215/0.224
Final Rw
Weighting scheme
Coefficient Ar
Goodness-of-fit on F2
(Z/|)max
Dzmin/Dzmax (e A
−3
,
)

L. Salmon et al. / Carbohydrate Research 335 (2001) 195–204
199
Table 3
group is not in the linear extension of the
zigzag chain, but adopts a gauche orientation
(Fig. 1), as observed in the crystal structure of
Selected torsion angles for atoms of
acid 5
L
-erythronohydroxamic
various compounds like
L
-mannonic acid
Sequence
Angle (°)
hydrazide.30
O-1ꢀC-1ꢀC-2ꢀC-3
C-1ꢀC-2ꢀC-3ꢀC-4
O-2ꢀC-2ꢀC-3ꢀC-4
O-3ꢀC-3ꢀC-4ꢀN-5
C-2ꢀC-3ꢀC-4ꢀN-5
O-4ꢀC-4ꢀN-5ꢀO-5
C-3ꢀC-4ꢀN-5ꢀO-5
C-4ꢀN-5ꢀO-5ꢀH-5
−59.40
−178.89
61.18
136.68
−104.42
−6.82
173.16
90.05
Figs. 1 and 2 show that the hydroxamic
acid moiety of compound 5 is in the Z confor-
mation. The O-4ꢀC-4ꢀN-5ꢀO-5 torsion angle
is −6.8°, showing that the hydroxamic acid
moiety is not quite planar, probably because it
forms an extensive hydrogen-bonding network
with neighboring molecules. This type of con-
formation was observed in the solid state for
several hydroxamic acids like cyclohexanecar-
bohydroxamic acid31 and formohydroxamic
acid.27 Bond lengths and angles of the hydrox-
amic acid moiety of 5 are close to the mean
values reported for a series of N-unsubstituted
hydroxamic acids.27 The H-5ꢀO-5 bond is
turned out of the plane of the hydroxamic
acid group (C-4ꢀN-5ꢀO-5ꢀH-5 torsion angle is
90°), in agreement with the reported crystal
structures of various hydroxamic acids which
show HꢀOꢀNꢀC torsion angles in the range
9(50–140)°.27,28,31 This orientation of the
NꢀOH hydroxyl group is obviously more ap-
propriate for intermolecular hydrogen bond-
ing, and appears as an important factor
determining the conformation in crystals of
hydroxamic acid derivatives. In contrast, ab
initio studies on hydroxamic acids generally
show the NꢀOH hydroxyl group intramolecu-
larly hydrogen-bonded to the oxygen of the
carbonyl group, which puts the NꢀOH hy-
droxyl group in the plane of the hydroxamic
acid moiety.32–34
Crystals of
L
-erythronohydroxamic acid 5
belonged to the monoclinic space group P21
,
with a=5.511(3), b=7.556(1), c=8.071(3) A,
i=109.10°. The unit cell consists of two
molecules of
L
-erythronohydroxamic acid 5
(Fig. 2). X-ray crystallography revealed that
compound 5 crystallizes without solvent
molecules, as observed in the case of formohy-
droxamic acid,27 but contrarily to acetohy-
droxamic acid which was reported to
crystallize as its hemihydrate.28
Although comparison with other aldonohy-
droxamic acids is not possible, bond lengths
and angles for the polyhydroxylated chain of
compound 5 all compare well with the corre-
sponding average values found from X-ray
studies of alditols, like
D
-glucitol for exam-
ple.29 The same holds for torsion angles (Table
3). The carbon chain adopts an extended pla-
nar zigzag conformation. As observed for
tetritols and some other alditols,29 no unfavor-
able 1,3-parallel OꢀꢀO or CꢀꢀO interactions (as
well as no intramolecular hydrogen bonds)
occur in the crystal structure of 5 as a result of
the configuration of the stereogenic centers
C-2 and C-3. However, the terminal hydroxyl
All heteroatoms are involved in a dense
hydrogen-bonding network (Fig. 1). H-Bond
parameters of Table 4 all compare well with
Table 4
Hydrogen bond parameters for L-erythronohydroxamic acid 5
,
Donor
Acceptor
(A)
Distances (A)
Angle (°)
DꢀH···A
Symmetry operation on acceptor
(DꢀH)
D···A
H···A
O-5ꢀH-5
N-5ꢀH-51
O-1ꢀH-1
O-3ꢀH-3
O-2ꢀH-2
O4i
2.645(3)
2.82(3)
2.825(4)
2.818(3)
2.679(3)
1.701(3)
1.876(4)
1.925(3)
2.064(3)
1.793(3)
161.93
159.40
172.43
148.85
162.59
i=−x+1, y+1/2, −z
ii=−x+1, y+1/2, −z+1
iii=−x+1, y−1/2, −z+1
iv=x−1, y, z
O2ii
O5iii
O4iv
O1v
v=x+1, y, z,

200
L. Salmon et al. / Carbohydrate Research 335 (2001) 195–204
Fig. 1. ORTEP view of
L
-erythronohydroxamic acid 5 was generated by the CAMERON program. Atom labeling was generated by
the CRYSTALS program (it does not correspond to the usual labeling of monosaccharides). Thermal ellipsoids are drawn at the
50% level. Dashed lines indicate hydrogen bonds.
Conclusions.—We have developed a novel,
simple and efficient methodology for the syn-
thesis of aldonohydroxamic acids, using com-
mercial 50% aqueous hydroxylamine as the
source of the free base that reacts directly with
the corresponding aldonolactone in water.
The reaction proceeds almost instantaneously
in water at room temperature, yielding, in
most cases, pure products in quantitative
the reported values for various monosaccha-
rides derivatives.35–38 The packing arrange-
ment assumed by -erythronohydroxamic acid
L
5 shows that one molecule interacts with six
other molecules. Molecules of 5 are linked by
‘head-to-tail’ interactions in which the NHꢀO
group of each molecule forms a hydrogen-
bonded bridge across the C-1 and C-2 hy-
droxyl groups of a neighboring molecule,
forming an eight-membered ring in which the
N-5ꢀH-51 and O-1ꢀH-1 groups are H-bond
donors and O-5 and O-2 are H-bond accep-
tors. A similar pattern was recently described
yield. In the case of
L
-erythronohydroxamic
acid, we also report the first crystal structure
of a free aldonohydroxamic acid.
for
D
-mannose oxime.37 Molecules of 5 are
3. Experimental
also linked by ‘head-to-head’ interactions in
which three hydroxamic acid groups are in-
volved: the NꢀOH group of a given molecule
is H-bond donor to the carbonyl oxygen of a
first neighboring molecule, while the oxygen
carbonyl is H-bond acceptor with the NꢀOH
General methods.—Unless otherwise stated,
chemicals (used at their commercial purity)
were obtained from Acros and solvents from
SDS. Melting points were measured on a Re-
ichert apparatus and not corrected. Optical
rotations were measured on a JASCO DIP-
group of
a
second molecule. Finally,
1
molecules of 5 are linked together through
O-2ꢀH-2···O-1 and O-3ꢀH-3···O4 hydrogen
bonds (Fig. 1).
370 digital polarimeter. H NMR spectra were
recorded at 200, 250 or 400 MHz, and were
referenced to residual internal HOD (4.80

L. Salmon et al. / Carbohydrate Research 335 (2001) 195–204
201
ppm). For CH2 groups, H-4%, H-5%, and H-6%
resonances appear arbitrarily at higher field
than H-4, H-5, and H-6 resonances, respec-
tively. 13C NMR spectra were recorded at 50.3
or 62.5 MHz and referenced to internal 1,4-
dioxane (67.40 ppm). Chemical shifts are re-
ported in ppm (l). Infrared spectra were
recorded with a FT-IR Bruker IFS-66 spec-
trometer. Low-resolution mass spectrometry
(MS) and high-resolution mass spectrometry
(HR-MS) analyses of the trimethylsilylated
(TMS) derivatives39 were performed by elec-
trospray with positive ionization mode. Non-
silylated derivatives were analyzed by
low-resolution electrospray mass spectrometry
with negative ionization mode. Evaporations
were performed under diminished pressure be-
low 25 °C. Final products were stored at
−18 °C. Elemental analyses were performed
by the microanalytical laboratory at the ICSN,
CNRS, Gif-sur-Yvette. With aq FeCl3, all al-
donohydroxamic acids had characteristic visi-
ble absorption peaks from u 488 to 498 nm.
Crystal structure analysis.—Crystals of
compound 5 were grown by slow cooling of a
boiling saturated abs EtOH–water solution.
Following filtration, the crystals were rinsed
with abs EtOH and filtered by succion. The
crystals were mounted, using glass fibers, on
an Enraf–Nonius CAD4 diffractometer
equipped with a graphite monochromator.
The lattice parameters were refined using 25
reflections. The data were collected using the
ꢀ−2q scan technique and with Mo Ka radia-
,
tion (u=0.71069 A). During the data collec-
tion, three intensity control reflections were
monitored every 2 h, showing no loss of inten-
sity. The data were corrected for Lorentz and
polarization effects. The structure was solved
by a combination of direct methods using SIR
procedure40 and heavy-atom techniques and
refined by full-matrix least-squares method
based on F2, using the CRYSTALS software.41
An empirical absorption correction with the
42
program DIFABS was used. Anisotropic dis-
placement parameters were assigned to all
non-hydrogen atoms. The hydrogen atoms
were introduced in calculated idealized posi-
,
tions (d(CꢀH)=0.98 A) and their atomic co-
ordinates were recalculated after each cycle.
They were given isotropic thermal parameters
20% higher than those of the carbon to which
they are attached. Least-squares refinements
were performed by minimizing the function
S w(ꢀFoꢀ−ꢀFcꢀ)2, where Fo and Fc are the ob-
served and calculated structure factors. The
weighting scheme used in the last-refinement
cycles was w=w%[1−(DF/6|(Fo)2)]2 where
n
w%=1/S1ArTr(x) with three coefficients Ar for
the Chebyshev polynomial ArTr(x) where x
was Fc/Fc(max).43 Models reached conver-
gence with R=S(ꢀꢀFoꢀ−ꢀFcꢀꢀ)/S(ꢀFoꢀ) and
Rw=[S w(ꢀFoꢀ−ꢀFcꢀ)2/S w(Fo)2]1/2, having val-
ues listed in Table 1. Criteria for a satisfactory
complete analysis were ratios of rms shift to
standard deviation less than 0.1 and no signifi-
cant features in final difference maps. Details
of data collection and refinement are given in
Table 1. Calculations were performed with the
CRYSTALS package program. The drawings of
the
molecules
were
generated
using
44
CAMERON
.
The atomic scattering factors
were taken from International Tables for X-
ray Crystallography.45
D
-Gluconohydroxamic acid 1.—The title
compound was recrystallized from water, abs
EtOH and acetone. Following trituration with
Fig. 2. Molecular packing in L-erythronohydroxamic acid 5.

202
L. Salmon et al. / Carbohydrate Research 335 (2001) 195–204
J2,3 2 Hz, 1 H, H-2); 3.85 (dd, J3,4 9 Hz, 1 H,
H-3); 3.71 (m, 1 H, H-4); 3.82 (dd, J4,5 2 Hz,
1 H, H-5); 3.63 (dd, J4,5% 6, J5,5% −11 Hz, 1 H,
H-5%). 13C NMR (D2O, 62.5 MHz): l 172.9
(C-1); 72.5, 71.7, 71.6 (C-2, C-3, C-4); 64.2
(C-5). MS (negative mode): 180.3 [M−H]−.
Anal. Calcd for C5H11NO6: C, 33.15; H, 6.12;
N, 7.73; Found: C, 32.81; H, 5.93; N, 7.77.
cold abs EtOH, then with Et2O, it was finally
dried under diminished pressure to give a
white powder, mp 140–143 °C (lit.1 138–
140 °C), yield: 110 mg (75%), [h]2D1 +20.8° (c
1.0, water). IR (KBr) w 1654 (CO); 3221, 3445
1
(NH, OH) cm−1. H NMR (D2O, 400 MHz):
l 4.38 (d, J2,3 5 Hz, 1 H, H-2); 4.12 (dd, J3,4
3
Hz, 1 H, H-3); 3.86 (dd, J5,6 3, J6,6% −12 Hz,
1 H, H-6); 3.79 (ddd, J4,5 8 Hz, 1 H, H-5); 3.73
(dd, 1 H, H-4); 3.69 (dd, J5,6% 6 Hz, 1 H, H-6%).
13C NMR (D2O, 50.3 MHz): l 171.1 (C-1);
73.3, 72.4, 71.9, 71.1 (C-2, C-3, C-4, C-5); 63.5
(C-6). MS (negative mode): 210.1 [M−H]−.
Anal. Calcd for C6H13NO7: C, 34.13; H, 6.21;
N, 6.63; Found: C, 33.91; H, 6.14; N, 6.48.
L
-Erythronohydroxamic acid 5.—The title
compound was recrystallized from hot abs
EtOH and water to give colorless crystals, mp
131–132.5 °C, yield: 76 mg (40%), [h]D22
−
12.9° (c 1.0, water). IR (KBr) w 1661 (CO);
1
3318 (br, NH, OH) cm−1. H NMR (D2O,
250 MHz): l 4.15 (d, J2,3 5 Hz, 1 H, H-2); 3.89
(m, 1 H, H-3); 3.67 (dd, J3,4 4, J4,4% −12 Hz,
D
-Arabinonohydroxamic acid 5-phosphate,
13
1 H, H-4); 3.60 (dd, J3,4% 7 Hz, 1 H, H-4%). C
disodium salt (2).—The title compound was
obtained from its hydroxylammonium precur-
sor by elution on a Dowex® 50X4-400 column
(Na+ form, water). Following evaporation,
then lyophilization, the title compound was
obtained as a white fluffy powder, yield: 106
NMR (D2O, 50.3 MHz): l 171.2 (C-1); 73.3,
72.0 (C-2, C-3); 62.5 (C-4). MS (negative
mode): 150.0 [M−H]−. Anal. Calcd for
C4H9NO5: C, 31.79; H, 6.00; N, 9.27; Found:
C, 31.95; H, 5.79; N, 9.06.
L
-Threonohydroxamic acid 6.—Lyophiliza-
1
mg (90%). IR, H and 13C NMR data are in
tion of the title compound gave a white fluffy
powder, mp 137–138.5 °C, yield: 471 mg
(100%), [h]2D4 +46.9° (c 1.0, water). IR (KBr)
w 1636, 1670 (CO); 3184, 3266 (br, NH, OH)
agreement with the literature.5,14 Anal. Calcd
for C5H10NNa2O9P·2.5 H2O: C, 17.15; H,
4.32; N, 4.00; Na, 13.13; P, 8.85; Found: C,
16.90; H, 3.98; N, 3.69; Na, 13.30; P, 8.82.
1
cm−1. H NMR (D2O, 200 MHz): l 4.24 (d,
D
-Lyxonohydroxamic acid 3.—The title
J2,3 3 Hz, 1 H, H-2); 3.98 (ddd, J3,4 6, J3,4%
7
compound was recrystallized from abs EtOH
and water, then lyophilized to give a white
fluffy powder, mp 147.5–149 °C, yield: 168
mg (33%), [h]2D1 +2.5° (c 1.0, water). IR
(KBr) w 1661 (CO); 3261, 3371, 3468 (NH,
Hz, 1 H, H-3); 3.66 (dd, J4,4% −12 Hz, 1 H,
H-4); 3.59 (dd, 1 H, H-4%). 13C NMR (D2O,
50.3 MHz): l 171.8 (C-1); 72.6, 71.6 (C-2,
C-3); 62.9 (C-4). MS (TMS deriv.): 534.2
[C19H49NO5Si5+Na]+, 462.1 [C16H41NO5-
Si4+Na]+, 440.2, 390.1, 285.1. Anal. Calcd
for C4H9NO5: C, 31.79; H, 6.00; N, 9.27;
Found: C, 31.56; H, 5.93; N, 9.06.
1
OH) cm−1. H NMR (D2O, 250 MHz): l 4.13
(d, J2,3 7 Hz, 1 H, H-2); 3.87 (ddd, J3,4 2, J4,5
5, J4,5% 8 Hz, 1 H, H-4); 3.77 (dd, 1 H, H-3);
3.65 (dd, J5,5% −12 Hz, 1 H, H-5); 3.60 (dd, 1
L
-Lyxonohydroxamic acid 7.—The title
13
H, H-5%). C NMR (D2O, 50.3 MHz): l 171.6
compound was recrystallized from abs EtOH
and water, then lyophilized to give a white
fluffy powder, mp 147.5–148 °C, yield: 137
mg (56%), [h]2D1 −2.6° (c 1.0, water). IR
(KBr) w 1661 (CO); 3265, 3371, 3469 (NH,
(C-1); 71.6, 71.3, 71.0 (C-2, C-3, C-4); 63.5
(C-4). MS (TMS deriv.): 420.2 [C14H35-
NO6Si3+Na]+, 398.2 [C14H35NO6Si3+H]+,
348.1. Anal. Calcd for C5H11NO6: C, 33.15; H,
6.12; N, 7.73; Found: C, 33.07; H, 6.04; N,
7.58.
1
OH) cm−1. H NMR (D2O, 250 MHz): l 4.14
(d, J2,3 7 Hz, 1 H, H-2); 3.88 (ddd, J3,4 2, J4,5
5, J4,5% 8 Hz, 1 H, H-4); 3.78 (dd, 1 H, H-3);
3.66 (dd, J5,5% −12 Hz, 1 H, H-5); 3.60 (dd, 1
D
-Arabinonohydroxamic
acid
4.—
Lyophilization of the title compound gave a
13
white fluffy powder, mp 146–148 °C ( -ara-
L
H, H-5%). C NMR (D2O, 50.3 MHz): l 171.6
binonohydroxamic acid: lit.1 150 °C), yield:
500 mg (99%), [h]2D0 −41.1° (c 1.0, water). IR
(KBr) w 1629, 1665 (CO); 3281 (br NH, OH)
(C-1); 71.6, 71.3, 71.0 (C-2, C-3, C-4); 63.5
(C-4). MS (negative mode): 180.0 [M−H]−.
Anal. Calcd for C5H11NO6: C, 33.15; H, 6.12;
N, 7.73; Found: C, 33.07; H, 5.89; N, 7.74.
cm−1. H NMR (D2O, 250 MHz): l 4.47 (d,
1

L. Salmon et al. / Carbohydrate Research 335 (2001) 195–204
203
4. Supplementary material
D
-Erythronohydroxamic acid 8.—The title
compound was triturated with abs EtOH, and
then with Et2O. It was finally dried under
diminished pressure to give a pale yellow pow-
der, mp 129–130.5 °C, yield: 188 mg (99%),
[h]2D4 +14.1° (c 1.0, water). IR (KBr) w 1629,
Full crystallographic details, excluding
structure features, have been deposited with
the Cambridge Crystallographic Data Center
(deposition no. CCDC 166786). These data
may be obtained, on request, from The Direc-
tor, CCDC, 12 Union Road, Cambridge CB2
1EZ, UK (Fax: +44-1223-336033, e-mail: de-
posit@ccdc.cam.ac.uk or www: http://
www.ccdc.cam.ac.uk).
1661 (CO); 3226, 3386, 3432 (NH, OH) cm−1
.
5
1H NMR (D2O, 200 MHz): l 4.16 (d, J2,3
Hz, 1 H, H-2); 3.90 (ddd, J3,4 4, J3,4% 7 Hz, 1
H, H-3); 3.69 (dd, J4,4% −12 Hz, 1 H, H-4);
3.61 (dd, 1 H, H-4%). 13C NMR (D2O, 50.3
MHz): l 171.0 (C-1); 73.2, 72.0 (C-2, C-3);
62.5 (C-4). MS (TMS deriv.): 462.2
[C16H41NO5Si4 +Na]+, 440.2 [C16H41NO5-
References
Si4+H]+,
390.1
[C13H33NO5Si3 +Na]+,
1. Mathis, F. C. R. Acad. Sci. Paris 1949, 229, 226–227.
2. Collins, K. D. J. Biol. Chem. 1973, 249, 136–142.
3. Lewis, D. J.; Lowe, G. J. Chem. Soc., Chem. Commun.
1973, 713–715.
4. Allen, K. N.; Lavie, A.; Petsko, G. A.; Ringe, D. Bio-
chemistry 1995, 34, 3742–3749.
5. Hardre´, R.; Bonnette, C.; Salmon, L.; Gaudemer, A.
Bioorg. Med. Chem. Lett. 1998, 8, 3435–3438.
6. Wieczorek, S. J.; Kalivoda, K. A.; Clifton, J. G.; Ringe,
D.; Petsko, G. A.; Gerlt, J. A. J. Am. Chem. Soc. 1999,
121, 4540–4541.
285.1. Anal. Calcd for C4H9NO5: C, 31.79; H,
6.00; N, 9.27; Found: C, 31.84; H, 5.92; N,
9.31.
D
-Ribonohydroxamic acid 9.—The title
compound was dried under diminished pres-
sure to give a white fluffy powder, mp 133–
135 °C
(
-ribonohydroxamic acid: lit.1
L
133–136 °C), yield: 181 mg (100%), [h]D21
+
16.2° (c 1.0, water). IR (KBr) w 1642 (CO);
7. Gulick, A. M.; Hubbard, B. K.; Gerlt, J. A.; Rayment, I.
Biochemistry 2000, 39, 4590–4602.
8. Kobashi, K.; Kumashi, K.; Hase, J. Biochim. Biophys.
Acta 1971, 227, 429–441.
9. Spengler, J.; Burger, K. J. Chem. Soc., Perkin Trans. 1
1998, 2091–2095.
10. Dankwardt, S. M. Synlett 1998, 761–761.
11. Davenport, R. C.; Bash, P. A.; Seaton, B. A.; Karplus,
M.; Petsko, G. A.; Ringe, D. Biochemistry 1991, 30,
5821–5826.
12. Dreyer, M. K.; Schulz, G. E. J. Mol. Biol. 1996, 259,
458–466.
13. Joerger, A. C.; Gosse, C.; Fessner, W.-D.; Schultz, G. E.
Biochemistry 2000, 39, 6033–6041.
14. Hardre´, R.; Salmon, L. Carbohydr. Res. 1999, 318, 110–
115.
15. Frankel, D. A.; O’Brien, D. F. J. Am. Chem. Soc. 1994,
116, 10057–10069.
1
3170, 3290, 3413 (NH, OH) cm−1. H NMR
(D2O, 250 MHz): l 4.34 (d, J2,3 4 Hz, 1 H,
H-2); 3.90 (dd, J3,4 8 Hz, 1 H, H-3); 3.82 (m,
1 H, H-4); 3.79 (m, J4,5 2 Hz, 1 H, H-5); 3.62
(dd, J4,5% 7, J5,5% −12 Hz, 1 H, H-5%). 13C
NMR (D2O, 50.3 MHz): l 171.0 (C-1); 73.3,
72.5, 71.9 (C-2, C-3, C-4); 63.6 (C-5). MS
(TMS deriv.): 614.3 [C23H59NO6Si6+H]+,
571.7, 204.3, 148.2, 132.2. MS (negative
mode): 180.2 [M−H]−. HRMS: Calcd for
C23H60NO6Si6 (TMS deriv.): 614.3040; Found:
614.3036. Anal. Calcd for C5H11NO6: C,
33.15; H, 6.12; N, 7.73; Found: C, 33.08; H,
6.06; N, 7.69.
16. Thompson, A.; Wolfrom, M. L. J. Am. Chem. Soc. 1946,
68, 1509–1510.
17. Angelotti, T.; Krisko, M.; O’Connor, T.; Serianni, A. S.
J. Am. Chem. Soc. 1987, 109, 4464–4472.
18. Sperber, N.; Zaugg, H. E.; Sandstrom, W. M. J. Am.
Chem. Soc. 1947, 69, 915–920.
5-Deoxy- -lyxonohydroxamic acid (10).—
L
The title compound was obtained as a color-
less syrup, yield: 42 mg (100%). IR (KBr) w
1
1669 (CO); 3379 (br NH, OH) cm−1. H
19. Serianni, A. S.; Nunez, H. A.; Barker, R. J. Org. Chem.
1980, 45, 3329–3341.
NMR (D2O, 200 MHz): l 4.15 (d, J2,3 6 Hz, 1
H, H-2); 3.98 (qd, J3,4 3 Hz, 1 H, H-4); 3.59
(dd, 1 H, H-3); 1.19 (d, J4,5 7 Hz, 1 H, H-5).
13C NMR (D2O, 62.5 MHz): l 171.4 (C-1);
75.6, 72.1, 67.2 (C-2, C-3, C-4); 19.5 (C-5).
MS (negative mode): 164.0 [M−H]−. Anal.
Calcd for C5H11NO5: C, 36.36; H, 6.71;
Found: C, 35.61; H, 6.87.
20. Hardegger, E.; Kreis, K.; El Khadem, H. Hel6. Chim.
Acta 1951, 34, 2343–2348.
21. Ga¨tzi, K.; Reichstein, T. Hel6. Chim. Acta 1937, 20,
1298–1303.
22. Moore, S.; Link, K. P. J. Biol. Chem. 1940, 133, 292–
311.
23. Humphlett, W. J. Carbohydr. Res. 1967, 4, 157–164.
24. Clark, E. P. J. Biol. Chem. 1922, 54, 65–73.
25. Kaiser, H.; Keller-Schierlein, W. Hel6. Chim. Acta 1981,
64, 4464–4472.

204
L. Salmon et al. / Carbohydrate Research 335 (2001) 195–204
26. Chirgwin, J. M.; Noltmann, E. A. J. Biol. Chem. 1975,
250, 7272–7276.
27. Larsen, I. K. Acta Crystallogr., Sect. B 1988, 44, 527–
533.
28. Bracher, B. H.; Small, R. W. H. Acta Crystallogr., Sect.
B 1970, 26, 1705–1709.
29. Schouten, A.; Kanters, J. A.; Kroon, J.; Comini, S.;
Looten, P.; Mathlouthi, M. Carbohydr. Res. 1998, 312,
131–137.
30. Andre´, C.; Luger, P. Acta Crystallogr., Sect. B 1993, 49,
375–382.
31. Larsen, I. K.; Trickey, P. Acta Crystallogr., Sect. C 1995,
51, 125–127.
32. El Yazal, J.; Pang, Y.-P. J. Phys. Chem. A 1999, 103,
8346–8350.
33. Nin˜o, A.; Mun˜oz-Caro, C.; Senent, M. L. J. Mol. Struct.
(Theochem.) 2000, 530, 291–300.
34. Remko, M.; Sefc´ıkova´, J. J. Mol. Struct. (Theochem.)
2000, 528, 287–296.
35. Ojala, C. R.; Ostman, J. M.; Ojala, W. H.; Hanson, S. E.
Carbohydr. Res. 2001, 331, 319–325.
36. Nogushi, K. K. O.; Ohno, S.; Hidano, T.; Wakiuchi, N.;
Tarui, T.; Tamaki, H.; Kishihara, S.; Fujii, S. Carbohydr.
Res. 2000, 328, 241–248.
37. Ojala, W. H.; Ostman, J. M.; Ojala, C. R. Carbohydr.
Res. 2000, 326, 104–112.
38. Ko¨ll, P.; Bischoff, M.; Morf, M.; Zimmer, B.; Kopf, J.
Carbohydr. Res. 1993, 248, 45–53.
39. Zinbo, M.; Sherman, W. R. J. Am. Chem. Soc. 1970, 92,
2105–2114.
40. Burla, M. C.; Camalli, M.; Cascarano, G.; Giacovazzo,
C.; Polidori, G.; Spagna, R.; Viterbo, D. J. Appl. Cryst.
1989, 22, 389–393.
41. Watkin, D. J.; Carruthers, J. R.; Betteridge, P. W. CRYS-
TALS—User Guide; Chemical Crystallography Labora-
tory, University of Oxford: Oxford, UK, 1985.
42. Walker, N.; Stuart, D. Acta Crystallogr., Sect. A 1983,
39, 158–166.
43. Prince, E. Mathematical Techniques in Crystallography;
Springer–Verlag: Berlin, 1982.
44. Pearce, L. J.; Watkin, D. J.; Prout, C. K. CAMERON. A
Program for Plotting Molecular Structures; Chemical
Crystallography Laboratory, University of Oxford: Ox-
ford, UK, 1992.
45. Cromer, D. T.; Waber, J. T. International Tables for
X-ray Crystallography; Kynoch Press: Birmingham, UK,
1974; Vol. IV, pp. 99–101 Table 2.2B%.