Application of appel reaction to the primary alcohol groups of fructooligosaccharides: Synthesis of 6,6′,6′′-trihalogenated 1-kestose derivatives

Article abstract of DOI:10.24820/ark.5550190.p010.706

1-kestose (O-β-D-fructofuranosyl-(2→1)-β-D-fructofuranosyl-(2→1)-α-D-glucopyranoside) is a potential short chain fructooligosaccharide with an inulin-type skeleton. Halogenation of 1-kestose was conducted via the Appel reaction with the use of carbon tetrahalide (CBr₄ or CCl₄) and triphenylphosphine, which was then followed by conventional acetylation. The per-O-acetylated form of 6,6’,6’’-trihalogenated derivatives of 1-kestose were conveniently isolated. Further deprotection of the per-O-acetylated form resulted in 6-, 6’-, and 6’’-trihalogenated derivatives. The structure elucidation by one- and two-dimensional nuclear magnetic resonance established that halogenations are specific at the 6-, 6’-, and 6’’-position of 1-kestose primary alcohols.

Full text of DOI:10.24820/ark.5550190.p010.706

The Free Internet Journal
for Organic Chemistry
Paper
Archive for
Arkivoc 2018, part vii, 0-0
Organic Chemistry
to be inserted by editorial office
Application of Appel reaction to the primary alcohol groups of
fructooligosaccharides: Synthesis of 6,6′,6′′-trihalogenated 1-kestose derivatives
Zetryana Puteri Tachrim,a Tadashi Nakamura,b Yasuko Sakihama,a Yasuyuki Hashidoko,a and Makoto
Hashimoto*a
aDivision of Applied Science, Graduate School of Agriculture, Hokkaido University, Sapporo, Japan
bDepartment of Agricultural and Life Science, Obihiro University of Agriculture and Veterinary Medicine,
Obihiro, Japan
Email: hasimoto@abs.agr.hokudai.ac.jp
Received 07-30-2018
Accepted 10-19-2018
Published on line 11-18-2018
Abstract
1-kestose (O-β-D-fructofuranosyl-(2→1)-β-D-fructofuranosyl-(2→1)-α-D-glucopyranoside) is a potential short
chain fructooligosaccharide with an inulin-type skeleton. Halogenation of 1-kestose was conducted via the
Appel reaction with the use of carbon tetrahalide (CBr4 or CCl4) and triphenylphosphine, which was then
followed by conventional acetylation. The per-O-acetylated form of 6,6’,6’’-trihalogenated derivatives of 1-
kestose were conveniently isolated. Further deprotection of the per-O-acetylated form resulted in 6-, 6’-, and
6’’-trihalogenated derivatives. The structure elucidation by one- and two-dimensional nuclear magnetic
resonance established that halogenations are specific at the 6-, 6’-, and 6’’-position of 1-kestose primary
alcohols.
Keywords: Appel reaction; nuclear magnetic resonance (NMR); primary alcohol; regioselective halogenations;
1-kestose; fructooligosaccharides
DOI: https://doi.org/10.24820/ark.5550190.p010.706
Page 1
©ARKAT USA, Inc

Arkivoc 2018, vii, 0-0
Tachrim, Z. P. et al.
Introduction
Inulin-type short-chain fructooligosaccharides (FOS) are fructose oligomers that consist of a terminal glucosyl
unit and two to five fructosyl units. They are recognized as prebiotic indigestible organosaccharides1 and due
to this property, FOS demand has been increasing in the food industry.2 1-Kestose (1, Figure 1) is one of FOS
that has a β-D-fructofuranosyl group on O-1 of the D-fructosyl moiety of sucrose (2, Figure 1). 1-Kestose 1 is
naturally found in honey and some plants belonging to the Amaryllidaceae family.3–5 The interest in 1-kestose
1, as a low-calorie food ingredient, continues to increase due to its sweetening power. FOS syrup enriched by
1-kestose 1 can be used as alternative sweetener for diabetics.6 To produce 1-kestose 1, the enzyme derived
from the leaves of sugar beets have demonstrated transfructosylation activity in the present of sucrose 2;
thus, the products of transfer were mainly 1-kestose 1 with smaller proportions of other oligosaccharides.7
Commercial cellulolytic enzymes have also been studied for preparation of FOS with high 1-kestose
concentrations.2
Figure 1. Structure of 1-kestose (1), sucrose (2), and undecaacetate of 1-kestose (3).
Since the specific substitution of sucrose primary hydroxyl groups by chloride enhances the sweetness
activity,8 halogenated carbohydrate derivatives can potentially be used as an alternative sweetener. The direct
halogenation of the hydroxyl groups of carbohydrate is a convenient method to achieve the synthesis of
halogenated carbohydrate on its primary hydroxyl groups. Thus, selective halogenation for carbohydrate has
become an area of interest in organic chemistry. The Appel reaction is one of the most important reactions
used to convert the primary hydroxyl group into a halo methylene group in the presence of
triphenylphosphine and carbon tetrahalide.9 This method was used for selective halogenation of primary
hydroxyl groups over secondary hydroxyl groups in carbohydrates. Halogenation (particularly for chlorination
and bromination) of primary alcohols of sucrose 210—the main precursor for enzyme-catalyst generation of 1-
kestose 1—has been studied since the 1970s11, but the reactivity of primary hydroxyl groups attained using
the Appel reaction has not been completely identified due to limitation of structure analysis. Recently, we
reported that sucrose 2 can be selectively brominated and chlorinated using the Appel reaction at only the 6-
and 6’- position with no halogenation at 1’-position supported by one- and two-dimensional (1D and 2D,
respectively) NMR analysis.12 As for trisaccharide modification, halogenation of the primary hydroxyl groups of
raffinose (O-α-D-galactopyranosyl-(1→6)-α-D-glucopyranosyl-(1→2)-β-D-fructofuranoside) has been reported
previously.13 Raffinose reacted with sulfuryl chloride and the authors found that the chlorinated proportion at
the primary position was followed by chlorination at the 4-position of the galactopyranosyl moiety secondary
alcohols. To date, no study has been attempted to substitute primary alcohols of FOS using halogens. In midst
of FOS, 1-kestose 1 has shown relatively high sweetness activity and is already commercially available.
Therefore, its primary alcohol modification would potentially increase the synthesis of alternative sweeteners.
Page 2
©ARKAT USA, Inc

Arkivoc 2018, vii, 0-0
Tachrim, Z. P. et al.
In this study, we aimed to synthesize halogenated 1-kestose 1 at the primary position by using the Appel
reaction to comprehensively study the structure elucidation supported by 1D and 2D NMR analysis.
Results and Discussion
Direct substitution of 1-kestose 1 with 4.7 equiv. carbon tetrahalide (bromide or chloride) and 9.1 equiv.
o
triphenylphosphine at 70 C for two hours (Scheme 1) produced a complex mixture with complicated 1H-NMR
analysis due to observation of overlap signals, especially the modified primary centers. The mixture was
separated after conventional acetylation. Further purification was conducted using an ether and hexane
system as the representative mobile phase to isolate halogenated carbohydrate. The ethyl acetate or
dichloromethane system cannot be used for the halogenated proportion.12 The mixture produced halogenated
1-kestose derivatives in the pre-O-acetylated form (bromination 4 and chlorination 5, Scheme 1).
Scheme 1. Halogenation of 1-kestose (1) via Appel reaction with Ph3P and carbon tetrahalide to produce pre-
O-acetylated form of halogenated 1-kestose derivatives (bromination 4 and chlorination 5).
1
The H NMR analysis for peracetylated 4 and 5 showed two regions that contribute to the proton
resonance of aliphatic sugar groups in the downfield region and halogenated methylene groups located in the
upfield region. Based on 13C NMR, three halogenated methylene in the upfield region were easily determined
in the halogenated proportion in the 1-kestose derivatives, but the halogenated position remained unclear.
Therefore, 2D NMR (COSY, HETCOR, HMQC, HMBC, NOESY, and TOCSY) was used to determine the
halogenated position. For this purpose, CDCl312,14 was used as the solvent to increase the visibility of the spin
system during the analysis. Undecaacetate 1-kestose3,15 (3, Figure 1, Figures SM-17–24 for detail NMR analysis
and Table SM-1 for 1D and 2D NMR and comparison of structure elucidation with the literature) was identified
and used to understand the sugar skeleton of halogenated 1-kestose derivatives.
1
During brominated position investigation of the 1-kestose derivative, H-NMR of compound 4 showed
eight proton signals in the upfield region. The excess of two protons corresponds to the H-1’ glycosidic bond of
O-β-D-fructofuranosyl-(2→1)-β-D-fructofuranosyl, which showed considerable geminal coupling with a typical
pair of doublets at δ = 3.79 and 3.73 ppm (J = 10.3 Hz). The correlation between H-1’a and H-1’b with C-2’ in
HMBC, supported by HETCOR and HMQC, distinguished this spin system (Figures SM-4–6). The same tendency
was also demonstrated by the undecaacetate of 1-kestose 3, which supported the glycosidic bond signals. The
additional six proton signals in the upfield region highlighted the specific brominated proportion of compound
4. A clear pair of doublets at δ = 3.54 and 3.40 ppm (J = 2.3, 11.5 Hz and J = 6.3, 11.5 Hz, respectively) were
1
observed by H-NMR. The geminal coupling for these spins systems indicated the brominated methylene of
the glucose ring at the 6-position, which was easily distinguished by its correlation with H-5 in COSY and with
Page 3
©ARKAT USA, Inc

Arkivoc 2018, vii, 0-0
Tachrim, Z. P. et al.
H-4 and H-5 in TOCSY (Figures SM-3 and SM-8). Moreover, the carbon brominated terminal at the 6-position
(δ = 31.1 ppm) was also identified, as supported by HMBC analyses. The observation of the long range
correlation of C-6 with H-4 clearly distinguished brominated position which supported by HMQC and HETCOR.
The glucose ring’s protons and carbons signals were distinguished from fructose rings signals, especially at the
C-3, C-4, and C-5 positions, using TOCSY and NOESY (Figures SM-7 and SM-8), which was also, supported by
HETCOR, HMQC, and HMBC analyses.
Among 1-kestose’s fructose rings, there are three possible primary alcohols at the 6’-, 1’’, and 6’’-
position that are readily brominated. The upfield region of 1H-NMR typically showed two types of doublet at δ
= 3.66 ppm (J = 6.9 Hz) and 3.69 ppm (J = 7.4 Hz), indicating bromination occurred only at the 6’- and 6’’-
position of compound 4, respectively. The correlation between H-6’ with H-5’ in COSY and H-6′ with H-4’ and
H-5’ in TOCSY differentiated the bromination at the 6’-position (Figures SM-3 and SM-8). Similarly, brominated
6’’-position of compound 4 showed these particular correlations. The brominated terminal of C-6’ at δ = 32.0
ppm and C-6’’at δ = 32.7 ppm was determined mostly using HMBC with the observation of a cross peak of
these carbons at H-4’ and H-4’’, respectively (Figures SM-6). HETCOR and HMQC coupling between the protons
and carbon at the 6’- and/or 6’’-position support the identification of bromination position.
The aliphatic sugar groups between the two fructose rings of compound 4 were differentiated using
TOCSY and NOESY, which was supported by COSY, HETCOR, HMQC, and HMBC analyses (Figures SM-3–8).
1
Based on H- and 13C-NMR, supported by 2D NMR analysis, the 1’’-position of compound 4 showed no
bromination. The 1’’-position of the fructose ring is known to be a neopentyl-like10,12 terminal having the least
reactive proportion. This phenomena also occurred when sucrose was used.12,16,17 According to these
approaches, the bromination product of 1-kestose was determined to be 1’’,2,3,3’,3’’,4,4’,4’’-octa-O-acetyl-
6,6’,6’’-tribromo-6,6’,6’’-trideoxy-1-kestose (4).
1
For the chlorination product of 5, the spin system in H-NMR showed eight proton signals in the upfield
region. Unlike the previous bromination product 4 or undecaacetate 1-kestose 3, the H-1’ of compound 5
exhibited no geminal coupling and presented as a singlet at 3.75 ppm supported by HMBC, HETCOR, and
HMQC, (Figures SM-12–14). The chlorinated proportion at the 6-position was shown as a pair of double
doublets at δ = 3.68 and 3.57 ppm (J = 2.3, 12.0 Hz and J = 5.7, 12.0 Hz, respectively). A clear correlation of this
chlorinated methylene of the glucose ring with H-5 in COSY and H-4 and H-5 in TOCSY was observed (Figures
SM-11 and SM-16). As for the 1H-NMR spin system of 6’- and 6’’-chloro of compound 5, the overlap signals at δ
= 3.82–3.78 ppm demonstrated its correlation with H-5’ and H-5’’ in COSY and H-4’, H-5’, H-4’’, and H-5’’ in
TOCSY. The brominated terminal of C-6’ at δ = 44.4 ppm and C-6’’ at δ = 44.6 ppm highlighted the correlated
cross peak of these carbons to H-4’ and H-4’’, respectively. HMBC strongly distinguished the chlorinated 6’-
and 6′′-position of compound 5 (Figures SM-14). These results support that the chlorination product of 1-
kestose is 1′′,2,3,3′,3′′,4,4′,4′′-octa-O-acetyl-6,6′,6′-trichloro-6,6′,6′′-trideoxy-1-kestose (5). As with brominated
compound 4, chlorinated compound 5 showed no substitution at the 1′′-position. The presence of Br and Cl
isotopic peak in ESI-MS of compounds 4 and 5 indicated tri-halogenated proportion of 1-kestose, respectively,
which supported our analysis.
To synthesize and elucidate the novel 6,6′,6′′-trihalogenated 1-kestose derivatives, deacetylation was
then conducted. Since suitable deacetylation conditions are generated using saturated ammonium in
methanol for sucrose,12 the undecaacetate of 1-kestose (3) was first tested for deacetylation and produced 1-
kestose (1) within a satisfactory yield (Figures SM-41–48 for NMR analysis). Deacetylation using other
methods, such as sodium methoxide, cannot be used for 1-kestose halogenated derivatives. The per-O-
acetylated trihalogenated derivatives of 4 and 5 were underwent deacetylation, which resulted in compound 6
and 7 (Scheme 2). It was complicated to distinguish the halogenated proportion from the deacetylated
Page 4
©ARKAT USA, Inc

Arkivoc 2018, vii, 0-0
Tachrim, Z. P. et al.
compound 6 and/or 7, since the halogenated methylene spin system at H-6, H-6′, and H-6′′ overlapped with
1
other signals such as H-3, H-1′, and/or H-1′′ in H-NMR. Hence, the HMBC, HMQC, and HETCOR analyses,
supported by COSY, TOCSY, and NOESY, are needed to elucidate the complete structure of deacetylated
compounds 6 and 7 (Figures SM-27–32 and SM-35–40). However, all the deacetylated trihalogenated 1-
kestose derivatives (6 and 7) structures and halogenation sites were consistent with those in the
corresponding per-O-acetylated forms (4 and 5).
Scheme 2. Deacetylation of per-O-acetylated 1-kestose derivatives (3–5) to result in 1-kestose (1) and
deacetylated trihalogenated 1-kestose derivatives (6 and 7)
Conclusions
We studied the halogenation of 1-kestose using Appel reactions to synthesize trihalogenated 1-kestose at the 6-,
6′-, and 6′′-position. Isolation and structure elucidation were easily completed using the per-O-acetylated form
which the 1D and 2D NMR supported the halogenation position. The synthesis and structure elucidation of
novel compounds 4 and 5 contribute to the introduction of primary hydroxyl groups into FOS which can
potentially be used as low-calorie sweeteners in the future.
Experimental Section
General. All reagents used were of analytical grade. NMR spectra were obtained in CDCl3 or D2O by JEOL
ECA500 (500 and 125 MHz) spectrometer (JEOL, Tokyo, Japan). Optical rotations were measured at 23 oC on a
JASCO DIP370 polarimeter (JASCO, Tokyo, Japan). HRMS spectra were obtained with a Waters UPLC ESI-TOF
mass spectrometer (Waters, Milford, CT, USA).
General procedure for Appel reaction: bromination. Solution of 1-kestose (1, 0.10 g, 0.198 mmol) in pyridine
(2 mL) was cooled in an ice bath and treated with triphenylphosphine (0.47 g, 1.8 mmol, 9.1 equiv), followed
by dropwise addition of a solution of carbon tetrabromide (0.30 g, 0.93 mmol, 4.7 equiv) in pyridine (0.93 mL).
The reaction mixture was stirred at 70 °C for 2 h. After cooling the mixture in an ice bath, acetic anhydride (3
mL, 32.0 mmol) and pyridine (2 mL) were added, and the mixture was stirred overnight at room temperature.
The solvent was evaporated and the residue was partitioned between CH2Cl2 and water. The organic layer was
washed with 1 M HCl and brine, dried over MgSO4, reduced the solvent under high pressure and then purified
by column chromatography on silica gel by elution with hexane/diethyl ether (1:6 and 1:10) to yield
Page 5
©ARKAT USA, Inc

Arkivoc 2018, vii, 0-0
Tachrim, Z. P. et al.
1′′,2,3,3′,3′′,4,4′,4′′-octa-O-acetyl-6,6′,6′′-tribromo-6,6′,6′′-trideoxy 1-kestose (4; 0.0516 g, 25%) as colorless
1
amorphous mass. [α]D +22.4 (c = 1.0, CHCl3). H-NMR (500 MHz, CDCl3) δ: 5.70 (1H, d, J3′,4′ 8.0 Hz, H-3′), 5.67
(1H, d, J1,2 = 4.0 Hz, H-1), 5.50 (1H, d, J3′′,4′′ 6.3 Hz, H-3′′), 5.48–5.42 (2H, m, H-3, H-4′), 5.32 (1H, t, J3′′,4′′ 6.3 Hz,
H-4′′), 5.03 (1H, t, J3,4 9.7 Hz, H-4), 4.95 (1H, dd, J1,2 4.0 Hz, J2,3 10.3 Hz, H-2), 4.35–4.28 (2H, m, H-5, H-5′), 4.24
(2H, s, H-1′′), 4.22–4.19 (1H, m, H-5′′), 3.79 (1H, d, J1′a,1′b 10.3 Hz, H-1′a), 3.73 (1H, d, J1′a,1′b 10.3 Hz, H-1′b), 3.69
(2H, d, J5′′,6′′ 7.4 Hz, H-6′′), 3.66 (2H, d, J5′,6′ 6.9 Hz, H-6′), 3.54 (1H, dd, J5,6a 2.3 Hz, J6a,6b 11.5 Hz, H-6a), 3.40 (1H,
dd, J5,6b 6.3 Hz, J6a,6b 11.5 Hz, H-6b), 2.16–2.15 (6H, m, 2 x CH3), 2.14 (3H, s, CH3), 2.11 (3H, s, CH3), 2.09–2.07
(9H, m, 3 x CH3), 2.02 (3H, s, CH3) ppm. 13C-NMR (125 MHz, CDCl3) δ: 170.2, 170.0, 170.0, 169.8, 169.5 (C=O),
103.8 (C-2′), 103.1 (C-2′′), 90.1 (C-1), 80.7 (C-5′′), 79.8 (C-5′), 78.1 (C-4′′), 77.2 (C-3′′), 76.4 (C-4′), 75.8 (C-3′),
70.8 (C-4), 69.7 (C-2), 69.3 (C-3 & C-5), 62.0 (C-1′′), 61.5 (C-1′), 32.7 (C-6′′), 32.0 (C-6′), 31.1 (C-6), 20.8, 20.8,
20.7, 20.8, 20.6, 20.5, 20.5, 20.5 (CH3) ppm. HRMS (ESI): calcd. for C34H45O21Br3Na 1050.9881, 1052.9860 [M +
Na]+; found 1050.9899, 1052.9880.
Chlorination. Chlorination was similar to bromination as described above, except carbon tetrabromide in
pyridine was replaced with carbon tertrachloride (0.09 mL, 0.93 mmol, 4.7 equiv.) to yield 1′′,2,3,3′,3′′,4,4′,4′′-
octa-O-acetyl-6,6′,6′′-trichloro-6,6′,6′′-trideoxy-1-kestose (5; 0.0514 g, 28%) as colorless amorphous mass.
[α]D +26.6 (c = 1.0, CHCl3). 1H-NMR (500 MHz, CDCl3) δ: 5.70–5.66 (2H, m, H-1, H-3′), 5.50 (1H, d, J3′′,4′′ 6.3 Hz,
H-3′′), 5.48–5.41 (2H, m, H-3, H-4′), 5.33 (1H, t, J3′′,4′′ = 6.3 Hz, H-4′′), 5.06 (1H, t, J3,4 = 9.7 Hz, H-4), 4.93 (1H, dd,
J1,2 3.7 Hz, J2,3 10.6 Hz, H-2), 4.38–4.32 (1H, m, H-5), 4.28–4.22 (3H, m, H-1′′, H-5′), 4.17 (1H, q, J5′′,6′′ 6.5 Hz, H-
5′′), 3.82–3.78 (4H, m, H-6′, H-6′′), 3.75 (2H, s, H-1′), 3.68 (1H, dd, J5,6a 2.3 Hz, J6a,6b 12.0 Hz, H-6a), 3.57 (1H, dd,
J5,6b 5.7 Hz, J6a,6b 12.0 Hz, H-6b), 2.16 (3H, s, CH3), 2.15–2.13 (6H, m, 2 x CH3), 2.11 (3H, s, CH3), 2.09 (3H, s CH3),
2.08 (3H, s, CH3), 2.07 (3H, s, CH3), 2.02 (3H, s, CH3). 13C-NMR (125 MHz, CDCl3) δ: 170.2, 170.1, 170.0, 170.0,
169.7, 169.5 (C=O), 103.8 (C-2′), 103.1 (C-2′′), 90.0 (C-1), 80.6 (C-5′′), 79.9 (C-5′), 77.2 (C-4′′), 76.9 (C-3′′), 75.7
C-4′), 75.5 (C-3′), 69.8 (C-5), 69.7 (C-2 & C-4), 69.4 (C-3), 62.2 (C-1′′), 61.6(C-1′), 44.6 (C-6′′), 44.4 (C-6′), 43.2 (C-
6), 20.8, 20.7, 20.7, 20.6, 20.5, 20.5 (CH3) ppm. HRMS (ESI): calcd. for C34H45O21Cl3Na 919.1387 [M + Na]+;
found 919.1395.
1′′,2,3,3′,3′′,4,4′,4′′,6,6′,6′′-Undeca-O-acetyl-deoxy-1-kestose (3).15,18 Solution of 1-kestose (1, 0.03 g, 0.06
mmol) in pyridine (15 mL) was treated with acetic anhydride (1 mL, 10.7 mmol), and the mixture was stirred
overnight at room temperature. The solvent was removed under reduced pressure and subjected to column
chromatography on silica gel by elution with chloroform to yield 3 (0.055 g, 96%) as a colorless liquid. [α]D +
30.2 (c = 1.0, CHCl3); Ref. 16 [α]D +31.8 (c = 3.7, CHCl3). 1H-NMR (500 MHz, CDCl3) δ: 5.75 (1H, d, J1,2 3.4 Hz, H-1),
5.69 (1H, d, J3′,4′ 8.0 Hz, H-3′), 5.48 (1H, d, J3′′,4′′ 6.9 Hz, H-3′′), 5.46 (1H, t, J3′,4′ 8.0 Hz, H-4′), 5.42 (1H, t, J3,4 9.7
Hz, H-3), 5.34 (1H, t, J3′′,4′′ 6.9 Hz, H-4′′), 5.08 (1H, t, J3,4 9.7 Hz, H-4), 4.91 (1H, dd, J1,2 3.4 Hz, J2,3 10.3 Hz, H-2),
4.39–4.33 (3H, m, H-5, H-6′′), 4.33–4.24 (3H, m, H-6a, H-6′), 4.24–4.20 (3H, m, H-1′′, H-5′), 4.20–4.14 (2H, m, H-
6b, H-5′′), 3.69 (1H, d, J1′a,1′b 9.2 Hz, H-1′a), 3.63 (1H, d, J1′a,1′b 9.2 Hz, H-1′b), 2.19–2.14 (3H, m, CH3), 2.13–2.12
(6H, m, 2 x CH3), 2.11–2.09 (15H, m, 5 x CH3), 2.06 (3H, s, CH3), 2.04 (3H, s, CH3), 2.01 (3H, s, CH3) ppm. 13C-
NMR (125 MHz, CDCl3) δ: 170.7, 170.6, 170.5, 170.1, 169.9, 169.7, 169.6 (C=O), 103.4 (C-2′), 102.9 (C-2′′), 89.2
(C-1), 78.4 (C-5′′), 77.8 (C-5′), 76.5 (C-3′′), 75.5 (C-4′′), 74.9 (C-3′), 73.7 (C-4′), 70.0 (C-2), 69.8 (C-3), 68.2 (C-5),
68.2 (C-4), 63.7 (C-6′′), 63.2 (C-6′), 62.7 (C-1′′), 62.2 (C-1′), 61.7(C-6), 20.8, 20.7, 20.7, 20.6, 20.6, 20.5 (CH3)
ppm. HRMS (ESI): calcd. for C40H54O27Na 989.2750 [M + Na]+; found 989.2797.
General procedure for deacetylation of per-O-acetylated halodeoxy-1-kestose derivatives. Solutions of per-
O-acetylated halodeoxy-1-kestose derivatives (3–5, 0.05 g) in 3 mL dry methanol were treated with 1.5 mL
saturated ammonia in methanol and stirred overnight at room temperature. The solvent was removed under
reduced pressure and subjected to column chromatography on silica gel (pre-washed by methanol) by elution
Page 6
©ARKAT USA, Inc

Arkivoc 2018, vii, 0-0
Tachrim, Z. P. et al.
with methanol/chloroform (3:1). The method produced the corresponding halodeoxy-1-kestose derivatives 1,
6, and 7.
1
1-Kestose3 (1, 0.034 g, 97%). H-NMR (500 MHz, D2O) δ: 5.37 (1H, d, J1,2 4.0 Hz, H-1), 4.22 (1H, d, J3′,4′ 8.6 Hz,
H-3′), 4.13 (1H, d, J3′′,4′′ 8.6 Hz, H-3′′), 4.02 (1H, t, J3′′,4′′ 8.6 Hz, H-4′′), 3.98 (1H, t, J3′,4′ 8.6 Hz, H-4′), 3.83–3.80
(2H, m, H-5′, H-5′′), 3.79–3.71 (8H, m, H-1′a, H-5, H-6, H-6′, H-6′′), 3.70–3.60 (4H, m, H-3, H-1′b, H-1′′), 3.48 (1H,
dd, J1,2 4.0 Hz, J2,3 9.7, H-2), 3.41 (1H, t, J3,4 9.7 Hz, H-4) ppm. 13C-NMR (125 MHz, D2O) δ: 104.2 (C-2′′), 103.7 (C-
2′), 92.9 (C-1), 81.6 (C-5′), 81.5 (C-5′′), 77.0 (C-3′ & C-3′′), 74.9 (C-4′′), 74.2 (C-4′), 73.0 (C-3), 72.8 (C-5), 71.6 (C-
2), 69.6 (C-4), 62.7 (C-6′′), 62.6 (C-6′), 61.3 (C-1′), 60.8 (C-1′′), 60.5 (C-6) ppm.
1
6,6′,6′′-Tribromo-6,6′,6′′-trideoxy-1-kestose (6, 0.030 g, 81%). [α]D = + 24.8 (c 1.0, MeOH). H-NMR (500 MHz,
D2O) δ: 5.36 (1H, d, J1,2 4.0 Hz, H-1), 4.27 (1H, d, J3′,4′ 8.6 Hz, H-3′), 4.16 (1H, d, J3′′,4′′ 8.6 Hz, H-3′′), 4.09–4.05
(2H, m, H-4′, H-4′′), 4.04–3.98 (3H, m, H-5, H-5′, H-5′′), 3.87 (1H, d, J1′a,1′b 10.3 Hz, H-1′a), 3.79–3.72 (3H, m, H-
6a, H-6′′), 3.72–3.65 (5H, m, H-1′b, H-1′′, H-6b, H-3), 3.64–3.59 (2H, m, H-6′), 3.52 (1H, dd, J1,2 4.0 Hz, J2,3 10.0,
Hz, H-2), 3.43 (1H, t, J3,4 10.0 Hz, H-4) ppm. 13C-NMR (125 MHz, D2O) δ: 103.9 (C-2′′), 103.3 (C-2′), 92.9 (C-1),
80.6 (C-5′), 80.4 (C-5′′), 77.4 (C-4′′), 76.9 (C-3′′), 76.4 (C-3′), 76.3(C-4′), 72.1(C-3), 71.4 (C-4), 71.1(C-5), 71.0 (C-
2), 59.9 (C-1′, C-1′′), 34.2 (C-6), 33.5 (C-6′), 33.5 (C-6′′) ppm. HRMS (ESI): calcd. for C18H29Br3O13Na 714.9035,
716.9015 [M + Na]+; found 714.9053, 716.9034.
1
6,6′,6′′-Trichloro-6,6′,6′′-trideoxy-1-kestose (7, 0.032 g, 85%). [α]D = +26.6 (c 1.0, MeOH). H-NMR (500 MHz,
D2O) δ: 5.36 (1H, d, J1,2 4.0 Hz, H-1), 4.26 (1H, d, J3′,4′ 8.6 Hz, H-3′), 4.16 (1H, d, J3′′,4′′ 8.6 Hz, H-3′′), 4.11–4.05
(3H, m, H-4′, H-4′′, H-5), 4.01–3.94 (2H, m, H-5′, H-5′′), 3.89–3.73 (7H, m, H-6, H-6′, H-6′′, H-1′a), 3.71–3.60 (4H,
13
m, H-1′b, H-1′′, H-3), 3.52 (1H, dd, J1,2 4.0 Hz, J2,3 9.7 Hz, H-2), 3.47 (1H, t, J3,4 9.7 Hz, H-4) ppm. C-NMR (125
MHz, D2O) δ: 103.9 (C-2′′), 103.3 (C-2′), 92.8 (C-1), 80.6 (C-5′), 80.5 (C-5′′), 76.6 (C-3′′), 76.4 (C-3′), 76.2 (C-4′′),
75.3 (C-4′), 72.2 (C-3), 71.6 (C-4), 71.0 (C-5), 70.1 (C-2), 60.2 (C-1′), 60.0 (C-1′′), 45.4 (C-6′′), 45.2 (C-6′), 44.3 (C-
6) ppm. HRMS (ESI): calcd. for C18H29Cl3O13Na 581.0571, 583.0542 [M + Na]+; found 581.0587, 583.0568.
Acknowledgements
Zetryana Puteri Tachrim thanks LPDP (Indonesia Endowment Fund for Education) for financial support. This
research was partially supported by the Ministry of Education, Science, Sports, and Culture Grant-in-Aid for
Scientific Research (C), (17K0194007 to Makoto Hashimoto). Part of this work was performed under the
Cooperative Research Program of “Network Joint Research Center for Materials and Devices”.
Supplementary Material
One- and two-dimensional nuclear magnetic resonance (1D and 2D NMR, respectively) figures for all synthetic
compounds are available online.
Page 7
©ARKAT USA, Inc

Arkivoc 2018, vii, 0-0
Tachrim, Z. P. et al.
References
1. Ballesteros, A.; Plou, F. J.; Alcalde, M.; Ferrer, M.; Garcıa-Arellano, H.; Reyes-Duarte, D.; Ghazi, I. In
Biocatalysis in the pharmaceutical and biotechnology industries; R.N. Patel, Ed.; CRC Press: Boca Raton,
2007; pp 463–488.
2. Vega, R.; Zúniga-Hansen, M. E. Bioresour. Technol., 2011, 102, 10180.
https://doi.org/10.1016/j.biortech.2011.09.025
3. Calub, T. M.; Waterhouse, A. L.; Chatterton, N. J. Carbohydr. Res., 1990, 199, 11.
https://doi.org/10.1016/0008-6215(90)84088-C
4. Siddiqui, I. R.; Purgala, B. J. Apic. Res., 1968, 7, 51.
https://doi.org/10.1080/00218839.1968.11100188
5. Hammer, H. Acta Chem. Scand., 1968, 22, 197.
https://doi.org/10.3891/acta.chem.scand.22-0197
6. Yun, J.W. Enzyme Microb. Technol., 1996, 19, 107.
https://doi.org/10.1016/0141-0229(95)00188-3
7. Allen, P. J.; Bacon, J. S. D. Biochem. J., 1956, 63, 200.
https://doi.org/10.1042/bj0630200
8. Hough, L.; Khan, R. Trends Biochem. Sci., 1978, 3, 61.
https://doi.org/10.1016/S0968-0004(78)93998-1
9. Appel, R. Angew. Chem. Int. Ed., 1975, 14, 801.
https://doi.org/10.1002/anie.197508011
10. Jarosz, S.; Mach, M. Eur. J. Org. Chem., 2002, 769.
https://doi.org/10.1002/1099-0690(200203)2002:5<769::AID-EJOC769>3.0.CO;2-F
11. Kashem, A.; Anisuzzaman, M.; Whistler, R. L. Carbohydr. Res., 1978, 61, 511.
https://doi.org/10.1016/S0008-6215(00)86063-2
12. Tachrim, Z. P.; Wang, L.; Yoshida, T.; Muto, M.; Nakamura, T.; Masuda, K.; Hashidoko, Y.; Hashimoto, M.
ChemistrySelect, 2016, 1, 58.
https://doi.org/10.1002/slct.201500003
13. Hough, L.; Richardson, A. C.; Salam, M. A. Carbohydr. Res., 1979, 71, 85.
https://doi.org/10.1016/S0008-6215(00)86063-2
14. Lees, W. J.; Whitesides, G. M. J. Am. Chem. Soc. 1993, 115, 1860.
https://doi.org/10.1021/ja00058a034
15. Pejin, B.; Iodice, C.; Tommonaro, G.; Sabovljevic, M.; Bianco, A.; Tesevic, V.; Vajs, V. De Rosa, S. Nat. Prod.
Res. 2012, 26, 209.
https://doi.org/10.1080/14786419.2010.535163
16. Wang, L.; Hashidoko, Y.; Hashimoto, M. J. Org. Chem., 2016, 81, 4464
https://doi.org/10.1021/acs.joc.6b00144
17. Wang, L.; Tachrim, Z. P.; Kurokawa, N.; Ohashi, F.; Wakasa, H.; Sakihama, Y.; Hashidoko, Y.; Suzuki, T.;
Hashimoto, M. Arkivoc, 2018, 7, 56.
https://doi.org/10.24820/ark.5550190.p010.668
18. Binkley, W. W.; Horton, D.; Bhacca, N. S. Carbohydr. Res., 1969, 10, 245.
https://doi.org/10.1016/S0008-6215(00)80466-8
Page 8
©ARKAT USA, Inc