2210
G. Lopez et al. / Carbohydrate Research 342 (2007) 2202–2211
(CH2Cl2–MeOH, 20:1) to yield 15 (23 mg, 88%) as a yel-
low solid (CH2Cl2–MeOH, 20:1, Rf = 0.35); mp 123 ꢁC;
(D2O, 500.13 MHz): d 8.17 (d, 2H, J = 9.2 Hz, H-30,
H-50), 7.12 (d, 2H, J = 9.3 Hz, H-20, H-60), 5.78 (s,
1H, H-1a), 5.08 (s, 1H, H-1b), 4.44 (dd, 1H, J1a,2a
<1 Hz, J2a,3a = 2.1 Hz, H-2a), 4.20 (t, 1H,
J4a,5a = 6.5 Hz, H-4a), 4.02 (dd, 1H, J1b,2b <1 Hz,
J2b,3b = 4.4 Hz, H-2b), 3.98 (t, 1H, J4b,5b = 6.3 Hz, H-
4b), 3.81 (dd, 1H, J3a,4a = 5.2 Hz, H-3a), 3.65 (dd, 1H,
J3b,4b = 6.7 Hz, H-3b), 1.29 (d, 3H, H-5a), 1.25 (d, 3H,
20
1
½aꢁD +79.4 (c 1, acetone); H NMR (D2O, 400 MHz): d
8.15 (d, 2H, J = 7.3 Hz, H-30, H-50), 7.11 (d, 2H,
J = 7.4 Hz, H-20, H-40), 6.16 (s, 1H, H-1), 4.21 (ddd,
1H, J2a,3 = 6.3 Hz, J2b,3 = 2.9 Hz, J3,4 = 3.7 Hz, H-3),
3.99–3.95 (m, 1H, H-4), 3.77–3.71 (m, 2H, H-5), 2.24
(dddd, 2H, J1,2a = 1.1 Hz, J1,2b = 4.7 Hz, H-2); 13C
NMR (D2O, 100 MHz): d 161.4 (C-10), 142.9 (C-40),
126.4 (C-30, C-50), 117.6 (C-20, C-60), 102.6 (C-1), 86.2
(C-4), 71.4 (C-3), 63.1 (C-5), 42.5 (C-2). Anal. Calcd
for C11H13NO6: C, 51.77; H, 5.13. Found: C, 51.84;
H, 5.23.
13
H-5b); C NMR (D2O, 125.76): d 161.30 (C-10),
142.20 (C-40), 126.05 (C-30, C-50), 116.68 (C-20, C-60),
106.81 (C-1b), 105.15 (C-1a), 86.93 (C-3a), 81.60 (C-
2b, C-3b), 79.98 (C-2a), 79.27 (C-4a, C-4b), 17.59 (C-
5a, C-5b); HRMS (ESI+) for C16H21NO9Na: m/z
[M+Na]+ calcd 394.1114, found: 394.1117.
4.15. Autocondensation reactions
A 5 mM aqueous solution of 6 or 10 (1.4 mL) was incu-
bated with 12 IU AbfD3 with magnetic stirring for 1 h
at 60 ꢁC. Reactions were quenched by enzyme denatur-
ation at 100 ꢁC for 10 min. After lyophilization, the
residue was dissolved in water (0.1 mL) and the auto-
condensation products were purified by preparative
TLC using 4:1 CH2Cl2–MeOH as the mobile phase.
The desired products were detected by UV absorption
(254 nm), collected from plates and extracted with 1:1
CH2Cl2–MeOH (4 mL). After filtration and freeze-
drying, the isolated disaccharides 16 and 17 were
characterized by NMR and high resolution mass
spectrometry.
Acknowledgements
Funding for G.L. was provided by the French Ministry
of Education, Research and Technology. We wish to
thank J. P. Guegan (ENSCR), S. Sinbandhit (Centre
Regional de Mesures Physiques de l’Ouest, CRMPO,
University of Rennes 1, France), and P. Guenot and
´
´
´
P. Jehan (CRMPO) for recording HRMS spectra.
References
1. Krˇen, V.; Thiem, J. J. Chem. Soc. Rev. 1997, 26, 463–473.
2. Monsan, P.; Paul, F. FEMS Microbiol. Rev. 1995, 16,
187–192.
3. Scigelova, M.; Singh, S.; Crout, D. H. G. J. Mol. Catal. B:
Enzym. 1999, 6, 483–494.
4.16. p-Nitrophenyl 5-deoxy-a-L-arabinofuranosyl-
(1!2)-5-deoxy-a-L-arabinofuranoside (16)
(ꢂ1 mg accumulated from a few preparative plates)
HPTLC (CH2Cl2–MeOH, 4:1) Rf = 0.46; 1H NMR
(D2O, 500.13 MHz): d 8.17 (d, 2H, J = 9.3 Hz, H-30,
H-50), 7.11 (d, 2H, J = 9.3 Hz, H-20, H-60), 5.81 (d,
1H, J1a,2a = 1.7 Hz, H-1a), 5.06 (d, 1H, J1b,2b = 1.2 Hz,
H-1b), 4.24 (dd, 1H, J2a,3a = 4.9 Hz, H-2a), 4.09 (dd,
1H, J4a,5a = 6.4 Hz, J3a,4a = 7.5 Hz, H-4a), 4.03 (dd,
1H, J2b,3b = 3.9 Hz, H-2b), 3.92 (t, 1H, J4b,5b = 6.5 Hz,
H-4b), 3.86 (dd, 1H, H-3a), 3.60 (dd, 1H,
J3b,4b = 6.7 Hz, H-3b), 1.25 (d, 3H, H-5a), 1,07 (d, 3H,
4. Lee, R.; Mansey, D.; Watson, A.; Duncan, K.; Rithner,
C.; McNeil, M. Anal. Biochem. 1996, 242, 1–7.
5. Cote, G. L.; Tao, B. Y. Glycoconjugate J. 1990, 7, 145–
162.
6. Williams, S. J.; Withers, S. G. Carbohydr. Res. 2000, 327,
27–46.
´
´
´
ˇ
´
7. Fialova, P.; Weignerova, L.; Rauvolfova, J.; Prikrylova,
´
V.; Pisˇvejcova, A.; Ettrich, R.; Kuzma, M.; Sedmera, P.;
Krˇen, V. Tetrahedron 2004, 60, 693–701.
8. Tauss, A.; Greimel, P.; Rupitz, K.; Steiner, A. J.; Stutz, A.
¨
E.; Withers, S. G.; Wrodnigg, T. M. Tetrahedron: Asym-
metry 2005, 16, 159–165.
9. Saha, B. C. Biotechnol. Adv. 2000, 18, 403–423.
10. Ho¨vel, K.; Shallom, D.; Niefind, K.; Belakhov, V.;
Shoham, G.; Baasov, T.; Shoham, Y.; Schomburg, D.
EMBO J. 2003, 22, 4922–4932.
11. Miletti, L. C.; Marino, K.; Marino, C.; Colli, W.; Alves,
M. J. M.; de Lederkremer, R. M. Mol. Biochem. Parasitol.
2003, 127, 85–88.
12. Wallis, G. L. F.; Hemming, F. W.; Peberdy, J. F. Biochim.
Biophys. Acta 2001, 1525, 19–28.
13
H-5b); C NMR (D2O, 125.76): d 161.20 (C-10),
142.10 (C-40), 126.10 (C-30, C-50), 116.43 (C-20, C-60),
107.63 (C-1b), 103.99 (C-1a), 88.06 (C-2a), 81.80 (C-
2b), 81.58 (C-3b), 79.78 (C-3a), 79.37 (C-4b), 79.08 (C-
4a), 17.23 (C-5b), 16.93 (C-5a); HRMS (ESI+) for
C16H21NO9Na: m/z [M + Na]+ calcd 394.1114, found:
394.1116.
´
13. Remond, C.; Ferchichi, M.; Aubry, N.; Plantier-Royon,
4.17. p-Nitrophenyl 5-deoxy-a-L-arabinofuranosyl-
(1!3)-5-deoxy-a-L-arabinofuranoside (17)
R.; Portella, C.; O’Donohue, M. J. Tetrahedron Lett. 2002,
43, 9653–9655.
´
14. Remond, C.; Plantier-Royon, R.; Aubry, N.; Maes, E.;
(ꢂ1 mg accumulated from a few preparative plates)
HPTLC (CH2Cl2–MeOH, 4:1) Rf = 0.49; 1H NMR
Bliard, C.; O’Donohue, M. J. Carbohydr. Res. 2004, 330,
2019–2025.