Nitrilase-Catalyzed Selective Hydrolysis of Dinitriles
COMMUNICATIONS
Evaporation of the solvent afforded the crude product, References
which was purified bycolumn chromatographyusing ethyl
acetate/hexane (10/90, v/v) as eluent. The a,b-unsaturated
a-cyanoacetate was isolated as a colorless oil in 95% yield.
[1] P. T. Anastas, J. C. Warner, Green Chemistry: Theory
and Practice, Oxford UniversityPress, New York, 1998,
p 152.
[2] U. T. Bornscheuer, R. J. Kazlauskas, Hydrolases in Or-
ganic Synthesis: Regio- and Stereoselective Biotransfor-
mations, Wiley-VCH, Weinheim, 2006, p 368.
A
solution of the a,b-unsaturated a-cyanoacetate
(
5 mmol) and NaCN (8.5 mmol) in 90% ethanol (15 mL)
was refluxed for 5 h. The resulting dark solution was evapo-
rated, and residue was suspended in water (50 mL) and ex-
tracted with dichloromethane. The organic extract was dried
[
3] M.-X. Wang, Top. Catal. 2005, 35, 117–130.
[4] L. Martinkova, V. Mylerova, Curr. Org. Chem. 2003, 7,
279–1295.
over anhydrous Na SO . Removal of solvents afforded 1-cy-
2
4
anocyclohexaneacetonitrile as a white solid; yield: 0.52 g
1
1
13
(
(
70%), which was characterized by H and C NMR data
[18,30]
[
5] T. Sugai, T. Yamazaki, M. Yokoyama, H. Ohta, Biosci.
Biotechnol. Biochem. 1997, 61, 1419–1427.
see Supporting Information).
[
6] J. A. Chaplin, M. D. Levin, B. Morgan, N. Farid, J. Li,
Z. Zhu, J. McQuaid, L. W. Nicholson, C. A. Rand, M. J.
Burk, Tetrahedron: Asymmetry 2004, 15, 2793–2796.
7] E. C. Hann, A. E. Sigmund, S. K. Fager, F. B. Cooling,
J. E. Gavagan, M. G. Bramucci, S. Chauhan, M. S.
Payne, R. DiCosimo, Tetrahedron 2004, 60, 577–581.
Preparation of 1-Cyanocycloalkaneacetic Acids
The general procedure was followed: 1-Cyanocyclopent-
aneacetonitiles (200 mg, 1.31 mmol) was treated with the ni-
trilase (10 mg, 13.5 U) in potassium phosphate buffer
[
(
50 mL, 100 mM, pH 7.2). The reaction mixture was incubat-
13
[8] G. DeSantis, K. Wong, B. Farwell, K. Chatman, Z. Zhu,
G. Tomlinson, H. Huang, X. Tan, L. Bibbs, P. Chen, K.
Kretz, M. J. Burk, J. Am. Chem. Soc. 2003, 125, 11476–
ed at 308C and the reaction was monitored by C NMR.
When dinitrile was consumed, the mixture was acidified
with 1 N HCl solution to pH ~5. After being saturated with
NaCl, the mixture was extracted with ethyl acetate. The or-
ganic extract was dried over anhydrous sodium sulfate. Re-
moval of solvent provided the desired product, which was
further purified bypreparative TLC using eth ly acetate-
hexane (35/65, v/v) as eluting solvent.
1
1477.
[
9] G. DeSantis, Z. Zhu, W. A. Greenberg, K. Wong, J.
Chaplin, S. R. Hanson, B. Farwell, L. W. Nicholson,
C. L. Rand, D. P. Weiner, D. E. Robertson, M. J. Burk,
J. Am. Chem. Soc. 2002, 124, 9024–9025.
1
-Cyanocyclopentaneacetic acid: Yield: 210 mg (92%);
[10] S. Osswald, H. Wajant, F. Effenberger, Eur. J. Biochem.
1
mp 79–818C; H NMR (400 MHz, CDCl ): d=2.74 (s, 2H),
2002, 269, 680–687.
3
2
.32–2.35 (m, 2H), 1.90–1.93 (m, 2H), 1.7–1.85 (m, 4H);
[11] F. Effenberger, S. Osswald, Synthesis 2001, 1866–1872.
[12] F. Effenberger, S. Osswald, Tetrahedron: Asymmetry
2001, 12, 279–285.
13
C NMR (100.6 MHz, CDCl ): d=175.6, 124.4, 42.3, 39.9,
3
3
9
8.6, 24.4; anal. calcd. for C H NO : C 62.73, H 7.24, N
8
11
2
.14; found: C 62.54, H 7.35, N 8.92.
-Cyanocyclohexaneacetic acid: Yield: 198 mg (88%);
[13] F. Effenberger, S. Osswald, Tetrahedron: Asymmetry
1
2001, 12, 2581–2587.
[16]
1
mp 103–1058C, lit.
CDCl ): d=2.64 (s, 2H), 2.13–2.16 (m, 2H), 1.64–1.78 (m,
5
102–1038C; H NMR (400 MHz,
[14] E. C. Hann, A. E. Sigmund, S. K. Fager, F. B. Cooling,
J. E. Gavagan, A. Ben-Bassat, S. Chauhan, M. S. Payne,
S. M. Hennessey, R. DiCosimo, Adv. Synth. Catal. 2003,
345, 775–782.
[15] J. E. Gavagan, S. K. Fager, R. D. Fallon, P. W. Folsom,
F. E. Herkes, A. Eisenberg, E. C. Hann, R. DiCosimo,
J. Org. Chem. 1998, 63, 4792–4801.
3
1
3
H), 1.35–1.41 (m, 2H), 1.20–1.27 (m, 1H); C NMR
(
100.6 MHz, CDCl ): d=175.1, 122.6, 42.2, 36.6, 35.7, 25.4,
3
23.1; anal. calcd. for C H NO : C 64.65, H 7.84, N 8.38;
9 13 2
found: C 64.56, H 8.01, N 8.21.
-Cyanocycloheptaneacetic acid: In this case, the conver-
sion was not complete even after 48 h as monitored by
1
[
16] R. A. Jennings, D. R. Johnson, R. E. Seamans, J. R.
13
C NMR; 65 mg (33%) of starting material was recovered;
Zeller, (Warner-Lambert Co., USA), US Patent
1
yield: 123 mg (55%); mp 131–1338C; H NMR (400 MHz,
CDCl ): d=2.67 (s, 2H), 2.13–2.17 (m, 2H), 1.70–1.80 (m,
8
d=175.3, 123.6, 44.6, 39.2, 38.2, 28.1, 23.6; anal. calcd. for
C H NO : C 66.27, H 8.34, N 7.73; found: C 66.03, H 8.41,
N 7.58
5
,319,135, 1994.
3
[
[
17] M. P. Burns, J. W. Wong, (Pfizer Products Inc.), World
1
3
H), 1.54 À1.65 (m, 2H); C NMR (100.6 MHz, CDCl ):
3
Patent WO 2004111256, 2004.
18] C. E. Burgos-Lepley, L. R. Thompson, C. O. Kneen,
S. A. Osborne, J. S. Bryans, T. Capiris, N. Suman-Chau-
han, D. J. Dooley, C. M. Donovan, M. J. Field, M. G.
Vartanian, J. J. Kinsora, S. M. Lotarski, A. El-Kattan,
K. Walters, M. Cherukury, C. P. Taylor, D. J. Wustrow,
J. B. Schwarz, Bioorg. Med. Chem. Lett. 2006, 16, 2333–
10
15
2
Acknowledgements
2
336.
[
[
[
[
19] O. Meth-Cohn, M.-X. Wang, J. Chem. Soc., Perkin
The authors thank Southern Methodist University and
Robert. A. Welch Foundation for financial support.
Trans. 1 1997, 3197–3204.
20] C. Mukherjee, D. Zhu, E. R. Biehl, L. Hua, Eur. J. Org.
Chem. 2006, 5238–5242.
21] H. Kakeya, N. Sakai, A. Sano, M. Yokoyama, T. Sugai,
H. Ohta, Chem. Lett. 1991, 1823–1824.
22] M.-X. Wang, C.-S. Liu, J.-S. Li, O. Meth-Cohn, Tetrahe-
dron Lett. 2000, 41, 8549–8552.
Adv. Synth. Catal. 2007, 349, 1667 – 1670
ꢀ 2007 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
1669