Biocatalytic separation of a-hydroxyphosphonates with lipase of burkholderia cepacia

Full text of DOI:10.1134/S1070363210080244

ISSN 1070-3632, Russian Journal of General Chemistry, 2010, Vol. 80, No. 8, pp. 1718–1719. © Pleiades Publishing, Ltd., 2010.
Original Russian Text © О.О. Kolodyazhnaya, О.I. Kolodyazhnyi, 2010, published in Zhurnal Obshchei Khimii, 2010, Vol. 80, No. 8, pp. 1397–1398.
LETTERS
TO THE EDITOR
Biocatalytic Separation of α-Hydroxyphosphonates
with Lipase of Burkholderia cepacia
О. О. Kolodyazhnaya and О. I. Kolodyazhnyi
Institute of Bioorganic Chemistry and Petroleum Chemistry, National Academy of Sciences of Ukraine,
ul. Murmanskaya 1, Kiev, 02094 Ukraine
fax: 38(044)573-2555
e-mail: olegkol321@rambler.ru
Received January 12, 2010
DOI: 10.1134/S1070363210080244
In recent years enzymes are with increasing
frequency used to obtain the enantiomerically pure
organophosphorus compounds [1–4]. In this work we
found that lipase of Burkholderia cepacia (BCL)
immobilized on diatomite [5, 6] is effective bio-
catalyst, which makes possible a kinetic separation of
racemic α-hydroxyphosphonates into enatiomers. In
the presence of this lipase vinyl acetate esterifies only
(S)-enantiomer of racemic α-hydroxyphosphonates Iа–
Ic to give a mixture containing 50% of α-acylphos-
phonate (S)-IIа–IIc and 50% of hydroxyphosphonate
(R)-Iа–Ic. The latter can be separated by column
chromatography. It was established that the esterifica-
tion rate and optical purity of products little depend on
the solvent (THF, toluene, vinyl acetate), but they
strongly depend on temperature and lipase excess.
When temperature is raised to 40 or 60°С, the reaction
rate enhances approximately 1.5 and 2 times, respect-
tively. The time it takes to reach 50% esterification of
α-hydroxyphosphonates, i.e. to the complete esterifica-
tion of (S)-enantiomer, decreases as the biocatalyst
amount increases. For example, for compound Iа in THF
at 20°С 50% conversion time (depending on the weight
ratio hydroxyphosphonate:lipase) was changed as follows:
48 h (1:0.05), 36 h (1:0.1), 24 h (1:0.2), 16 h (1:0.5), 12 h
(1:1). The rate of 50% conversion for compounds Iа–
Ic became slower as increased the length of the alkyl
substituent R: 12 h (R = Мe) and 60 h (R = Pr) in THF
at 25°С, 300 h (C5H11) in THF at 45°С.
Racemic hydroxyphosphonates were obtained by
the earlier reported method using the reaction of
triethylphosphite with aldehydes in the presence of
pyridinium bromide [7].
OAc
R
EtO
R
R
EtO
EtO
R
H
[C5H5NH]+Br
P
EtO
O
P
P
H
(EtO)3P +
+
EtO
EtO
H
H
−EtBr, −C5H5N
BCL
OH
O
OH
(R+S)-Iа−Ic
R = Me (a), Pr (b), n-С5H11 (c).
OAc
(S)-IIа−IIc
O
O
(R)-Iа−Ic
Optical purity of compounds (S)-IIа–IIc and (R)-
Iа–Ic was established by means of 31Р NMR
spectrometry in a chiral solvating agent (cinchonidine)
medium [8], which showed high (≥ 99%) optical purity
of the separated enantiomers. Optical purity and
absolute configuration of the compounds were
confirmed by derivatization with Mosher’s acid [9].
Furthermore, we compared the optical rotation angles
for the obtained hydroxyphosphonates Iа and Ib with
published data [4].
(R)-Diethyl 1-hydroxyethylphosphonate (Iа). To
a solution of 1.5 g (0.076 mol) of racemic (S/R)-Ia in
3 ml of THF and 3 ml of vinyl acetate was added
1718

BIOCATALYTIC SEPARATION OF Α-HYDROXYPHOSPHONATES
1719
0.15 g of lipase. The mixture was stirred for 48 h at
room temperature. The lipase was filtered off, the
solution was concentrated, and the residue was
subjected to chromatography on a silica gel to form
two fractions, one of which (Rf 0.25, hexane–acetone
2:1) was optically pure alcohol (R)-Ia. Yield 48%, bp
J 4.5, J 9); 4.1.5 m (4H, CH3CH2O). 13C NMR spec-
trum (CDCl3), δ, ppm: 14.01 (CH3); 16.20 d
(CH3CH2O, J 6.0); 16.30 d (CH3CH2O, J 5.8); 22.6
(CH2); 25.99 d (CH2, J 10); 30.3 (CH2); 30.5 (CH2);
61.9 d (CH2O, J 6.0); 62.62 d (CH2O, J 7.1); 66.9 d
(PCH, J 165). Found, %: P 13.12. C10H23O4P. Calcu-
lated, %: P 13.00.
85°С (0.1 mm Hg), [α]D20 –7.0 (c 3, CHCl3). Н NMR
1
spectrum (CDCl3), δ, ppm (J, Hz): 1.36 t (6H,
CH3CH2O, J 6); 1.45 d.d (3H, CH3CP, J 7, J 18); 3.61
br (1H, OH); 4.05 m (1Н, СНР); 4.19 m (4H,
CH3CH2O). 13C NMR spectrum (CDCl3), δ, ppm: 16.2
d (СН3, J 6); 17.28 s (СН3); 61.63 d (СН2О, J 6); 62.4
d (СН2О, J 6); 62.8 d (РС, J 162.5). 31Р NMR
spectrum (CDCl3),δP, ppm:25.8 [2, 4].
(S)-Diethyl 1-acetoxyhexylphosphonate (IIc).
Yield 40%, [α]D20 +30.0 (c 2, CHCl3). 1Н NMR
spectrum (CDCl3), δ, ppm (J, Hz): 0.85 t (3H, CH3,
J 7); 1.26 t (3H, CH3CH2O, J 7); 1.28 t (3H,
CH3CH2O, J 7); 1.2–1.5 m (6H, CH2); 1.77 m (2H,
CH2); 2.06 s (3H, COCH3); 4.1 m (4H, CH3CH2O);
5.20 m (1H, PCH). 13C NMR spectrum (CDCl3), δ,
ppm: 13.9 (CH3); 16.3 d (CH3CH2, J 6.0); 16.4 d
(CH3CH2, J 6); 20.9 (COCH3); 22.8 (CH2); 25.9
(CH2); 29.3 (CH2); 31.0 (CH2); 62.0 d (CH2O, J 6.5);
62.5 d (CH2O, J 7.0); 71.0 d (PCH, J 165); 170.0
(C=O, J 6). Found, %: C 51.21; H 9.00; P 10.92.
C12H25O5P. Calculated, %: C 51.42; H 8.99; P 11.05.
Other fraction (Rf 0.55, hexane–acetone 2:1) was
(S)-diethyl 1-acetyloxyethylphosphonate (IIа). Yield
49%, [α]D20 +25.0 (с 2, CHCl3). Н NMR spectrum
1
(CDCl3), δ, ppm (J, Hz): 1.3 t (6H, CH3СН2O, J 6);
1.45 d.d (3H, CH3CНP, J 7, J 17); 2.1 s (3H, CH3CO);
4.15 m (4H, CH2O); 5.1 d.q (1H, CHP, J 7, J 8). 31Р
NMR spectrum (CDCl3),δP, ppm: 21.1 [2].
The NMR spectra were recorded on a Varian-300
instrument relative to internal TMS (1Н, 13С) and 85%
Н3РО4 in D2O (31Р).
(R)-Diethyl 1-hydroxybutylphosphonate (Ib).
Yield 47%, [α]D20 –25.0 (с 3, CHCl3), bp 120 (0.1 mm
Hg). 1Н NMR spectrum (CDCl3), δ, ppm (J, Hz): 0.9 t
(3H, CH3, J 7); 1.1–1.3 m (2H, CH2); 1.36 t (3H,
CH3CH2О, J 7); 1.38 t (3H, CH3CH2О, J 7); 1.6 m
(2H, CH2); 3.3 br (1H, OH); 3.8 m (1H, PCH); 4.2 m
(4H, OCH2). 13C NMR spectrum (CDCl3), δ, ppm:
13.6 s (CH3); 16.5 s (CH3CH2O, J 8.5); 18.5 d (CH2,
J 7.5); 33.5 d (CH2, J 5); 62 d (OCH2, J 7); 70.5 d (PC,
J 165). 31Р NMR spectrum (CDCl3), δP: 26.
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RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 80 No. 8 2010