Enzymatic resolution of racemic secondary alcohols by lipase B from Candida antarctica

Article abstract of DOI:10.1007/s11746-000-0161-y

Chiral intermediates S-(+)-2-pentanol and S-(+)-2-heptanol were prepared by a lipase-catalyzed enzymatic resolution process. Among various lipases evaluated for the stereoselective acylation of racemic alcohols, lipase B from Candida antarctica catalyzed the acylation of the undesired enantiomer of racemic alcohols leaving the desired S-(+)-alcohols unreacted. A reaction yield of 43-45% and an enantiomeric excess (e.e.) of >99% were obtained for S-(+)-2-pentanol or S-(+)-2-heptanol when the reaction was carried out using vinyl acetate or succinic anhydride as acylating agent. In an alternative process, an enantioselective hydrolysis of 2-pentyl acetate was demonstrated using lipase B giving S-(+)-2-pentyl acetate and R-(-)-2-pentanol. A reaction yield of 45% and an e.e. of 98.6% were obtained for S-(+)-2-pentyl acetate.

Full text of DOI:10.1007/s11746-000-0161-y

Enzymatic Resolution of Racemic Secondary Alcohols
by Lipase B from Candida antarctica1
Ramesh N. Patel,* Amit Banerjee, Venkata Nanduri, Animesh Goswami,
and F.T. Comezoglu
Department of Enzyme Technology, Process Research, Bristol-Myers Squibb Pharmaceutical Research Institute,
a solvent in enzyme-catalyzed reactions. We are also describ-
ing acetylation of secondary alcohols using vinyl acetate and
the hydrolysis of 2-pentyl acetate.
New Brunswick, New Jersey 08903
ABSTRACT:—Chiral intermediates S-(+)-2-pentanol and S-(+)-
2-heptanol were prepared by a lipase-catalyzed enzymatic res-
olution process. Among various lipases evaluated for the stere-
oselective acylation of racemic alcohols, lipase B from Candida
antarctica catalyzed the acylation of the undesired enantiomer
of racemic alcohols leaving the desired S-(+)-alcohols unre-
acted. A reaction yield of 43–45% and an enantiomeric excess
(e.e.) of >99% were obtained for S-(+)-2-pentanol or S-(+)-2-
heptanol when the reaction was carried out using vinyl acetate
or succnic anhydride as acylating agent. In an alternative
process, an enantioselective hydrolysis of 2-pentyl acetate was
demonstrated using lipase B giving S-(+)-2-pentyl acetate and
R-(–)-2-pentanol. A reaction yield of 45% and an e.e. of 98.6%
were obtained for S-(+)-2-pentyl acetate.
MATERIALS AND METHODS
Materials. Racemic 2-pentanol, racemic 2-heptanol, S-(+)-2-
pentanol, S-(+)-2-heptanol, R-(–)-2-pentanol, R-(−)-2-heptanol,
and succinic anhydride were purchased from Aldrich Chemi-
cals (Milwaukee, WI). Candida antarctica lipase B (Chirazyme
L-2) and other lipases (lipases L-3–L-8) were obtained from
Boehringer Mannheim (Indianapolis, IN); lipase PS-30, lipase
FAP-15, lipase AK, lipase MAP-10, and lipase R were pur-
chased from Amano International Enzyme Company (Troy,
VA); porcine pancreatic lipase (PPL) was purchased from
Sigma Chemicals (St. Louis, MO); and Lipomod 200 I was pur-
chased from Biocatalysts Ltd., (Pontypridd, Wales, UK).
Analytical methods. Gas chromatography (GC) was per-
formed on a Hewlett-Packard 5890 gas chromatograph (Palo
Alto, CA) by using a cross-bonded 100% polydimethyl-poly-
siloxane capillary GC column (15 m × 0.32 µm, Rtx-1, Restek
Corporation, Bellefonte, PA) with helium as carrier gas at 8.5
mL/min and a flame-ionization detector.
Method 1: The injector temperature was 150°C and the de-
tector temperature was 200°C. The initial column temperature of
30°C was maintained for 10 min and then increased at the rate of
5°C/min to 50°C and kept at 50°C for an additional 10 min.
Method 2: The injector temperature was 200°C and the de-
tector temperature was 250°C. The initial column tempera-
ture of 100°C was maintained for 10 min and then increased
at the rate of 10°C/min to 200°C and kept at 200°C for an ad-
ditional 2 min.
Paper no. J9652 in JAOCS 77, 1015–1019 (October 2000).
KEY WORDS:—Anti-Alzheimer drugs, Candida antarctica, chiral
intermediates, enzymatic resolution, lipase, secondary alcohols.
The current interest in enzymatic production of chiral com-
pounds lies in the preparation of intermediates for pharma-
ceutical synthesis (1–10). S-(+)-2-pentanol is a key chiral in-
termediate required for synthesis of anti-Alzheimer drugs that
inhibit β-amyloid peptide release and/or its synthesis (11–13).
Chiral secondary alcohols have been pepared by microbial
reduction of ketones (14–18). Resolution of secondary alco-
hols by lipase- and acylase-catalyzed reactions has been
demonstrated (19–27), but most of the reports were on the
resolution of compounds having bulky groups such as phenyl
groups. In this report we describe an enzymatic process for
the resolution of the aliphatic compounds such as racemic 2-
pentanol and 2-heptanol by lipase B from Candida antarc-
tica. By using succinic anhydride as acylating agent, the un-
reacted desired chiral alcohols can be easily separated from
the product. We have scaled up the resolution process and
have used substrates (racemic 2-pentanol and 2-heptanol) as
Method 3: Separation of enantiomers of racemic 2-pen-
tanol and 2-heptanol was carried out by GC. Samples from
reaction mixtures were extracted with ethyl acetate and dried
over anhydrous magnesium sulfate. Analyses of samples
were carried out using a Hewlett-Packard 5800 GC with
flame-ionization detector. Astec Chiraldex G-TA (Whippany,
NJ) (gamma cyclodextrin, 20 m × 0.25 mm × 0.125 mm
thickness) was used at 28°C (for 2-pentanol) or at 44°C (for
2-heptanol). Helium was used as carrier gas at 22 mL/min.
The injector temperature was 150°C, and the detector temper-
1This work was presented at the Biocatalysis Symposium in April 2000, held
at the 91st Annual Meeting and Expo of the American Oil Chemists’ Soci-
ety, San Diego, CA.
*To whom correspondence should be addressed at Department of Enzyme
Technology, Process Research, Bristol-Myers Squibb Pharmaceutical Re-
search Institute, P.O. Box 191, New Brunswick, NJ 08903.
E-mail: patelr@bms.com
Copyright © 2000 by AOCS Press
1015
JAOCS, Vol. 77, no. 10 (2000)

1016
R.N. PATEL ET AL.
ature was 200°C. Initial oven temperature was 28°C for 15 succinic anhydride: Isolation of hemisuccinate. Enzymatic
min, and then increased at the rate of 5°C/min to 50°C and acylation was carried out with neat racemic alcohol, 0.68
held at 50°C for 5 min for 2-pentanol analysis. Under the molar equivalent succinic anhydride, and 13 mg lipase B/g of
above conditions the retention times for S-2-pentanol and R- racemic alcohol. The reaction was carried out at 35°C and 150
2-pentanol were 10.6 and 11.6 min, respectively. The reten- rpm for 48 h. The reaction was stopped by filtration. The
tion times for S-2-heptanol and R-2-heptanol were 11.2 and hemisuccinate was isolated as follows.
11.8 min, respectively.
A portion of the filtered reaction mixture was dissolved in
The nuclear magnetic resonance (NMR) spectra were MTBE (5 vol). The MTBE solution was extracted with 5%
recorded in CDCl3 solution using a JEOL (Tokyo, Japan) NaHCO3 to extract the hemisuccinate in base. The MTBE ex-
NMR spectrophotometer operating at 399.8 MHz for 1H and tract contained unreacted alcohol. The 5% NaHCO3 extract
100.5 MHz for 13C.
was washed with MTBE until free from alcohol. The
Lipase screening for resolution of 2-pentanol. Screening of hemisuccinate was liberated by slow acidification of the 5%
various lipases for the resolution of racemic 2-pentanol was car- NaHCO3 layer with 1 N HCl to pH 4. The hemisuccinate was
ried out in 10 mL of hexane containing 50 mg of racemic 2-pen- extracted in MTBE from the acidic solution. The final MTBE
tanol, 200 mg of vinyl acetate, and 50 mg of enzyme. Reactions solution was washed with 5% NaCl until neutral pH was at-
were carried out at 30°C, 150 rpm for 72 h on a rotary shaker. tained, dried over anhydrous Na2SO4, filtered, and the solvent
Samples were analyzed by GC method 1.
was removed on a water bath maintained at 70–80°C. The
Preparative scale acetylations of racemic 2-pentanol were hemisuccinates obtained were analyzed by GC method 2 as
carried out in heptane. The reaction mixture in 1 L of heptane described in analytical methods. The retention times for 2-
contained 100 g of racemic 2-pentanol, 1.02 mole equivalent pentyl hemisuccinate and 2-heptyl hemisuccinate were 5.9
of vinyl acetate, and 1 g of lipase B. The reaction was carried and 8.7 min, respectively.
out at 35°C and 150 rpm.
2-Pentyl hemisuccinate. 1H NMR: δ (ppm) 0.81 (3H, t, J
Preparative scale acylation of racemic 2-pentanol was also = 7 Hz, 5-CH3), 1.11 (3H, d, J = 6 Hz, 1-CH3), 1.2–1.5 (4H,
carried out in racemic 2-pentanol or 2-heptanol as a solvent. m, 3- and 4-CH2), 2.52 (4H, AB quartet, 2′ and 3′-CH2), 4.84
The reaction mixture contained 1 kg of racemic alcohol, 0.68 (1H, m, 2-CH). 13C NMR: δ (ppm) 14.52 (C-5), 19.21 (C-4),
mole equivalent of succinic anhydride, and 13 g of lipase B. 20.49 (C-1), 29.57 (C-2′), 29.86 (C-3′), 38.49 (C-3), 71.54
The reaction was carried out at 38°C and 150 rpm.
(C-2), 171.31 (C-1′), 176.6 (C-4′).
Preparation of RS-acetate. To a solution of RS-alcohol (2
2-Heptyl hemisuccinate. 1H NMR: δ (ppm) 0.86 (3H, t, J =
mL) in pyridine (5 mL), acetic anhydride (5 mL) was added 7 Hz, 7-CH3), 1.18 (3H, d, J = 6 Hz, 1-CH3), 1.2–1.6 (8H, m,
and the mixture kept at room temperature for 16 h. The mix- 3,4,5,6-CH2), 2.6 (4H, AB quartet, 2′ and 3′ CH2), 4.91 (1H, m,
ture was poured in ice cold water and extracted with methyl 2-CH). 13C NMR: δ (ppm) 14.75 (C-7), 20.59 (C-1), 23.21
tertiary butyl ether (MTBE, 3 × 20 mL). The MTBE extract (C-6), 25.67 (C-4), 29.7 (C-2′), 29.86 (C-3′), 32.22 (C-5), 36.39
was washed with water (2 × 10 mL), 1 N HCl (2 × 10 mL) (C-3), 72.02 (C-2), 171.27 (C-1′), 177.84 (C-4′).
and finally with water (3 × 10 mL), dried over Na2SO4, fil-
Isolation of S-(+)-2-pentanol. A neat reaction mixture
tered, and solvent was removed on a water bath maintained at from acylation of racemic 2-pentanol (1.2 L) containing 399
70–80°C. The retention times of the acetates and alcohols by g (4.526 moles, 44.3 M%) of S-(+)-2-pentanol was used to
GC method 1 were 2-pentanol 1.7 min, 2-pentyl acetate 5.6 develop the recovery process. A portion of the reaction mix-
min, 2-heptanol 9.5 min, and 2-heptyl acetate 19.9 min.
ture (600 mL) containing S-(+)-2-pentanol (193.3 g, 2.193
2-Pentyl acetate. 1H NMR: δ (ppm) 0.83 (3H, t, J = 7 Hz, moles) was cooled to –12–14°C and the precipitates were re-
5-CH3), 1.12 (3H, d, J = 6 Hz, 1-CH3), 1.2–1.5 (4H, m, 3- and moved by filtration. The clarified crude oil was distilled, with
4-CH2), 1.93 (3H, s, 2′-CH3), 4.83 (1H, m, 2-CH). 13C NMR: the S-(+)-2-pentanol distilling at 51 to 67°C under vacuum
δ (ppm) 14.55 (C-5), 19.29 (C-2′), 20.57 (C-1), 21.92 (C-4), (20–25 mm Hg) to yield 165 g of product [1.866 moles, 98%
38.57 (C-3), 70.96 (C-2), 170.11 (C-1′).
enantiomeric excess (e.e.), GC homogeneity index 99.5%].
2-Heptyl acetate. 1H NMR: δ (ppm) 0.82 (3H, t, J = 7 Hz,
Isolation of S-(+)-2-heptanol. A neat reaction mixture from
7-CH3), 1.14 (3H, d, J = 6 Hz, 1-CH3), 1.2–1.6 (8H, m, acylation of racemic 2-heptanol (67.5 mL) containing 22 g of
3,4,5,6-CH2), 4.84 (1H, m, 2-CH). 13C NMR: δ (ppm) 14.68 S-(+)-2-heptanol was used to develop a recovery process. A por-
(C-7), 20.60 (C-2′), 21.98 (C-1), 23.19 (C-6), 25.70 (C-4), tion of the reaction mixture (62 mL) containing S-(+)-2-heptanol
32.21 (C-3), 36.42 (C-5), 71.26 (C-2), 170.16 (C-1′).
(20.13 g, 0.173 moles) was cooled to –12–14°C, and the precip-
Enzymatic acetylation of 2-pentanol and 2-heptanol. Identifi- itates were removed by filtration. The clarified crude oil was dis-
cation of acetate. Enzymatic acetylation was carried out with tilled, with the S-(+)-2-heptanol distilling at 60 to 62°C under
500 mg of RS-alcohol in 10 mL heptane, 1.02 mole equivalent vacuum (6.0 mm Hg) to yield 17.17 g of product (0.148 moles,
of vinyl acetate, and 5 mg of lipase B. The reaction was carried 98% e.e., GC homogeneity index 98.3%).
out at 35°C and 150 rpm for 4 h. The reaction was stopped by
filtration of the enzyme. The enzymatic reaction products were
analyzed by GC method 1 as described above.
RESULTS AND DISCUSSION
Enzymatic acylation of 2-pentanol and 2-heptanol with Commercially available lipases from C. rugosa (lipase L-3),
JAOCS, Vol. 77, no. 10 (2000)

RESOLUTION OF SECONDARY ALCOHOLS
1017
TABLE 1
Screening of Enzymes for Resolution of Racemic 2-Pentanola
2-Pentanol
(g/L)
2-Pentylacetate
(g/L)
Yield of 2-pentanol
(%)
e.e. of S-2-Pentanol
Enzymes
(%)
Candida antarctica lipase B (L-2)
C. rugosa lipase L-3
BM lipase L-7
BM lipase L-8
PPL
Lipomod 200 I
Lipase PS-30
Lipase FAP-15
Lipase AK
2.48
1.06
3.65
5
3.5
5.6
1.97
0
49.6
21.2
73
100
100
51.4
77.8
100
98
99
92
30
00
00
00
20
00
00
—
5
0
2.57
3.89
5
4.9
0
3.4
1.56
0
0.2
7.5
Lipase MAP-10
0
aReaction mixture in 10 mL of hexane contained 50 mg of racemic 2-pentanol, 200 mg of vinyl acetate, and 50 mg of
enzyme as indicated. Reactions were carried out on a rotary shaker at 150 rpm for 72 h. Suppliers: L-2, L-3, L-7, L-8
(Boehringer Mannheim, Indianapolis, IN); PS-30, FAP-15, AK, MAP-10 (Amano International Enzyme Co., Troy, VA),
porcine pancreatic lipase (Sigma Chemical, St. Louis, MO); Lepomod 200 I (Biocatalysts Ltd., Pontypridd, Wales, United
Kingdom). Abbreviations: e.e., enantiomeric excess; PPL, porcine pancreatic lipase.
C. antarctica (lipase B), Penicillium sp. (lipase R), Mucor sp. tanol were carried out using racemic 2-pentanol as solvent as
(lipase MAP-10), porcine pancreatic lipase, Pseudomonas well as substrate. A reaction yield of 43 M% (theoretical maxi-
cepacia (lipase PS-30), Pseudomonas sp. (lipase AK), Geo- mum 50%) and e.e. of 98.8–99.3% were obtained for S-(+)-2-
trichum candidum (lipase G), and Boehringer Mannheim (BM) pentanol. Results from three preparative batches are as shown
lipases L-4, L-5, L-6, L-7, and L-8 were screened for the stere- in Table 3. The E value of 48 was obtained. From the reaction
oselective acetylation of racemic 2-pentanol. Candida antarc- mixture S-(+)-2-pentanol was isolated in overall 36% yield (the-
tica lipase B and C. rugosa lipase efficiently catalyzed the enan- oretical maximum 50%).
tioselective acetylation. The reaction yields and enantiomeric
As described above, the resolution of 2-heptanol was also
excess (e.e.) of S-(+)-2-pentanol obtained with various enzymes carried out using lipase B. A reaction yield of 44 M% (theo-
are shown in Table 1. A reaction yield of 49.6% and an e.e. of retical maximum 50%) and e.e. of 99.3% were obtained for
99% were obtained using C. antarctica lipase B. BM lipases L- S-(+)-2-heptanol. Results from two preparative batches are as
4, L-5, L-6, lipase MAP-10, and lipase R acetylated both enan- shown in Table 4. The E value of 49 was obtained. From the
tiomers of racemic 2-pentanol. BM lipase L-8, immobilized reaction mixture S-(+)-2-heptanol was isolated in overall 40%
Lipomod-200, lipase FAP-15, and lipase AK were inactive in yield (theoretical maximum 50%).
the acetylation reaction. Lower reaction yields and e.e. of S-(+)-
2-pentanol were obtained when other enzymes were used.
To prepare R-(–)-2-pentanol, hydrolysis of racemic 2-pentyl
acetate was carried out using lipase B. S-(+)-2-Pentyl acetate
Since the R-isomer of racemic 2-pentanol is the substrate was obtained in 45% yield with an e.e. of 98.6%. R-(–)-2-pen-
for C. antarctica lipase B catalyzed acylations and the desired tanol was obtained in 48 M% yield with an e.e. of 96%. The E
S-2-pentanol remained unreacted in the reaction mixture, we value of 47 was obtained. The kinetics of the reaction are shown
concentrated our efforts only on the isolation of the desired in Table 5.
S-(+)-2-pentanol. The formation of 2-pentyl acetate by acety-
Various research groups have used different solvents and dif-
lation of racemic 2-pentanol with vinyl acetate was confirmed ferent acylating agents to resolve a variety of complex secondary
by GC comparison with the corresponding acetates prepared alcohols using lipases. Most of the reports were on the resolu-
by conventional chemical acetylation. Preparative scale en- tion of compounds having bulky group such as phenyl. Ema
zymatic acetylation of racemic 2-pentanol was carried out in
heptane. Kinetics of the acetylation reaction are shown in
TABLE 2
Table 2. At the end of the reaction, 44.5 g of S-(+)-2-pentanol
was obtained with an e.e. of 98%. The enantiomeric ratio E
value (28) of 37 was obtained.
Preparative Scale Enzymatic Acetylation of Racemic 2-Pentanol
Using Lipase B from Candida antarcticaa
Reaction time
(h)
2-Pentanol
(g/L)
e.e. of S-2-Pentanol
Among acylating agents tested, succinic anhydride was
found to be the best choice due to easy recovery of S-(+)-2-pen-
tanol at the end of the reaction (Scheme 1). The products 2-
pentyl and 2-heptyl hemisuccinates were isolated from a por-
tion of the enzymatic reaction mixture, and their structures were
confirmed by NMR. Since the desired (S)-alcohols remained in
the reaction mixture, we focused only on the isolation of the de-
sired (S)-alcohols. The enzymatic acylation reactions of 2-pen-
(%)
0
2
4
6
100
70
56
00
32
60
44.52
98
aReaction mixture in 1 L of heptane contained 100 g of racemic 2-pentanol,
1.02 mole equivalent of vinyl acetate, and 1 g of lipase B from C. antarctica.The
reaction was carried out at 35°C and 150 rpm. For supplier and abbreviations
see Table 1.
JAOCS, Vol. 77, no. 10 (2000)

1018
R.N. PATEL ET AL.
Racemic 2-pentanol
R-(–)-2-Pentyl hemisuccinate
Succinic anhydride
Racemic 2-heptanol
Succinic anhydride
S-(+)-2-Heptanol
R-(–)-2-Heptyl hemisuccinate
SCHEME 1
et al. (22) has demonstrated the resolution of a large secondary using diketene as acylating agent in isopropyl ether. 1-Phenyl-
alcohol, 5-[4-(1-hydroxyethyl)phenyl]-10,15,20-triphenylpor- ethanol and a variety of secondary alcohols were resolved by
phyrin by lipase-catalyzed transesterification using vinyl acetate. PPL-catalyzed transesterification reactions in alkylcarbonate as
Lipase PS-30 from P. cepacia and C. antarctica lipase B in dry solvent. Addition of molecular sieves (4Å) in the reaction mix-
diisopropyl ether as a solvent demonstrated high selectivity and ture improved the e.e. of chiral alcohols (27).
gave the (S)-alcohol in 98% e.e. 1-Ethoxyvinyl esters were used
Resolution of 3-hydroxy-1-undecyne, 3-hydroxy-1-nonene,
as acyl donors for enzymatic resolution of secondary alcohols 3-nonanol, and 1-chloro-2-octanol was demonstrated by Orre-
using C. rugosa lipase. The 1-ethoxyvinyl esters generate ethyl nius et al. (26) using lipase B from C. antarctica and S-ethyl
acetate as the co-product, which does not affect the enzyme ac- thiooctanoate as acylating agent. Enzymatic resolution of sec-
tivity, whereas the acetaldehyde liberated from the vinyl esters ondary alcohols under substrate racemizing conditions was stud-
inactivates some lipases (21). Bevinakatti et al. (28) resolved ied by Persson et al. (30) using an immobilized lipase from C.
sterically hindered secondary alcohols by lipase-catalyzed enan- antarctica in the presence of a ruthenium catalyst. A specifically
tioselective transesterification of their O-acetyl esters with pri- designed acyl donor, 4-chlorophenyl acetate, was used with
mary alcohols in diisopropyl ether. Mandelic acid esters, man- toluene or hexane as a solvent. With this process, a variety of
delonitrile, 2-chloro-1-phenylethanol, and pentolactone were re- secondary alcohols were converted to the corresponding enan-
solved using C. rugosa lipase. (S)-Propanolol was prepared by tiomerically pure acetates with >99% e.e.
lipoprotein lipase-catalyzed resolution of racemic alcohol.
In this paper we have reported the lipase-catalyzed resolu-
XAD-8 immobilized enzyme was used in tert-butyl ether as sol- tion of aliphatic compounds such as secondary alcohols lack-
vent with vinyl acetate as acylating agent (29). Suginaka et al. ing bulky groups. In the resolution process, substrates were
(25) have demonstrated the resolution of 1-(1-naphthyl)ethanol, used as solvent, giving about 450 g of resolved (S)-secondary
1-(2-naphthyl)ethanol, 1-phenyl-2-propanol, and 1,2,3,4- alcohol from a 1-L preparative batch. Succinic anhydride was
tetrahydro-1-naphthol by lipase-catalyzed acylation reaction used as an acylating agent, providing easy recovery of the chi-
ral alcohol at the end of the reaction.
TABLE 3
Enzymatic Acylation of Racemic 2-Pentanol Using Succinic
TABLE 4
Anhydride and Lipase B from Candida antarcticaa
Enzymatic Acylation of Racemic 2-Heptanol Using Succinic
Anhydride and Lipase B from Candida antarcticaa
2-Pentanol
input
2-Pentanol
remaining
(g)
e.e. of
S-2-Pentanol
(%)
Batch
number
2-Heptanol
input
2-Heptanol
remaining
(g)
e.e. of
S-2-Heptanol
(%)
(g)
Batch
number
(g)
132
133
136
500
500
900
210
222
400
99.2
98.8
99.3
140
141
100
500
43
222
99.3
99.4
aReaction mixture contained racemic 2-pentanol as solvent and substrate,
0.68 mole equivalent of succinic anhydride, and 13 g of lipase B per kg of
substrate input. The reaction was carried out at 38°C and 150 rpm. For sup-
plier and abbreviations see Table 1.
aReaction mixture contained racemic 2-pentanol as solvent and substrate,
0.68 mole equivalent of succinic anhydride, and 13 g of lipase B per kg of
substrate input. The reaction was carried out at 38°C and 150 rpm. For sup-
plier and abbreviations see Table 1.
JAOCS, Vol. 77, no. 10 (2000)

RESOLUTION OF SECONDARY ALCOHOLS
1019
16. Adlercreutz, P., Asymmetric Reduction of Ketones with En-
zymes from Acetic Acid Bacteria, Biotechnol. Lett. 13:229–234
(1991).
TABLE 5
Enantioselective Enzymatic Hydrolysis of 2-Pentyl Acetate by
Lipase B from Candida antarcticaa
17. Trincone, A., L. Lama, V. Lanzotti, B. Nicolaus, M. DeRosa, M.
Rossi, and A. Gambacorta, Asymmetric Reduction of Ketones with
Resting Cells of Sulfolobus solfataricus, Biotech. Bioeng. 35:
559–564 (1990).
Reaction
time
(mm)
2-Pentyl
acetate
(g/L)
e.e. of S-2-
pentanol acetate
(%)
2-Pentanol
(g)
18. Patel, R.N., C.T. Hou, A.I. Laskin, and P. Derelanko, Microbial
Production of Methylketones: Properties of a Purified Secondary
Alcohol Dehydrogenase from Pichia sp., J. Appl. Biochem. 3:
218–232 (1981).
19. Salzar, L., J.L. Bermudez, C. Ramirez, E.F. Llama, and J.V. Sinis-
terra, Resolution of 3-α-Naphthoxy-1,2-propanediol Using Can-
dida antarctica Lipase, Tetrahedron: Asymmetry 10:3507–3514
(1999).
20. Bidjou, C., and L. Aribi-Zouioueche, Kinetic Resolution of Sec-
ondary Benzyl Derivatives by Transesterification and Enzymic
Hydrolysis, J. Soc. Alger. Chim. 9:261–268 (1999).
21. Kita, Y., Y. Takebe, K. Murata, T. Naka, and S. Akai, Conve-
nient Enzymatic Resolution of Alcohols Using Highly Reactive,
Nonharmful Acyl Donors, 1-Ethoxyvinyl Esters, J. Org. Chem.
65:83–88 (2000).
22. Ema, T., M. Jittani, T. Sakai, and M. Utaka, Lipase-Catalyzed
Kinetic Resolution of Large Secondary Alcohols Having Tetra-
phenylporphyrin, Tetrahedron Lett. 39:6311–6314 (1998).
23. Faraldos, J., E. Arroyo, and B. Herradon, Biocatalysis in Organic
Synthesis. Part 9. Highly Enantioselective Kinetic Resolution of
Secondary Alcohols Catalyzed by Acylase, Synletters 4:
367–370 (1997).
9
15
30
90
150
100.
78.
67.
48.
45.2
0.
15.6
23.
37.
39.
0.
28.
46
83.
98.6
aReaction mixture in 1 L of 50 mM phosphate buffer (pH 7.0) contained 100
g of racemic 2-pentyl acetate and 0.6 g of lipase B. The pH was maintained
at 7.0 with 5 N NaOH during reaction.The reaction was carried out at 32°C,
150 rpm. For abbreviation see Table 1.
REFERENCES
1. Jones, J.B., Enzymes in Organic Synthesis, Tetrahedron 42:
3351–3403 (1986).
2. Crout, D.H.G., S. Davies, R.J. Heath, C.O. Miles, D.R. Rath-
bone, and B.E.P. Swoboda, Application of Hydrolytic and De-
carboxylating Enzymes in Biotransformations, Biocatalysis 9:
1–30 (1994).
3. Davies, H.G., R.H. Green, D.R. Kelly, and S.M. Roberts, Re-
cent Advances in the Generation of Chiral Intermediates Using
Enzymes, Biotechnology 10:129–152 (1990).
4. Csuk, R., and B.I. Glanzer, Baker’s Yeast Mediated Transfor-
mations in Organic Synthesis, Chem. Rev. 96:556–566 (1991).
5. Kamphuis, J., W.H.J. Boesten, Q.B. Broxterman, H.F.M. Hermes,
J.A.M. van Balken, E.M. Meijer, and H.E. Schoemaker, The Pro-
duction and Uses of Optically Pure Natural and Unnatural Amino
Acids, Adv. Biochem. Eng. Biotech. 42:133–186 (1990).
6. Sih, C.J., Q-M. Gu, X. Holdgrun, and K. Harris, Optically-Active
Compounds via Biocatalytic Methods, Chirality 4:91–97 (1992).
7. Santaneillo, E., P. Ferraboschi, P. Grisenti, and A. Manzocchi, The
Biocatalytic Approach to the Preparation of Enantiomerically Pure
Chiral Building Blocks, Chem. Rev. 92:1071–1140 (1992).
8. Margolin, A.L., Enzymes in the Synthesis of Chiral Drugs, En-
zyme Microb. Technol. 15:266–280 (1993).
9. Wong, C-H., and G.M. Whitesides, Enzymes in Synthetic Or-
ganic Chemistry, Tetrahedron Organic Chemistry Series, Perga-
mon, New York, 1994, Vol. 12.
10. Patel, R., Stereoselective Biotransformations in Synthesis of Some
Pharmaceutical Intermediates, Adv. Appl. Microbiol. 43:91–140
(1997).
11. Sauerberg, P., P.H. Olesen, Heterocyclic Compounds and Their
Preparation and Use, U.S. Patent 5,418,240 (1995).
12. Hamilton, G.S., J-H., Li, and J.P. Steiner, Method of Using Neu-
trotrophic Sulfonamide Compounds, U.S. Patent 5,721,256 (1998).
13. Caballa, D., I. Francois, and S. Hodgson, Highlights from Society
For Medicine’s Research Meeting, Sept. 18, 1997, London, DN &
P 10 (8):October 1997.
14. Adlercreutz, P., Novel Biocatalysts for the Asymmetric Reduction
of Ketones: Permeabilized Cells of Gluconobacter oxydans, En-
zyme Microb. Technol. 13:9–14 (1991).
24. Legros, J.-Y., M. Toffano, S.K. Drayton, M. Rivard, and J.-C.
Fiaud, Kinetic Resolution of Secondary Alcohols Mediated by
Rabbit Gastric Lipase, Tetrahedron Lett. 38:1915–1918 (1997).
25. Suginaka, K., Y. Yahashi, and Y. Yamamoto, Highly Selective
Resolution of Secondary Alcohols and Acetoacetates with Li-
pases and Diketene in Organic Media, Tetrahedron: Asymmetry
7:1153–1158 (1996).
26. Orrenius, C., N. Oehrner, D. Rotticci, A. Mattson, K. Hult, and T.
Norin, Candida antarctica Lipase B Catalyzed Kinetic Resolu-
tions: Substrate Structure Requirements for the Preparation of
Enantiomerically Enriched Secondary Alcohols, Ibid. 6:1217–
1220 (1995).
27. Janssen, A.J.M., A.J.H. Klunder, and B. Zwanenburg, Resolu-
tion of Secondary Alcohols by Enzyme-Catalyzed Transesterifi-
cation in Alkyl Carboxylates as the Solvent, Tetrahedron
47:7645–7662 (1991).
28. Bevinakatti, H.S., A.A. Banerji, and R.V. Newadker, Resolution
of Secondary Alcohols Using Lipase in Disopropyl Ether, J.
Org. Chem. 54:2453–2455 (1989).
29. Hsu, S.-H., S.-S. Wu, Y.-F. Wang, and C.-H. Wong, Lipase-Cat-
alyzed Irreversible Transesterification Using Enol Esters:
XAD-8 Immobilized Lipoprotein Lipase-Catalyzed Resolution
of Secondary Alcohols, Tetrahedron Lett. 31:6403–6406
(1990).
30. Persson, B.A., A.L.E. Larsson, M.L. Ray, and J.-E. Backvall,
Ruthenium- and Enzyme-Catalyzed Dynamic Resolution of
Secondary Alcohols, J. Am. Chem. Soc. 121:1645–1650 (1999).
15. Zelinski, T., and M-R. Kula, A Kinetic Study and Application
of a Novel Carbonyl Reductase Isolated from Rhodococcus ery-
thropolis, Bioorg. Med. Chem. 2:421–428 (1994).
[Received June 6, 2000; accepted July 28, 2000]
JAOCS, Vol. 77, no. 10 (2000)