Enzyme-mediated enantioselective hydrolysis of dicarboxylic acid monoesters

Article abstract of DOI:10.2174/157017812803521108

The enzyme-mediated highly enantioselective hydrolysis of dicarboxylic acid monoesters was investigated. The racemic substrates, which were prepared by coupling of the corresponding alcohols with dicarboxylic anhydrides, were enantioselectively hydrolyzed by lipase from Candida antarctica (Novozym 435) in a buffer at 30 °C. The products were easily separated by a simple extraction procedure without laborious column chromatography to afford both enantiomers of the alcohols. We then determined that the dicarboxylic acid monoesters were suitable alternative substrates for the preparation of optically active alcohols.

Full text of DOI:10.2174/157017812803521108

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Letters in Organic Chemistry, 2012, 9, 615-621
615
Enzyme-Mediated Enantioselective Hydrolysis of Dicarboxylic Acid
Monoesters
Noriyuki Kataoka, Masayuki Okudomi, Naoka Chihara and Kazutsugu Matsumoto*
Department of Chemistry and Life Science, Meisei University, 2-1-1 Hodokubo, Hino, Tokyo 191-8506, Japan
Received July 20, 2012: Revised August 17, 2012: Accepted August 23, 2012
Abstract: The enzyme-mediated highly enantioselective hydrolysis of dicarboxylic acid monoesters was investigated.
The racemic substrates, which were prepared by coupling of the corresponding alcohols with dicarboxylic anhydrides,
were enantioselectively hydrolyzed by lipase from Candida antarctica (Novozym 435) in a buffer at 30 °C. The products
were easily separated by a simple extraction procedure without laborious column chromatography to afford both enanti-
omers of the alcohols. We then determined that the dicarboxylic acid monoesters were suitable alternative substrates for
the preparation of optically active alcohols.
Keywords: Dicarboxylic acid monoesters, enzymatic hydrolysis, kinetic resolution, lipase, optically active alcohols.
INTRODUCTION
the optically active 3. We noted that the dicarboxylic acid
monoesters could be a good substrate for hydrolytic en-
zymes, because the remaining dicarboxylic acid monoesters
could be separated from the resulting alcohols by the same
process as in the case of the enzymatic esterification with
succinic anhydride as mentioned above. To the best of our
knowledge, there have been only a very few reports on the
enzyme-mediated enantioselective hydrolysis of dicarbox-
ylic acid monoesters (Fig. 2) [24, 25], and the E values (the
ratio of the specificity constants of enantiomers (R and S); E
value=(VR/KR)/(VS/KS)) [26] indicated that the enantioselec-
tivities of the reactions using an industrial glutaryl acylase
were low. However, the dicarboxylic acid monoesters are
well known for use as prodrugs, which are hydrolyzed by
hydrolases [27]. In addition, a free terminal carboxylate
group could cause an interaction between the substrates and
the enzyme active site residue through a hydrogen bonding
[28-30]. We now disclose the enzyme-mediated enantiose-
lective hydrolysis of dicarboxylic acid monoesters, and also
report the structure of the acyl moiety that affects both the
reactivity and enantioselectivity.
The enzyme-mediated kinetic resolution of racemic alco-
hols and esters is one of the attractive methods for the prepa-
ration of optically active compounds [1-11]. During the reac-
tion process, however, the products and the remaining sub-
strates must be separated by column chromatography, which
is the bottleneck to a sustainable production and an easy op-
eration. The use of succinic anhydride as an acylating agent
during enzymatic esterification could resolve this concern
[12-22]. Although the separation of the resulting dicarbox-
ylic monoesters from the remaining alcohols could be
achieved by a simple extraction process, the procedure is not
always satisfactory in terms of the enantioselectivity and the
necessity for high amount of enzymes.
In our previous study, we succeeded in the enanti-
oselective hydrolysis of monomethoxy poly(ethylene glycol)
(MPEG; av MW 5000)-supported carboxylates using lipase
from Candida antarctica (Novozym 435; CAL-B), and the
separation of the reaction products was achieved by simple
filtration because the MPEG-supported substrates were eas-
ily precipitated in Et2O [23]. At the beginning of the study,
we carried out the enzymatic hydrolysis of the MPEG-
supported diester (±)-1 (Fig. 1). Unfortunately, Novozym
435 catalyzed the hydrolysis of two ester bonds, and the sub-
strate was decomposed into several components. After the
detail analysis of the products, however, we found that the
MPEG ester part was first hydrolyzed to give the corre-
sponding succinic acid monoester (±)-2a, because hydrolases
generally preferred a primary ester. In addition, the resulting
racemate (±)-2a was then enantioselectively hydrolyzed to
RESULTS AND DISCUSSION
We selected 1-phenylethanol (3) as the representative al-
cohol, and the racemate (±)-3 was combined with succinic
anhydride using DMAP in CH2Cl2 to give the corresponding
substrate (±)-2a (Fig. 3). The other substrates were synthe-
sized by the same procedure.
At first, the enzymatic reaction using Novozym 435 of
the succinic acid monoester (±)-2a was carried out (Fig. 4).
In a typical experiment, 89 mg of (±)-2a (sub. concn. 10
mmol L-1) and 20 mg of Novozym 435 were added to 0.1
mol L-1 phosphate buffer (pH 6.5, 40 mL) in a recovery
flask, and the reaction was stirred for 24 h at 30 °C. The wa-
ter solubility of the substrate under suitable basic conditions
*Address corresponding to this author at Department of Chemistry and Life
Science, Meisei University, 2-1-1 Hodokubo, Hino, Tokyo 191-8506, Ja-
pan; Tel./Fax: +81 42 591 7360; E-mail: mkazu@chem.meisei-u.ac.jp
1875-6255/12 $58.00+.00
© 2012 Bentham Science Publishers

616 Letters in Organic Chemistry, 2012, Vol. 9, No. 9
Kataoka et al.
O
Me
O
Me
Novozym 435
HO
O
O
Ph
O
Ph
MPEG
O
O
(±)-1
(±)-2a
OH
Novozym 435
OH
*
Ph
Me
3*
Fig. (1). Enzymatic hydrolysis of a MPEG-supported diester.
Me
O
O
O
O
Me
OH
Glutaryl acylase
+
HO
X
O
Ph
HO
X
O
Ph
Ph
Me
H2O
(±)-2
(S)-2
(R)-3
CH2CH2
a: X =
b: X =
E value = 18.5
E value = 7.2
CH2CH2CH2
Fig. (2). Enzymatic hydrolysis of carboxylic acid monoesters with glutaryl acylase.
O
O
O
O
Me
OH
, DMAP
HO
O
Ph
Ph
Me
CH2Cl2
rt, 24 h
O
(±)-3
(±)-2a
Fig. (3). Preparation of the substrate (±)-2a.
O
Me
O
Me
OH
Novozym 435
HO
HO
+
O
Ph
O
Ph
pH 6.5 buffer
30 °C, 24 h
Ph
Me
O
O
(±)-2a
(S)-2a
Separation
(R)-3
conv. = 0.33
E value = 320
under basic condition
In organic phase
In aqueous phase
NaOH
OH
Ph
(S)-3
Me
Fig. (4). Enzymatic hydrolysis of the succinic acid monoester (±)-2a.
(ca. pH 11) enabled us to establish a facile separation of the
remaining 2a and the resulting alcohol 3. After acidification
of the reaction mixture with 2 mol L-1 HCl, the first extrac-
tion process was performed with Et2O. In this step, both the
substrate 2a and the alcohol 3 were extracted into the or-
ganic phase. In the following second extraction from the
organic phase with 1 mol L-1 Na2CO3 aq, the substrate 2a
was then selectively obtained in the basic aqueous phase.
The alcohol 3 successfully remained in the organic phase,
and was isolated after evaporation. On the other hand,
chemical hydrolysis of the substrate 2a by the addition of 2
mol L-1 NaOH aq to the aqueous phase gave the correspond-
ing 3, which was also extracted with Et2O. The yields of the
compounds were determined after passing through a small
amount of silica gel. The enantiomeric excesses (ee) were
evaluated by a chiral GLC analysis. As expected, the hy-
drolysis proceeded with a high enantioselectivity to afford
the optically active (S)-3 (derived from (S)-2a; 45%, 48%
ee) and (R)-3 (15%, 99% ee). Although the reactivity was
moderate (conv.=0.33; the conversion was calculated using
ees/(ees+eep); ees, ee of (S)-alcohol; eep, ee of (R)-alcohol),
the enantioselectivity (E value=320; the E value was calcu-
lated using ln[(1–conv.)(1–ees)]/ ln[(1–conv.)(1+ees)]) was
very high.

Enzyme-Mediated Enantioselective Hydrolysis of Dicarboxylic Acid
Me
Letters in Organic Chemistry, 2012, Vol. 9, No. 9
617
O
O
O
O
Me
OH
Novozym 435
+
HO
X
O
Ph
HO
X
O
Ph
pH 6.5 buffer
30 °C, 24 h
Ph
(R)-3
Me
(±)-2b-d
(S)-2b-d
NaOH
OH
a: X =
b: X =
CH2CH2
CH2CH2CH2
Me
c: X =
d: X =
CH2CHCH2
Me
Me
Ph
(S)-3
Me
CH2C CH2
Fig. (5). Enzymatic hydrolysis of the substrates bearing various acyl parts.
Table 1. Enantioselective hydrolysis of dicarboxylic acid monoesters 2b-d with Novozym 435.a
Entry
Substrate
(S)-3
(R)-3
Conv.b
E valuec
Yield (%)
Ee (%)d
Yield (%)
Ee (%)d
1
2e
3
(±)-2b
(±)-2b
(±)-2c
(±)-2d
34
42
73
91
96
16
38
41
10
>99
>99
97
0.48
0.49
0.14
–
>640
>790
77
4
No reaction
–
aThe reaction was performed using 10 mM of the substrate with Novozym 435 (20 mg) in 0.1 M phosphate buffer (pH 6.5) for 24 h at 30 °C.
bCalculated using ees/(ees+eep).
cCalculated using ln[(1–conv.)(1–ees)]/ ln[(1–conv.)(1+ees)].
dDetermined by GC analysis.
eThe reaction was performed on a gram scale of the substrate.
We next investigated changing the acyl group of the sub-
strate in order to increase the conversion rate, because we
speculated that a suitable acyl group could increase the affin-
ity to the active site of the enzyme (Fig. 5). The results are
summarized in Table 1. Surprisingly, changing the acyl
group significantly affected not only the conversion, but also
the enantioselectivity. The glutaric acid monoester (±)-2b
was smoothly hydrolyzed, and the conversion was up to 0.48
(entry 1). In addition, an excellent enantioselectivity was
observed (E value>640) to afford both almost optically pure
enantiomers. This reaction was also useful as a preparative-
scale operation (entry 2, >1 g of (±)-2b). Finally, both the
reactivity and enantioselectivity in the case of the dicarbox-
ylic acid monoester 2b were eventually comparable to those
for the enzymatic hydrolysis of the racemic 1-phenylethyl
acetate ((±)-4), which was a typical substrate, under the same
reaction conditions (conv.=0.50, E value=920). On the other
hand, the reactivity of the 3-methylglutaric acid monoester
(±)-2c significantly decreased (conv.=0.14) under the same
reaction conditions, and the enantioselectivity (E value=77)
was also lower than that for 2a and 2b (entry 3). As ex-
pected, the hydrolysis reaction of the compounds 2d bearing
a bulky substituent (3,3-dimethyl group) did not proceed at
all (entry 4). Fiaud et al. reported the esterifications of (±)-3
with dicarboxylic anhydride using the same enzyme in Et2O
for 24 h, and the enantioselectivities were almost same as
those obtained by our hydrolysis version [22]. However, in
this study, the enzymatic esterification of 3 with glutaric
anhydride (conv.=0.23) was slower than that for the reaction
with succinic anhydride (conv.=0.50). Interestingly, the re-
sults were apparently contrary to the effect of the acyl moi-
ety in our cases.
In order to apply the concept of this reaction for the ki-
netic resolution of other secondary alcohols, we next exam-
ined the enzymatic hydrolysis of several succinic and glu-
taric acid monoesters (Fig. 6), and these results are shown in
Table 2. In all cases, the reactions were performed for a
longer reaction time (48 h) using a higher amount of the en-
zyme (40 mg), because the reactivities were lower than that
in the case of the substrate 2b. While the hydrolysis of the
succinic acid monoester (±)-5a bearing an ethyl group as the
R2 substituent (R1=Ph, R2=Et) with a moderate enantioselec-
tivity was very slow (entry 1; conv.=0.08, E value=72),
changing the acyl moiety to the glutaric acid monoester ((±)-
5b) drastically improved both the reactivity and enantiose-
lectivity (entry 2; conv.=0.35, E value=340). In the case of
(±)-6b bearing a phenylmethyl group (R1=CH2Ph, R2=Me),
the enzyme also catalyzed the hydrolysis of the compound
with a higher enantioselectivity (conv.=0.46, E value=520)
than that of (±)-6a (conv.=0.38, E value=180) to afford the
highly optically active compounds (entries 3 and 4). On the
other hand, the reactions of the substrates bearing a naphthyl
group as the R1 substituent also proceeded. Although the
enantioselectivities of the esters ((±)-8a and 8b, R1=1-
naphthyl, R2=Et) of 1-(1-naphthyl)ethanol (13) were low
(entries 7 and 8; E value = 16 and 18, respectively),
both esters ((±)-7a and 7b, R1=2-naphthyl, R2=Et) of 1-(2-

618 Letters in Organic Chemistry, 2012, Vol. 9, No. 9
Kataoka et al.
R2
O
O
R2
O
O
OH
Novozym 435
+
R1
HO
X
O
R1
HO
X
O
R1
R2
pH 6.5 buffer
30 °C, 48 h
(±)-5-9
(S)-5-9
NaOH
OH
(R)-10-14
5, 10: R1 = Ph, R2 = Et
6, 11: R1 = CH2Ph, R2 = Me
R1
R2
7, 12: R1 =
, R2 = Me
(S)-10-14
8, 13: R1 =
, R2 = Me
a: X =
b: X =
CH2CH2
9, 14: R1 = (CH2)2OBn, R2 = Me
CH2CH2CH2
Fig. (6). Enzymatic hydrolysis of the various carboxylic acid monoesters.
Table 2. Enantioselective hydrolysis of dicarboxylic monoesters 5-9 with Novozym 435. a
(S)-alcoholb
Yield (%)
(R)-alcoholb
Yield (%)
Entry
Substrate
Conv.c
E valued
Ee (%)
Ee (%)
1
2
(±)-5a
(±)-5b
(±)-6a
(±)-6b
(±)-7a
(±)-7b
(±)-8a
(±)-8b
(±)-9a
(±)-9b
64
54
48
38
50
35
72
75
48
36
9
10
17
22
39
49
35
10
11
44
45
97
99
98
99
99
98
87
88
99
97
0.08
0.35
0.38
0.46
0.49
0.50
0.09
0.12
0.50
0.51
72
55
59
83
97
99
9
340
180
520
840
530
16
3
4
5
6
7
8
12
98
99
18
9
920
350
10
aThe reactions were performed using 10 m mol L-1 of the substrate with Novozym 435 (40 mg) in 0.1 mol L-1 phosphate buffer (pH 6.5) for 48 h at 30 °C.
bThe absolute configurations were determined by comparing the specific rotation signs with those reported.
cCalculated using ees/(ees+eep).
dCalculated using ln[(1–conv.)(1–ees)]/ ln[(1–conv.)(1+ees)].
OAc
Me
OAc
Me
OH
Novozym 435
+
pH 6.5 buffer
30 °C, 24 h
BnO
BnO
BnO
Me
(±)-15
(S)-15
45%, 94% ee
(R)-14
52%, 86% ee
conv. = 0.52
E value = 47
Fig. (7). Enzymatic hydrolysis of the acetate (±)-15 with Novozym 435.
naphtyl)ethanol (12) were hydrolyzed with excellent enanti-
oselectivities (entries 5 and 6; E value=840 and 530, respec-
tively). It is noteworthy that the kinetic resolutions of (±)-9a
value=920 and 350, respectively). We also examined the
enzymatic hydrolysis of the acetate (±)-15 for 24 h at 30 °C
(Fig. 7). Although the enzymatic hydrolysis smoothly pro-
ceeded, the enantioselectivity was only moderate
(conv.=0.52, E value=47). These results indicated that the
structure of the acyl moieties of 9 apparently affects the in-
and 9b, which contain
a
benzyloxyethyl group
(R1=CH2CH2OBn, R2=Me), were smoothly accomplished to
afford almost optically pure compounds (entries 9 and 10; E

Enzyme-Mediated Enantioselective Hydrolysis of Dicarboxylic Acid
Letters in Organic Chemistry, 2012, Vol. 9, No. 9
619
13C NMR (125 MHz, CDCl3) ꢀ 22.1, 28.9, 29.1, 72.9, 126.0,
127.9, 128.5, 141.3, 171.4, 178.2; HRMS m/z (ESI)
245.0788 (calcd for C12H14O4Na: 245.0790, M+Na+).
teraction between the substrate and the active site of the en-
zyme.
Except for the substrate 13 bearing a 1-naphthyl group,
all the reactions gave enough high E values (over 180), re-
gardless of the size of the R1 substituent. We then supposed
that many aromatic and aliphatic R1 substituents, which had
a suitable size, could be acceptable for the substrates. We
also speculated that a 1-naphthyl group would be too bulky
for the interaction between the substrate and the enzyme due
to the position of the naphthyl group. On the other hand, the
conversions for the substrates with an ethyl group as the R2
substituent (5a and 5b) were lower than those for the sub-
strates with a methyl group, and we supposed that the size of
the R2 substituent should be restricted. In order to improve
the conversion for the practical synthesis, a much longer
reaction time and a much higher amount of the enzyme
could be required.
The other substrates were synthesized by the same pro-
cedure.
Typical Experimental Procedure for Enantioselective
Hydrolysis of Dicarboxylic Monoesters (1-phenylethanol
(3))
To a recovery flask containing 88.9 mg of (±)-2a (sub.
conc., 10 mmol L-1) were added 40 mL of 0.1 mol L-1 phos-
phate buffer (pH6.5). To the mixture was added 20 mg of
Novozym 435 (2.0 U/mg, using tributyrin at pH 8.0 and 40
°C), and the solution was stirred for 24 h at 30 °C. After ad-
dition of 2 mol L-1 HCl to the mixture, the products were
extracted with Et2O (x4), and dried over Na2SO4. After the
organic phase was evaporated in order to reduce the volume,
(S)-2a was selectively extracted with 1 mol L-1 Na2CO3 aq
(x4). Evaporation of the remaining organic phase afforded
(R)-3. On the other hand, to the water phase was added 2
mol L-1 NaOH aq (5 mL), and the mixture was stirred for 2 h
at room temperature. The products were extracted with Et2O
(x4), and dried over Na2SO4. Evaporation of the organic
phase afforded (S)-3. In order to determine the exact isolated
yields and ees of the products, both enantiomers were puri-
fied by column chromatography on a small amount of silica
CONCLUSION
In conclusion, we succeeded in the enzyme-mediated
enantioselective hydrolysis of dicarboxylic acid monoesters,
and obtained several enantiomers of 3, 10, 11, 12, 13, and
14. We also have disclosed that the reactivity and eantiose-
lectivity can be controlled using a suitable acyl group of the
substrates. In our method, the separation of the reaction
products was achieved by a simple extraction procedure.
Further investigations into the reactions are now in progress.
gel (hexane/Et2O = 2/1) to give (S)-3 (22.0 mg, 45%, 48%
28
ee) and (R)-3 (7.5 mg, 15%, 99% ee), respectively. [ꢀ]D
=
27
–43.0 (c 1.13, MeOH) (96% ee, (S)-form), [ꢀ]D = +41.0 (c
1.30, MeOH) (>99% ee, (R)-form); lit. [ꢀ]D = +45 (c 5.15,
20
MeOH) ((R)-form) [31]. GC conditions: column, CP-
Cyclodextrin-B-236-M19 (Chrompack), 0.25 mm x 50 m;
injection, 140 °C; detection, 140 °C; oven, 120 °C; carrier
gas, He; head pressure, 2.4 kg/cm2; retention time, 14.6 (R)
and 15.2 (S) min.
EXPERIMENTAL
General
1H (500 MHz or 300 MHz) and 13C (125 MHz or 75
MHz) NMR spectra were measured on a JEOL ꢀ-500 or a
JEOL JNM AL-300 with tetramethylsilane (TMS) as the
internal standard. IR spectra were recorded with Shimadzu
IR Prestige-21 spectrometers. Mass spectra were obtained
with a JEOL EI/FAB mate BU25 Instrument by the EI
method or a JEOL JMS-T100 by the ESI-TOF method. Op-
tical rotations were measured with a Jasco DIP-1000 po-
larimeter. HPLC data were obtained on Shimadzu LC-
10ADVP, SPD-10AVP, and sic 480II data station (System
Instruments Co., Ltd.).
The reactions of the other substrates were carried out by
the same procedure. The results were shown in the text. All
the spectral data (1H and 13C NMR, IR, and MS) were in full
agreement with those of the racemates, commercial sources,
or those reported.
1-phenyl-1-propanol (10)
25
[ꢀ]D = +42.8 (c 0.93, CHCl3) (99% ee, (R)-form); lit.
20
[ꢀ]D = –47.0 (c 1.00, CHCl3) ((S)-form) [32]. GC condi-
tions: column, CP-Cyclodextrin-B-236-M19 (Chrompack),
0.25 mm x 50 m; injection, 140 °C; detection, 140 °C; oven,
120 °C; carrier gas, He; head pressure, 2.4 kg/cm2; retention
time, 22.6 (R) and 23.4 (S) min.
Preparation
of
the
Substrate,
(±)-3-((1-
phenylethoxy)carbonyl)Propanoic Acid (2a)
Under an argon atmosphere, succinic anhydride (490 mg,
4.89 mmol) and DMAP (599 mg, 4.901 mmol) were added
to a solution of 1-phenyletanol ((±)-3, 299.1 mg, 2.45 mmol)
in CH2Cl2 (10 mL), and the solution was stirred for 2 h at
room temperature. After the mixture was washed with 2 mol
L-1 HCl, the products were extracted with CH2Cl2 (x4), and
dried over Na2SO4. After evaporation in vacuo, the residue
was purified by column chromatography on silica gel (hex-
ane/AcOEt = 3/1) to give the (±)-2a as a colorless oil in 97%
yield (528 mg); IR (neat) 2982, 2934, 1736, 1713, 1495,
1450, 1375, 1285, 1207, 1169, 1063, 959, 762, 700 cm-1; 1H
NMR (500 MHz, CDCl3) ꢀ 1.53 (d, J = 6.5 Hz, 3H), 2.56-
2.74 (m, 4H), 5.89 (q, J = 6.5 Hz, 1H), 7.21-7.41 (m, 5H);
1-phenyl-2-propanol (11)
28
[ꢀ]D = –35.4 (c 0.75, CHCl3) (99% ee, (R)-form); lit.
20
[ꢀ]D = –37.6 (c 5.00, CHCl3) ((R)-form) [33]. GC condi-
tions: column, CP-Cyclodextrin-B-236-M19 (Chrompack),
0.25 mm x 50 m; injection, 130 °C; detection, 130 °C; oven,
110 °C; carrier gas, He; head pressure, 2.4 kg/cm2; retention
time, 27.1 (R) and 27.5 (S) min.
1-(2-naphthyl)ethanol (12)
Mp 68.2-68.8 °C (recrystallized from Et2O-hexane); lit.
28
68-70 °C [34]. [ꢀ]D = +38.3 (c 1.01, MeOH) (98% ee, (R)-
form); lit. [ꢀ]D = +34.6 (c 1.20, MeOH) ((R)-form) [35]. GC

620 Letters in Organic Chemistry, 2012, Vol. 9, No. 9
Kataoka et al.
[12]
[13]
Terao, Y.; Tsuji, K.; Murata, M.; Achiwa, K.; Nishio, T.; Wata-
nabe, N.; Seto, K. Facile process for enzymatic resolution of race-
mic alcohols. Chem. Pharm. Bull., 1989, 37, 1653-1655.
Gutman, A.L.; Brenner, D.; Boltanski, A. Convenient practical
resolution of racemic alkyl-aryl alcohols via enzymatic acylation
with succinic anhydride in organic solvents. Tetrahedron: Asymme-
try, 1993, 4, 839-844.
conditions: column, CP-Cyclodextrin-B-236-M19 (Chrom-
pack), 0.25 mm x 50 m; injection, 180 °C; detection, 180 °C;
oven, 160 °C; carrier gas, He; head pressure, 2.4 kg/cm2;
retention time, 41.9 (R) and 42.9 (S) min.
1-(1-naphthyl)ethanol (13)
[14]
[15]
[16]
[17]
[18]
[19]
Patel, R.; Banerjee, A.; Nanduri, V.; Goswami, A.; Comezoglu, F.
T. Enzymatic resolution of racemic secondary alcohols by lipase B
from Candida antarctica. J. Amer. Oil Chem. Soc., 2000, 77, 1015-
1019.
Bouzemi, N.; Debbeche, H.; Aribi-Zouioueche, L.; Fiaud, J. –C.
On the use of succinic anhydride as acylating agent for practical
resolution of aryl-alkyl alcohols through lipase-catalyzed acylation.
Tetrahedron Lett., 2004, 45, 627-630.
Rasalkar, M.S.; Potdar, M.K.; Salunlhe, M.M. Pseudomonas ca-
pacia lipase-catalyzed resolution of racemic alcohols in ionic liquid
using succinic anhydride: role of triethylamine in enhancement of
catalytic activity. J. Mol. Catl. B: Enzymatic, 2004, 27, 267-270.
Miyazawa, T.; Kaito, E.; Yukawa, T.; Murashima, T.; Yamada, T.
Enzymatic resolution of 2-aryloxy-1-propanols via lipase-catalyzed
enantioselective acylation using anhydride as acyl donors. J. Mol.
Catl. B: Enzymatic, 2005, 37, 63-67.
Zada, A.; Dunkelblum, E. A convenient resolution of racemic
lavandulol through lipase-catalyzed acylation with succinic anhy-
dride: simple preparation of enantiomerically pure (R)-lavandulol.
Tetrahedron Asymmetry, 2006, 17, 230-233.
Wang, Y.; Wang, R.; Li, Q.; Zhang, Z.; Feng, Y. Kinetic resolution
of rac-alkyl alcohols via lipase-catalyzed enantioselective acylation
using succinic anhydride as acylating agent. J. Mol. Catl. B: Enzy-
matic, 2009, 56, 142-145.
Maywald, M.; Pfalts, A. Chromatography-free enzymatic kinetic
resolution of secondary alcohols. Synthesis, 2009, 3654-3660.
Lourenco, N.M. T.; Monteiro, C.M.; Afonso, C. A. M. Ionic acy-
lating agents for the enzymatic resolution of sec-alcohols in ionic
liquids. Euro. J. Org. Chem., 2010, 6938-6943.
Debbeche, H.; Toffano, M.; Fiaud, J.–C.; Arbi-Zouioueche, L.
Multi-substrate screening for lipase-catalyzed resolution of arylal-
kylethanols with succinic anhydride as acylating agent. J. Mol. Cat.
B: Enzymatic, 2010, 66, 319-324.
Okudomi, M.; Ageishi, K.; Yamada, T.; Chihara, N.; Nakagawa,
T.; Mizuochi, K.; Matsumoto, K. Enzyme-mediated enantioselec-
tive hydrolysis of soluble polymer-supported carboxylates. Tetra-
hedron, 2010, 66, 8060-8067.
Raimondi, S.; Monti, D.; Forti, L.; Riva, S. Kinetic resolutions of
racemic amines and alcohols catalyzed by an industrial glutaryl-7-
aminocephalosporanic acid acylase with unexpected broad sub-
strate specificity. Tetrahedron Asymmetry, 2003, 14, 1091-1094.
Adani, S.; Raimondi, S.; Forti, L.; Monti, D.; Riva, S. Enantiose-
lective esterase activity of an industrial glutaryl acylase. Tetrahe-
dron Asymmetry, 2005, 16, 2509-2513.
Chen, C.–S.; Fujimoto, Y.; Grdaukas, G.; Sih, C. J. Quantitative
analyses of biochemical kinetic resolutions of enantiomers. J. Am.
Chem. Soc., 1982, 104, 7294-7299.
Stella, V.J.; Borchardt, R.T.; Hageman, M.J.; Oliyai, R.; Maag, H.
Tilley, J.W. Prodrugs: Challenges and Rewards (Biotechnology:
Pharmaceutical Aspects). Springer: New York, 2007.
Sobrado, P.; Fitzpatrick, P.F. Identification of Tyr413 as an Active
Site Residue in the Flavoprotein Tryptophan 2-monooxygenase and
analysis of its contribution to catalysis. Biochemistry 2003, 42,
13833-13838.
Okada, T.; Suzuki, H.; Wada, K.; Kumagai, H.; Fukuyama, K.
Crystal structures of g-glutamyltranspeptidase from E. coli, a key
enzyme in glutathione metabolism, and its intermediate. Proc. Natl.
Acad. Sci. USA, 2006, 103, 6471-6476.
Yamazawa, R.; Nakajima, Y.; Mushiake, K.; Yoshimoto, T.; Ito,
K. Crystal structure of serine dehydrogenase from Escherichia coli:
Important role of the C-terminal region for closed-complex forma-
tion. J. Biochem., 2011, 149, 701-712.
Laumen, K.; Schneider, M.P. A highly selective ester hydrolase
from Pseudomonas sp. for the enzymatic preparation of enanti-
omerically pure secondary alcohols; chiral auxiliaries in organic
synthesis. J. Chem. Soc., Chem. Commun., 1988, 598-600.
Nakamura, K.; Fujii, M.; Ida, Y. Stereoinversion of arylethanols by
Geotrichum candidum. Tetrahedron: Asymmetry, 2001, 12, 3147-
3153.
Mp 62.0-62.3 °C (recrystallized from CH2Cl2-hexane);
27
lit. 64 °C [36]. [ꢀ]D = +55.9 (c 0.64, MeOH) (88% ee, (R)-
25
form); lit. [ꢀ]D = +45.0 (c 2.00, MeOH) ((R)-form)[37].
GC conditions: column, CP-Cyclodextrin-B-236-M19
(Chrompack), 0.25 mm x 50 m; injection, 180 °C; detection,
180 °C; oven, 160 °C; carrier gas, He; head pressure, 2.4
kg/cm2; retention time, 43.0 (R) and 44.3 (S) min.
4-benzyloxy-2-butanol (14)
29
[ꢀ]D = –14.9 (c 1.08, MeOH) (99% ee, (R)-form); lit.
[ꢀ]D27 = +19.0 (c 0.95, MeOH) ((S)-form)[38]. HPLC condi-
tions: column, CHIRALCEL OD-H (Daicel Chemical Indus-
tries, Ltd.); eluent, hexane/2-propanol = 90/10; flow rate, 0.5
mL/min; 254 nm; temperature, 25 °C; retention time, 12.8
(S) and 14.0 (R) min.
CONFLICT OF INTEREST
The author(s) confirm that this article content has no con-
flicts of interest.
[20]
[21]
ACKNOWLEDGEMENTS
We thank Collaborative Research Center of Meisei Uni-
versity for NMR analysis.
[22]
[23]
[24]
SUPPLEMENTARY MATERIAL
Supplementary material (GC and HPLC charts) is avail-
able on the publishers website along with the published arti-
cle.
REFERENCES
[25]
[26]
[27]
[28]
[1]
[2]
[3]
[4]
Bornscheuer, U.T.; Kazlauskas, R.J. Hydrolases in Organic Syn-
thesis, Wiley-VCH: Weinheim, 1999.
Faber, K. Biotransformations in Organic Chemistry: A Textbook,
5th ed.; Springer Verlag: Berlin-Heidelberg-New York, 2004.
Ghanem, A.; Aboul-Enein, H.Y. Application of lipases in kinetic
resolution of racemates. Chirality, 2005, 17, 1-15.
Garcia-Uradiales, E.; Alfonso, I.; Gotor, V. Enantioselective En-
zymatic Desymmetrizations in Organic Synthesis. Chem. Rev.,
2005, 105, 313-354.
[5]
[6]
Bornscheuer, U.T. Trends and Challenge in Enzyme Technology,
Springer: Berlin/Heidelberg, 2005.
Fogassy, E.; Nógrádi, E.; Kozma, D.; Egri, G.; Pálovics, E.; Kiss,
V. Optical resolution methods. Org. Biomol. Chem., 2006, 4, 3011-
3030.
Gadler, P.; Faber, K. New enzymes for biotransformations: micro-
bial alkyl sulfatases displaying stereo- and enantioselectivity.
Trends Biotechnol., 2007, 25, 83-88.
Ghanem, A. Trends in lipase-catalyzed asymmetric access to enan-
tiomerically pure/enriched compounds. Tetrahedron, 2007, 63,
1721-1754.
Gotor, V.; Alfonso, I.; Garcia-Urdiales, E. Asymmetric Organic
Synthesis with Enzymes, Wiley-VCH: Weinheim, 2008.
Whittall, J.; Sutton, P.W. Practical Methods for Biocatalysis and
Biotransformations, Wiley: West Sussex, 2010.
[29]
[30]
[31]
[32]
[7]
[8]
[9]
[10]
[11]
Drauz, K.; Gröger, H.; May, O. Enzyme Catalysis in Organic Syn-
thesis, 3rd ed.; Wiley-VCH: Weinheim, 2012.

Enzyme-Mediated Enantioselective Hydrolysis of Dicarboxylic Acid
Letters in Organic Chemistry, 2012, Vol. 9, No. 9
621
[33]
Masuda, S.; Nakajima, T.; Suga, S. Retentive Friedel-Crafts alkyla-
tion of benzene with optically active 2-chloro-1-phenylpropane and
1-chloro-2-phenylpropane. Bull. Chem. Soc. Jpn., 1983, 56, 1089-
1094.
Liu, P.N.; Deng, J.G.; Tu, Y.Q.; Wang, S.H. Highly efficient and
recyclable heterogeneous asymmetric transfer hydrogenation of ke-
tones in water. Chem. Commun., 2004, 2070-2071.
Ziffer, H.; Kawai, K.; Kasai, M.; Imuta, M.; Froussios, C. Microbi-
ally mediated enantioselective ester hydrolyses utilizing Rhizopus
nigricans. A new method of assigning the absolute stereochemistry
of acyclic 1-arylalkanols. J. Org. Chem., 1983, 48, 3017-3021.
[36]
[37]
Rahaim, R. J.; Maleczka, R. E., Jr. CꢀO Hydrogenolysis catalyzed
by Pd-PMHS nanoparticles in the company of chloroarenes. Org.
Lett., 2011, 13, 584-587.
Kanth, J.V.B.; Periasamy, M. Asymmetric reduction of prochiral
aromatic ketones by Borane–Amine complexes in the presence of a
chiral amine–BF3 catalyst. J. Chem. Soc., Chem. Commun., 1990,
1145-1147.
Nogawa, M.; Shimojo, M.; Matsumoto, K.; Okudomi, M.; Nemoto,
Y.; Ohta, H. Kinetic resolution of poly(ethylene glycol)-supported
carbonates by enzymatic hydrolysis. Tetrahedron, 2006, 62, 7300-
7306.
[34]
[35]
[38]