Microbial asymmetric reduction of α-hydroxyketones in the anti-Prelog selectivity

Article abstract of DOI:10.1016/S0957-4166(01)00448-7

Yamadazyma farinosa IFO 10896 was found to reduce α-hydroxyketones bearing a phenyl ring to give optically active diols with anti-Prelog selectivity. The distance between the carbonyl group and the phenyl ring was shown to have an interesting effect on the reactivity and selectivity of the enzyme system.

Full text of DOI:10.1016/S0957-4166(01)00448-7

TETRAHEDRON:
ASYMMETRY
Pergamon
Tetrahedron: Asymmetry 12 (2001) 2543–2549
Microbial asymmetric reduction of a-hydroxyketones in the
anti-Prelog selectivity
Toshikuni Tsujigami, Takeshi Sugai and Hiromichi Ohta*
Department of Chemistry, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan
Received 29 August 2001; accepted 11 October 2001
Abstract—Yamadazyma farinosa IFO 10896 was found to reduce a-hydroxyketones bearing a phenyl ring to give optically active
diols with anti-Prelog selectivity. The distance between the carbonyl group and the phenyl ring was shown to have an interesting
effect on the reactivity and selectivity of the enzyme system. © 2001 Elsevier Science Ltd. All rights reserved.
1. Introduction
In this way, it is desirable to find many kinds of
microorganisms which have their own characteristics in
their substrate specificity and selectivity to fill the vari-
ety of demands from asymmetric synthesis. Although
baker’s yeast has established to have plural reducing
enzymes,6,7 it generally gives the (S)-alcohol on reduc-
tion of ketones,1 known as Prelog’s rule.8
The most commonly used biocatalyst in asymmetric
reduction is baker’s yeast. This microorganism is com-
mercially available, easy to handle, and able to be used
in the asymmetric reduction of wide range of carbonyl
functionalities and carbonꢀcarbon double bonds.1
Recently, the scope of biotransformation seems to be
expanding increasingly wider, as the whole cell biocata-
lysts have been shown to be used in solvents other than
aqueous medium. For example Nakamura et al.
showed that baker’s yeast can be used in organic
solvents2 and resting cells of Geotricum were demon-
strated to work well in supercritical carbon dioxide by
Matuda et al.3 Moreover, microbial reduction can be
performed on a large scale so that it can be applied in
an industrial process by utilizing an engineered
microorganism4 and designing the reaction conditions.5
We have screened for a microorganism which reduces
carbonyl compounds in an anti-Prelog manner and
found that Yamadazyma farinosa is a useful strain
which is registered to the Institute for Fermentation,
Osaka with the registry number IFO 10896. This bio-
catalyst promotes the reduction of various ketones and
exhibits enantioselectivity to those which have a p-elec-
tron system or heteroatom moiety. Typical products
from the corresponding ketones are shown in Fig. 1.9
HO
H
HO
H
HO H
CH3
PhS
CH3
PhSO2
O
CH3
1
2
3
O
O
CH3
HO HO
H
HO
H
H
O
CH3
H3C
HO
CH3
H3C
CH3
H
4
5
6
7
O
S
CH3O
HO
HO
H
HO
H
H
PhS
CF3
Ph
CF3
H3CO
CH3
8
9
10
Figure 1. Optically active alcohols obtained via the Y. farinosa-mediated reduction of the corresponding ketones. See Refs. 9a for
1, 9b and 9f for 2 and 3, 9c for 4 and 5, 9d for 6 and 7, 9f for 8 and 9, and 9g for 10.
* Corresponding author. Tel./fax: +81-(0)45-566-1703; e-mail: hohta@chem.keio.ac.jp
0957-4166/01/$ - see front matter © 2001 Elsevier Science Ltd. All rights reserved.
PII: S0957-4166(01)00448-7

2544
T. Tsujigami et al. / Tetrahedron: Asymmetry 12 (2001) 2543–2549
Table 1. Asymmetric reduction of a-hydroxyacetophenone derivatives
O
HO H
OH
OH
Y. farinosa IFO 10896
(1)
phosphate buffer
R
R
(S)-12
a R: H, b R: CH3O, c R: Cl
11
Entry
R
Atmosphere
Conc. (% w/v)
Time (h)
Yield (%)
Config.
E.e. (%)
1
2
3
4
H
MeO
Cl
Ar
Ar
Ar
Air
0.5
0.5
0.3
0.5
24
48
48
24
91
95
85
86
S
S
S
S
\99
\99
\99
\99
H
As seen from these compounds, the substrates so far
subjected to the reduction system of Y. farinosa were
limited to methyl or trifluoromethyl ketones. Aiming to
widen the applicability of this unique biocatalyst, we
tried the reaction using substrates which are expected to
give chiral products for which CꢀC bond elongation on
both sides of the stereogenic center is possible.
formed ketol 4a via tautomerism prior to the second-
step reduction (Eq. (2)). If the same type of equilibrium
is present for hydroxyacetophenone derivatives 11, they
might possibly isomerize to hydroxy aldehydes 14 as
shown in Eq. (3), and if it were the real substrates of
microbial reduction, then the e.e. of the final product
would not be necessarily high. The sharp enantioselec-
tivity actually observed indicates that biotransforma-
tion of hydroxyacetophenone 11 is the direct reduction
of the ketone moiety of the added substrates. Another
interesting feature of the present reduction is that the
presence of air has no effect on the enantioselectivity in
contrast to the reduction of other compounds so far
investigated, where the presence of air lowered the e.e.
of the products.
2. Results and discussion
Y. farinosa was grown on a nutrient medium and
harvested by centrifugation. The reduction was per-
formed using resting cells in a phosphate buffer under
an atmosphere of argon. As shown in Table 1 (entries
1–3), a-hydroxyacetophenone and p-substituted deriva-
tives gave the corresponding diol in high yields and
high e.e., regardless to the electronic properties of the
substituents (Eq. (1)). Redley et al. have reported that
baker’s yeast reduces 11 to give (R)-styrene glycol 12.10
Thus, the enantioselectivity of the present biotransfor-
mation is opposite to that obtained by using intact cells
of baker’s yeast. Some years ago, we demonstrated that
this yeast reduces 1,3- and 1,4-diones to the corre-
sponding (R,R)-diols with high e.e. However, in con-
trast to these, in the case of butan-2,3-diones 3, the
yeast gave the meso-isomer 5 as the major product.9c
The mechanism of formation of meso-isomer was sup-
posed to be due to the racemization of intermediary
Although it is possible to transform a phenyl ring to a
carbonyl group, which can be used for CꢀC bond
formation, it would be more convenient if the substrate
has some protected functional groups, which will be
converted more easily to the active forms. Thus, we
tried the reaction with mono-protected dihydroxyacet-
ones 15a and 15b (Table 2 and Eq. (4)). In both cases,
the microbial reduction proceeded, although the rates
of the reaction was very slow for benzoyl derivatives
15b. The spatial arrangement of the ligands around the
stereogenic center of mono-benzylated triol 16a was the
same as that in the case of reduction of hydroxyacet-
O
H
(R)
OH
Y. farinosa
H3C
H3C
(S) CH3
CH3
HO
H
O
3
meso-5
(2)
(3)
OH
CH3
HO
H
H
OH
H3C
H3C
H3C
CH3
CH3
(S)
(R)
O
OH
4b
O
4a
4c
O
OH
OH
O
OH
OH
Ph
Ph
Ph
11a
13
14

T. Tsujigami et al. / Tetrahedron: Asymmetry 12 (2001) 2543–2549
Table 2. Asymmetric reduction of mono-protected dihydroxyacetone
2545
HO H
O
OH
OH
Y. farinosa IFO 10896
O
(R)-16a
(4)
O
OH
phosphate buffer
X
15
H OH
a R: CH2, b R: C=O
O
O
(S)-16b
Entry
X
Conc. (% w/v)
Time (h)
Yield (%)
Config.
E.e. (%)
1
2
CH2
CO
0.5
0.5
48
168
98
65
R
S
73
80
ophenone, although the absolute configuration was
opposite because of the priority rule. However, the
mode of reaction of benzoyl derivative 15b was entirely
different from those mentioned so far. The configura-
tion of the resulting mono-protected glycerol was
shown to be (S) on the basis of the sign of the optical
rotation and the order of elution in HPLC. The struc-
tural difference between the substrate 15a and 15b is
that 15b has a carbonyl group instead of a methylene of
15a. The difference of the polarity the facility in the
formation of hydrogen bonding, and other electronic
and steric effects are supposed to be enough for the
inversion of the binding mode. In fact, the free energy
required to reverse the two binding modes from the
ratio of 95/5 to 5/95 can be calculated to be only 3.5
kcal/mol at room temperature. The selectivity of the
reaction is lower compared to the reaction of 11. We
supposed that this may be attributed to the long dis-
tance between the carbonyl carbon and the phenyl ring.
In the case of compound 11, the aromatic ring is
directly attached to the carbonyl carbon, whilst in the
case of dihydroxyacetone derivative 15, there are three
atoms (CꢀOꢀC) between the carbonyl group and the
phenyl ring.
17a and 17b (Eq. (5)), the e.e. of the products were
better than those of 16, as expected. Thus, it can be
concluded that the distance between the phenyl ring
and the reaction center is important for the binding
mode of the substrates to the active site of the enzyme.
3. Conclusion
Various types of a-hydroxyketones bearing a phenyl
ring on the other side of the carbonyl group were
reduced with good to high enantioselectivities by the
aid of the enzyme system of Y. farinosa. The enantiose-
lectivity was anti-Prelog in most cases as expected. The
numbers of atoms between the carbonyl group and the
phenyl ring had a clear effect on the enantioselectivity
of the reaction, the nearer the phenyl ring to the
carbonyl group, the higher the e.e. of the products.
4. Experimental
4.1. General
All melting points were uncorrected. Optical rotation
values were recorded on a Jasco DIP 360 polarimeter.
IR spectra were measured as films for oils and as KBr
discs for solids with a Jasco FT/IR-410 instrument. H
NMR spectra were recorded at 270 MHz on a JEOL
JNM-EX 270 unless otherwise stated. Column chro-
To examine the effect of carbon numbers, two kinds of
ketones which have two atoms between the phenyl ring
and the reaction center were designed and subjected to
the reaction. The results are summarized in Table 3.
The reaction proceeded smoothly. For both compounds
1
Table 3. Asymmetric reduction of a-hydroxyketones with a two-atom spacer between the carbonyl group and the phenyl
ring
O
HO H
X
OH
X
OH
Y. farinosa IFO 10896
(5)
phosphate buffer
(R)- or (S)-18
a R: CH2, b R: O
17
Entry
X
Conc. (% w/v)
Time (h)
Yield (%)
Config.
E.e. (%)
1
2
CH2
O
0.5
0.5
48
48
94
63
S
R
94
88

2546
T. Tsujigami et al. / Tetrahedron: Asymmetry 12 (2001) 2543–2549
matography was carried out with Katayama 60 K070-
WH 70–230 mesh silica gel.
3H), 4.80 (s, 2H), 6.96 (d, J=9.07, 2H), 7.87 (d, J=
9.07, 2H).
4.2. Synthesis of substrates
4.2.2. p-Chloro-a-hydroxyacetophenone 11c. To a stirred
solution of m-CPBA (1.27 g, 7.36 mmol) in chloroform
(12 mL), p-chlorostyrene (1.03 g, 7.43 mmol) was
added in small portions over a period of 5–10 min at
0°C and the mixture was stirred overnight at rt. Then
1% (w/v) aqueous solution of sulfuric acid (36 mL) was
added and the mixture was stirred at rt for 1 h. 10%
NaOH (30 mL) was added and the aqueous solution
was extracted with ethyl acetate (5×10 mL). The com-
bined organic extract was washed with brine and dried
over anhydrous Na2SO4, and concentrated in vacuo.
The residue was purified by silica-gel column chro-
matography. Elution with hexane/ethyl acetate (1/1)
afforded 1-(p-chlorophenyl)-1,2-ethanediol (304 mg,
23%).
4.2.1. p-Methoxy-a-hydroxyacetophenone 11b (general
procedure for the selective oxidation of secondary
hydroxyl group of 1,2-diol). This compound was pre-
pared via oxidation of 1-(p-methoxyphenyl)-1,2-ethane-
diol. To a stirred suspension of p-methoxystyrene (4.0
g, 23.2 mmol) in water (130 mL), powdered m-CPBA
(2.0 g, 19.2 mmol) was added in small portions over a
period of 5–10 min at 0°C. The mixture was stirred at
rt for 2 h. Then 10% aqueous sulfuric acid (3.3 mL) was
added and the mixture was further stirred for 2 h. The
progress of the reaction was monitored by TLC. Solid
NaOH was added until the solution became homoge-
neous. The reaction mixture was extracted with ethyl
acetate (5×10 mL). The combined organic extract was
washed with brine, dried over anhydrous Na2SO4, and
concentrated in vacuo. The residue was purified by
silica-gel column chromatography. Elution with hex-
ane/ethyl acetate (1/1) afforded 1-p-methoxyphenyl-1,2-
According to the general procedure, 11c was prepared
from 1-(p-chlorophenyl)-1,2-ethanediol (500 mg, 2.88
mmol); yield 183 mg (37%); mp 116.0–118.0°C (from
ethanol), lit.12 114–115°C; IR wmax: 3424, 3382, 1680,
1
1
ethanediol 12 (474 mg, 15%); H NMR (270 MHz): l
3.72 (m, 2H), 3.83 (s, 3H), 4.79 (m, 1H), 6.91 (d,
1592, 1232, 1093, 827 cm−1; H NMR (270 MHz): l
4.83 (s, 2H), 7.46 (d, J=8.57 Hz, 2H), 7.85 (d, J=8.57
J=8.73, 2H), 7.34 (d, J=8.73, 2H).
Hz, 2H).
To a stirred solution of 1-p-methoxyphenyl-1,2-ethane-
diol 12 (500 mg, 2.97 mmol) and 4-dimethylaminopyr-
idine (14.5 mg, 0.12 mmol) in dichloromethane (15 mL)
was added triethylamine (331 mg, 3.27 mmol) at rt.
tert-Butyldimethylsilyl chloride (493 mg, 3.27 mol) was
added dropwise over 20 min and the stirring was con-
tinued overnight at rt. The progress of the reaction was
monitored by TLC. Removal of the solvent afforded
a residue, which was purified by silica-gel column
chromatography. Elution with hexane/ethyl acetate
(5/1) afforded p-methoxyphenyl)-2-tert-butyldimethyl-
silyloxyethanol (800 mg, 95%). Solid tetrapropyl-
ammonium perruthenate (TPAP, 46.7 mg, 0.13 mmol)
was added in one portion to a stirred solution of
1 - (p - methoxyphenyl) - 2 - tert - butyldimethylsilyloxy-
ethanol (750 mg, 2.66 mmol) and N-methylmorpholine-
N-oxide (MMO, 467 mg, 398 mmol) in dichloro-
methane at rt. The stirring was continued for 3 h at the
same temperature. Then the reaction mixture was
filtered through a pad of silica, eluting with ethyl
acetate. The filtrate was evaporated to afford the
residue, which was stirred in a 50% aqueous methanolic
solution of Oxone (1.8 g, 2.93 mmol) at rt for 3 h. The
progress of the reaction was monitored by TLC. On
completion, the methanol was removed under reduced
pressure and the residue was extracted five times with
10 mL portions of ethyl acetate. The combined organic
extract was washed with brine, dried over anhydrous
Na2SO4, and concentrated in vacuo. The residue was
purified by silica-gel column chromatography. Elution
with hexane/ethyl acetate (2/1) afforded p-methoxy-a-
hydroxyacetophenone 11b as colorless crystals; yield
386 mg (three steps, 83%); mp 105.0–105.5°C (from
ethanol), lit.11 105–106°C; IR wmax: 3418, 1681, 1673,
4.2.3. 1-Benzyloxy-3-hydroxy-2-propanone 15a. To a
stirred mixture of epichlorohydrin (2.0 g, 21.6 mmol),
benzyl alcohol (2.12 g, 19.6 mmol), and tetra-
butylammonium hydrogen sulfate (0.29 g, 0.86 mmol)
was added dropwise 50% aqueous sodium hydroxide
(10 mL) at 0°C. Stirring was continued for 30 min at
the same temperature and then for 21 h at rt. The
mixture was extracted with ether (5×10 mL). The com-
bined organic extract was washed with brine, dried over
anhydrous Na2SO4 and concentrated in vacuo. The
residue was purified by silica-gel column chromatogra-
phy. Elution with hexane/ether (6/1) afforded 1-benzyl-
oxy-2,3-epoxypropane; yield 1.84 g (57%); oil; 1H NMR
(270 MHz): l 2.54 (dd, J=4.94, 2.80, 1H), 2.72 (dd,
J=4.61, 4.45, 1H), 3.11 (m, 1H), 3.35 (dd, J=11.33,
5.77, 1H), 3.67 (dd, J=11.38, 2.96, 1H), 4.45 (d, J=
2.03, 1H), 4.53 (d, J=2.03, 1H), 7.22 (m, 5H).
A mixture of 1-benzyloxy-2,3-epoxypropane (1.0 g, 6.09
mmol) and 1% aqueous solution of sulfuric acid (30
mL) was stirred for 5 h at rt. The aqueous solution was
extracted with ethyl acetate (10×10 mL). The combined
organic extract was washed with brine, dried over anhyd-
rous Na2SO4 and concentrated in vacuo to give 1-benz-
1
yloxy-2,3-propanediol 16a; yield 1.06 g (96%); oil; H
NMR (270 MHz): l 3.59 (m, 4H), 3.83 (m, 1H), 4.49 (s,
2H), 7.27 (m, 5H).
According to the general procedure, 15a was prepared
from 1-benzyloxy-2,3-propanediol (16a) (500 mg, 2.74
mmol); yield 84 mg (17%); mp 77.0–78.0°C; IR wmax
:
3361, 3300, 2939, 2878, 1731, 1498, 1127, 1094 cm−1; 1H
NMR (270 MHz): l 4.20 (s, 2H), 4.50 (s, 2H), 4.62 (s,
2H), 7.37 (m, 5H).
1
1604, 1233, 836 cm−1; H NMR (270 MHz): l 3.87 (s,

T. Tsujigami et al. / Tetrahedron: Asymmetry 12 (2001) 2543–2549
2547
4.2.4. 1-Benzoyloxy-3-hydroxypropanone 15b. To
a
phosphate buffer (0.1 M, pH 6.5). The wet cells (4 g)
were re-suspended in 20 mL of a phosphate buffer
solution (pH 6.5, 0.1 M) in a 500 mL shaking culture
(Sakaguchi) flask with the substrate 11a [103 mg, 0.76
mmol, 0.5% w/v]. After glucose (1.0 g) was added, the
flask was purged with argon, equipped with a balloon
filled with argon and shaken on a reciprocal shaker
for 2 days at 30°C. The cell mass was removed by
filtration using Celite. The filtrate was extracted with
ethyl acetate (10×10 mL). The combined organic
extract was washed with brine and dried over anhyd-
rous Na2SO4. The solvent was removed in vacuo and
the residue was purified by silica-gel column chro-
matography. Elution with hexane/ethyl acetate (1/1)
afforded (S)-1-phenyl-1,2-ethanediol 12a (95 mg, 91%);
IR wmax: 3300–3200, 3030, 2933, 2871, 1448, 1101,
stirred mixture of dihydroxyacetone dimer (180 mg,
2.00 mmol) and benzoic anhydride (693 mg, 3.02
mmol) in 1,4-dioxane (20 mL), lipase PS (15 g) was
added, and the mixture was stirred for 48 h at 30°C.
After the removal of the enzyme by filtration with
Celite, the filtrate was concentrated under reduced
pressure. The residue was purified by silica-gel column
chromatography. Elution with hexane/ethyl acetate (1/
1) afforded 15b; yield 178.0 mg (46%); mp 94.0–95.0°C
(from ethanol), lit.13 92–93°C; IR wmax: 3430, 3382,
1730, 1714, 1273, 1110, 713 cm−1; 1H NMR (270
MHz): l 4.49 (s, 2H), 5.00 (s, 2H), 7.52 (m, 3H), 8.07
(m, 2H).
4.2.5. 1-Hydroxy-4-phenyl-2-butanone 17a. According
to the procedure for the preparation of 12, diol 18a
was prepared from 4-phenyl-1-butene (3 g, 23 mmol);
yield 3 g (80%); IR wmax: 3400–3100, 2930, 2862, 2361,
1
1054, 700 cm−1; H NMR (270 MHz): l 3.61 (m, 2H),
4.72 (m, 1H), 7.20 (m, 5H); MS m/e (%): 31 (110), 77
(80), 79 (75), 107 (20), 138 (M+, 2); [h]D22 +71.9 (c 1.04,
CHCl3), lit.14 [h]D18 +60 (c 1.0, CHCl3); mp 64.5–65.0°C
(from ethanol, lit.15 62–63°C. Its e.e. was determined
by HPLC analysis (Chiralcel OD of Daicel Chemical
Industries Ltd., 4.6×250 mm); eluent: hexane-2-
propanol=9:1; flow rate 0.5 mL/min, retention time:
(R)-form; 22.3 min (minor), (S)-form; 23.8 min
(major).
2339, 1496, 1455, 1097, 1069, 1038, 699, 519 cm−1; H
1
NMR (270 MHz): l 1.69–1.82 (m, 2H), 2.65–2.87 (m,
2H), 3.47 (m, 1H), 3.71 (m, 2H), 7.17–7.32 (m, 5H).
According to the general procedure, 17a was prepared
from ( )-4-phenyl-1,2-butanediol (18a, 967 mg, 5.82
mmol); yield 216 mg (23%); oil; IR wmax: 3500–3400,
3027, 2927, 2361, 2339, 1721, 1604, 1496, 1454, 1064,
1
1496, 1454, 1064, 993, 749, 701 cm−1; H NMR (270
4.3.2. (S)-(+)-1-(p-Methoxyphenyl)-1,2-ethanediol (S)-
12b. The cultivation of the Y. farinosa and the follow-
ing reaction were carried out according to the
procedure described for the preparation of (S)-12a.
1-Hydroxy-2-(4-methoxyphenyl)ethanone 11b (103 mg,
0.62 mmol) was used as the substrate; yield 99 mg
(95%); IR wmax: 3400–3300, 2936, 1612, 1515, 1245,
MHz): l 2.72 (d, J=7.75, 2H), 2.96 (d, J=7.75, 2H),
4.17 (s 2H), 7.14–7.27 (m, 5H).
4.2.6. 1-Hydroxy-3-phenoxypropanone 17b. According
to the procedure of the preparation of styrene glycol
derivatives 12, 1-phenoxy-2,3-propanediol 18b was pre-
pared from 1,2-epoxy-3-phenoxypropane (3 g, 20
mmol); yield 2.66 g (79%); mp 56.5–57.0 (from etha-
nol); IR wmax: 3450–3300, 2916, 2871, 2360, 2338,
1600, 1497, 1252, 1046, 748 cm−1; 1H NMR (270
MHz): l 4.02–4.11 (m, 3H), 6.88–6.99 (m, 3H), 7.23–
7.31 (m, 2H).
1
1086, 1025, 834 cm−1; H NMR (270 MHz) l 3.72 (m,
2H), 3.83 (s, 3H), 4.79 (m, 1H), 6.91 (d, J=8.73, 2H),
7.34 (d, J=8.73, 2H); MS m/e (%): 31 (46), 77 (71), 94
(73), 109 (85), 137 (100), 168 (M+, 18); [h]D23 +64.5 (c
1.02, CHCl3); mp 78.0–79.0°C (from ethanol), lit.16
75–76°C. Its e.e. was determined by HPLC analysis
(Chiralcel OD of Daicel Chemical Industries Ltd.,
4.6×250 mm; eluent: hexane-2-propanol=9:1; flow rate
0.5 mL/min, retention time: (R)-form; 29.8 min
(minor), (S)-form; 32.2 min (major).
According to the general procedure, 17b was prepared
from ( )-18b (1 g, 5.95 mmol); yield 309 mg (31%);
mp 147.5–148.0°C (from ethanol); IR wmax: 3375, 1749,
1738, 1706, 1601, 1587, 1499, 1249, 1076, 1053 cm−1;
1H NMR (270 MHz, acetone-d6): l 3.68 (d, J=11.54,
2H), 3.94 (d, J=11.71, 1H), 3.98 (d, J=11.71, 1H),
4.26 (d, J=11.54, 1H), 6.90–6.97 (m, 3H), 7.25–7.31
(m, 2H).
4.3.3. (S)-(+)-1-(4-Chlorophenyl)-1,2-ethanediol (S)-12c.
The cultivation of the Y. farinosa and the following
reaction were carried out according to the procedure
described for the preparation of (S)-12a. 2-(4-
Chlorophenyl)-1-hydroxyethanone (11c, 61 mg, 0.36
mmol) was used as the substrate; yield 52 mg (85%);
IR wmax: 3400–3300, 2933, 2895, 1492, 1087, 1037, 900,
4.3. General procedure for the microbial reductions
1
The substrates were added to the suspension of grown
cells of Yamadazyma farinosa IFO 10896 and incu-
bated at 30°C.
829 cm−1; H NMR (270 MHz): l 3.60 (dd, J=11.21,
8.08 Hz, 1H), 3.74 (dd, J=11.21, 3.46 Hz, 1H), 4.79,
(dd, J=8.08, 3.46 Hz, 1H), 7.30 (m, 4H); MS m/e (%):
31 (40), 77 (100), 141 (45), 143 (15), 172 (M+, 2.6), 174
(0.9); [h]2D2 +59.5 (c 1.07, CHCl3) [lit.17 [h]2D0 −31.4 for
the (R) enantiomer (c 0.97, EtOH)]; mp 80.0–80.5°C
(lit.17 83–84°C). Its e.e. was determined by HPLC
analysis (Chiralcel OD of Daicel Chemical Industries
Ltd., 4.6×250 mm; eluent: hexane-2-propanol=9:1;
flow rate 0.5 mL/min, retention time: (R)-form; 36.6
min (minor), (S)-form; 39.6 min (major).
4.3.1. (S)-(+)-1-Phenyl-1,2-ethanediol (S)-12a. Y. fari-
nosa IFO 10896 was incubated in a glucose medium
[containing glucose (2.0 g), peptone (0.7 g), yeast
extract (0.5 g), K2HPO4 (0.2 g), KH2PO4 (0.3 g), pH
6.5, total volume 100 mL] for 2 days at 30°C. The wet
cells (ca. 8 g from 100 mL of the broth) were har-
vested by centrifugation (3000 rpm) and washed with

2548
T. Tsujigami et al. / Tetrahedron: Asymmetry 12 (2001) 2543–2549
4.3.4. (R)-1-Benzyloxy-2,3-propanediol (R)-16a. The
cultivation of the Y. farinosa and the following reaction
were carried out according to the procedure described
for the preparation of (S)-12a. 1-Benzyloxy-3-hydroxy-
propan-2-one 15a (99 mg, 0.55 mmol) was used as the
substrate; yield 98 mg (98%); IR wmax: 3450–3350, 2929,
3H), 7.23–7.31 (m, 2H); MS m/e (%): 31 (100), 39 (64),
51 (54), 65 (66), 77 (26), 95 (21), 168 (M+, 40); [h]D20
−1.61 (c 1.02, CHCl3), lit.21 [h]2D0 −10.8 (c 1.0 EtOH) for
the (R) isomer; mp 56.5–57.0°C (from ethanol), lit.22
53.5–55°C. Its e.e. was determined by HPLC analysis
(Chiralcel OD of Daicel Chemical Industries Ltd., 4.6×
250 mm; eluent: hexane-2-propanol=9:1; flow rate 2.0
mL/min, retention time: (R)-form; 9.7 min (major),
(S)-form; 21.6 min (minor).
1
2869, 1736, 1107, 1045, 699 cm−1; H NMR (270 MHz)
l 3.59 (m, 4H), 3.83 (m, 1H), 4.49 (s, 2H), 7.27 (m, 5H);
MS m/e (%): 31 (25), 77 (232), 91 (100), 105 (49), 182
(M+, 4.4); [h]D21 −0.98 (c 1.02, CHCl3) lit.18 [h]D20 −1.9 (c
0.84, CHCl3) for the (R) enantiomer. Its e.e. was deter-
mined by HPLC analysis (Chiralcel OD of Daicel
Chemical Industries Ltd., 4.6×250 mm; eluent: hexane-
2-propanol=9:1; flow rate 1.0 mL/min, retention time:
(R)-form; 18.5 min (major), (S)-form; 22.9 min
(minor).
Acknowledgements
This work was partially supported by the Grant-in-Aid
for Scientific Research (10490025) from the Ministry of
Education, Culture, Sports, Science, and Technology,
Japan. This work was also supported by the ‘Research
for the Future Program (JSPS-RFTF 90100302)’ from
the Japan Society for the Promotion of Science.
4.3.5. (S)-1-Benzoyloxy-2,3-propanediol (S)-16b. The
cultivation of the Y. farinosa and the following reaction
were carried out according to the procedure described
for the preparation of (S)-12a. 1-Benzoyloxy-3-hydroxy-
propanone (15b, 103 mg, 0.53 mmol) was used as the
substrate; yield 68 mg (65%); IR wmax: 3450–3350, 2951,
2888, 1719, 1602, 1452, 1316, 1278, 1107, 1070, 712
References
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cm−1; H NMR (270 MHz): l 3.71 (dd, J=11.46, 5.69,
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17a (99 mg, 0.60 mmol) was used as the substrate; yield
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2339, 1721, 1604, 1496, 1454, 1064, 1496, 1454, 1064,
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(S)-form; 11.0 min (major), (R)-form; 24.6 min
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