Candida tropicalis CE017: A new Brazilian enzymatic source for the bioreduction of aromatic prochiral ketones

Article abstract of DOI:10.1590/S0103-50532010000800015

The reactivity and stereoselectivity showed by a new strain of Candida tropicalis in the reduction of prochiral ketones have been compared with the ones previously attained in our laboratory using microorganisms from the Brazilian biodiversity. In this manner, Candida tropicalis has demonstrated its versatility as stereoselective agent in the bioreduction of a series of aromatic ketones. These prochiral compounds were converted into their corresponding optically alcohols with moderate to excellent stereopreference depending on the substrate structure. Among ketones tested, nitroacetophenones were enzymatically reduced to enantiopure (S)-alcohol with complete conversion.

Full text of DOI:10.1590/S0103-50532010000800015

J. Braz. Chem. Soc., Vol. 21, No. 8, 1509-1516, 2010.
Printed in Brazil - ©2010 Sociedade Brasileira de Química
0103 - 5053 $6.00+0.00
Candida tropicalis CE017: a New Brazilian Enzymatic Source for the Bioreduction
of Aromatic Prochiral Ketones
Gizelle A. B. Vieira,a Daniel M. de Freitas Araujo,a Telma L. G. Lemos,a
Marcos Carlos de Mattos,*,a Maria da Conceição F. de Oliveira,a Vânia M. M. Melo,a
Gonzalo de Gonzalo,b Vicente Gotor-Fernándezb and Vicente Gotor*,b
aDepartamento de Química Orgânica e Inorgânica, Universidade Federal do Ceará,
60451-970 Fortaleza-CE, Brazil
bDepartamento de Química Orgánica e Inorgánica, Instituto Universitario de Biotecnología de
Asturias, Universidad de Oviedo, c/ Julián Clavería 8, 33006, Oviedo, Spain
A reatividade e estereosseletividade apresentadas por uma nova cepa de Candida tropicalis
na redução de cetonas pró-quirais foram comparadas com cepas de microrganismos, estudadas
em nosso laboratório, provenientes da biodiversidade brasileira. Neste caso, Candida tropicalis
demonstrou-se como um agente redutor versátil e estereosseletivo na biorredução de uma série de
cetonas aromáticas. Estes compostos pró-quirais foram convertidos aos correspondentes álcoois
opticamente ativos com moderada a excelente estereopreferência dependendo da estrutura do
substrato. Entre as cetonas utilizadas, as nitroacetofenonas foram enzimaticamente reduzidas aos
correspondentes (S)-álcoois enantiopuros com completa conversão.
The reactivity and stereoselectivity showed by a new strain of Candida tropicalis in the
reduction of prochiral ketones have been compared with the ones previously attained in our
laboratory using microorganisms from the Brazilian biodiversity. In this manner, Candida tropicalis
has demonstrated its versatility as stereoselective agent in the bioreduction of a series of aromatic
ketones. These prochiral compounds were converted into their corresponding optically alcohols
with moderate to excellent stereopreference depending on the substrate structure. Among ketones
tested, nitroacetophenones were enzymatically reduced to enantiopure (S)-alcohol with complete
conversion.
Keywords: alcohol dehydrogenases, biocatalysis, Candida tropicalis, ketones, yeast
Introduction
methods usually involves the use of expensive reagents or
heavy metal catalysts.3 By contrast, biocatalysis applied to
industrial processes has been shown as a very advantageous
alternative to conventional chemical methods, and is
widely used for the preparation of enantiomerically pure
pharmaceuticals and other high added value compounds.4
Enzymatic reduction of carbonyl groups represents
one of the most important reactions employed in the
synthesis of chiral alcohols. Enzymes that can be used
for this transformation are oxidoreductases, which
require the presence of a coenzyme, such as NADH or
NADPH, which transfers the hydride anion to the carbonyl
compound, being formed NAD+ or NADP+. Whole cells of
microorganisms can be also used in the enzymatic reduction
of the carbonyl group. Each cell represents a small factory
fully equipped for the reduction of the substrate bearing
Chiral alcohols are an important class of organic
substrates due to their properties as bioactive compounds
and as starting materials for the synthesis of various
biologically active materials.1 For example, enantiopure
1-phenylethanol is a chiral building block used in the
fine chemical and pharmaceutical industries as an
ophthalmic preservative, a solvatochromic dye, an inhibitor
of cholesterol intestinal absorption and a mild floral
fragrance.2 In the synthesis of enantiopure chiral secondary
alcohols, prochiral ketones are commonly used as starting
materials using stereoselective chemical transformations or
biocatalytic methods. Asymmetric reduction by chemical
*e-mail: mcdmatto@ufc.br; vgs@fq.uniovi.es

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Candida tropicalis CE017: a New Brazilian Enzymatic Source
J. Braz. Chem. Soc.
the carbonyl moiety. In order to find optimal conditions
for the action of cell factories, the process designer has
only to plan in which way the ketonic substrate should be
added, and which medium should be used for the designed
reaction. Nowadays, a wide variety of cultured cells from
microorganisms or vegetables are available for enzyme-
mediated reduction of carbonyl compounds.5
purities (97-99% ee) in all cases, observing the maximum
conversion (62%, entry 4) after 9 days of reaction. Longer
reaction times led to the formation of acetophenone due to
an oxidizing activity showed by Candida tropicalis.
Table 1. Effect of incubation time on the reduction of acetophenone by
C.tropicalis CE017 at 28 ºC and pH 7.0
It is well known that the screening of a wide variety of
microorganisms living in the environment is an efficient
method to obtain the desired enzyme towards an unnatural
substrate.Acetophenone has been used as a model substrate
for the isolation of microorganisms having ketone reductase
activity.6 Recently, a new strain of Candida tropicalis
PBR-2 MTCC 5158 has been reported as an efficient
reducing agent to acetophenone and its derivatives.7
The use of microorganism new strains as biocatalysts
may offer an alternative opportunity to investigate the
local resources for the effective conduct of key synthetic
transformations with significant economic and ecological
implications.8
Our research interest is based on the development of
chemoenzymatic methodologies to obtain chiral alcohols
using Brazilian local sources of low cost. Herein, we report
our current investigations in the study of the Candida
tropicalis CE017 strain behavior as a novel stereoselective
reducing agent of aromatic prochiral ketones to the
corresponding chiral alcohols. The experimental results
obtained will be compared to the one previously obtained
by our research group using different Brazilian enzymatic
sources.9,10
Entry
time / days
Conversion / (%)a
ee / (%)b
> 99 (S)
98 (S)
1
2
3
4
5
6
1
3
20
35
38
62
60
56
6
97 (S)
9
97 (S)
12
15
97 (S)
97 (S)
aConversion was determined by GC. bDetermined by GC. By comparison
with (S)- and (R)-1-phenylethanol standards was possible to verify that
the major enantiomer formed in the enzymatic process possesses the S
configuration.
In our efforts to improve the yield of (S)-1-phenylethanol
1b, some other reaction parameters were analyzed as culture
medium, pH, temperature and substrate concentration.
Culture medium
Acetophenone bioreductions were performed during
9 days, at 28 ºC, using two different culture media to
obtain whole cells of C. tropicalis CE017: (i) peptone
10 g L-1, dextrose 20 g L-1 pH 6.5,11 and (ii) Czapeck:
NaNO3 (3 g L-1), K2HPO4 (1.0 g L-1), MgSO4.7H2O, KCl
(0.5 g L-1), FeSO4 (0.01 g L-1), dextrose (30 g L-1), pH 8.5.12
Additionally the bioreduction was performed adjusting the
pH of both media to 5.5 with a sterile solution of tartaric
acid 10%. The results obtained are listed in Table 2.
Results and Discussion
Bioreduction of acetophenone using C. tropicalis CE017
Table 2. Effect of culture medium on the bioreduction of acetophenone
by C. tropicalis CE017
Acetophenone 1a was tested in the bioreduction using
growing cells of the yeast as biocatalyst in a potato dextrose
broth medium.
Entry Culture medium
pH
5.5
6.5
8.5
5.5
Conversion / (%)a
ee / (%)a
99 (S)
99 (S)
99 (S)
99 (S)
1
2
3
4
Peptone/Dextrose
Peptone/Dextrose
Czapeck
13
13
11
12
O
OH
Growing cells of
Candida tropicalis
PD medium
125 rpm, 28 ºC
Czapeck
aConversion and enantiomeric excesses were determined by GC.
1a
(S)-1b
Scheme 1. Bioreduction of acetophenone 1a by C. tropicalis.
These results revealed that all the new culture media
have similar effect in the enzymatic activity (11-13%
conversion), allowing the recovery of (S)-1-phenylethanol
in enantiomerically pure form. All these media led to
notable lower isolated yields in comparison with potato
dextrose medium (62% yield at pH 7.0).
Enzymatic reductions have been followed by analyzing
the product 1b formed after 1, 3, 6, 9, 12 and 15 days
of reaction, being summarized the data in Table 1.
(S)-1-phenylethanol 1b was obtained with high optical

Vol. 21, No. 8, 2010
Vieira et al.
1511
pH Influence
(S)-1-phenylethanol in high to excellent enantiomeric
excess (Table 5).
Reactions were analyzed after 9 days using potato
dextrose broth as culture medium in the following pHs:
4.0, 5.5, 7.0 and 8.0. The medium was adjusted to pH 8.0
and 7.0 with a sterile solution of NaOH 1.0 mol L-1, and
pH 4.0 was adjusted by using a sterilized 10% tartaric acid
solution. The results were summarized in Table 3.
A slight improvement was observed in the conversion
when less quantity of substrate (6-12 μL) was used, but
in these conditions was observed a 85% ee in (S)-1-
phenylethanol (entries 1-4). The conversion decreased
when more quantity of substrate was used (entries 5-9),
however higher concentrations of ketone led to better
stereopreference values, being in this case 15-20 μL the
appropriate quantity of ketone (entries 6-7).
Table 3. Effect of pH on the bioreduction of acetophenone by C. tropicalis
CE017
Thus, a temperature of 28 ºC, 9 days of reaction time,
pH of 5.5, potato-dextrose broth as culture medium and
20 μL of acetophenone proved to be the optimum conditions
to obtain (S)-1-phenylethanol (1b) in good yield (62%) and
high enantiomeric excess (97% ee).
Entry
pH
4.0
5.5
7.0
8.0
Conversion / (%)a
ee / (%)a
98 (S)
98 (S)
98 (S)
98 (S)
1
2
3
4
50
60
62
42
Table 5. Effect of the quantity of substrate on the bioreduction of
acetophenone (1a) by C. tropicalis CE017
aConversion and enantiomeric excesses (ee) were determined by GC.
Entry Quantity of 1a / μL
Conversion / (%)a
ee / (%)a
85
The optimal pH for this biocatalytic system varied
from 5.5 to 7.0 (entries 2 and 3), observing a decrease of
conversion when more acidic or basic media were employed
(entries 1 and 4).
1
2
3
4
5
6
7
8
9
6
75
75
71
72
61
60
62
48
44
8
85
10
12
14
15
20
30
40
85
85
Temperature effect on Candida tropicalis
92
97
Reactions were performed using potato dextrose broth
at higher temperatures (45 and 55 ºC), as shown in Table 4.
97
91
94
Table 4. Effect of temperature on the bioreduction of acetophenone by
C. tropicalis CE017
a Conversion and enantiomeric excesses (ee) were determined by GC.
Entry
Temperature /
Days
Conversion /
(%)a
ee /
(%)a
Bioreduction of acetophenone derivatives
(oC)
1
2
3
45
45
55
3
9
9
16
51
14
97 (S)
97 (S)
68 (S)
After these encouraging results, we decided to extend
our studies toward the analysis of the bioreduction
of substituted acetophenones. Thus, ketones 2a-10a
(Scheme 2) were reduced by growing cells of C. tropicalis
CE017 using the optimal reaction conditions attained in the
bioreduction of acetophenone. The results are summarized
in Table 6.
The influence of an electron-donor (OMe) group and
an electron-withdrawing (NO2) group in ortho, meta- and
para- position of the aromatic moiety was analyzed.
Reduction of nitroacetophenones occurred with complete
conversion yielding the corresponding alcohols (S)-
2b-4b with excellent enantioselectivities (entries 1-3).
The presence of the nitro group strongly increased the
enzymatic activity when compared with acetophenone.
Ketone 5a having the methoxy group in the ortho position
yielded the corresponding alcohol (S)-5b with moderate
aConversion and enantiomeric excesses (ee) were determined by GC.
These results revealed that C. tropicalis does not
present a good thermostabilitity, as a decrease of the ketone
reductase activity was observed by working at temperatures
above 28 ºC (entries 1 and 2), leading to a dramatic loss of
yield and also of the enantiomeric excess especially over
45 ºC (entry 3).
Quantity of substrate
Bioreduction reactions were performed during 9
days at 28 ºC in potato dextrose broth using 6, 8, 10, 12,
14, 15, 20, 30 and 40 μL of acetophenone 1a to obtain

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Candida tropicalis CE017: a New Brazilian Enzymatic Source
J. Braz. Chem. Soc.
O
OH
*
R1
R1
Candida tropicalis CE017
Potato Dextrose medium
125 rpm / 28 ºC
R4
R2
R4
R2
9 days
R3
2a-10a
R3
(S)- or (R)-2b-10b
7 R1 = H R2 = H R3 = H R4 = OMe
8 R1 = H R2 = H R3 = H R4 = Cl
9 R1 = Cl R2 = H R3 = H R4 = H
10 R1 = Cl R2 = H R3 = Cl R4 = Cl
2 R1 = H R2 = NO2 R3 = H
R4 = H
3 R1 = H R2 = H
4 R1 = H R2 = H
R3 = NO2 R4 = H
R3 = H
R4 = NO2
R4 = H
R3 = OMe R4 = H
5 R1 = H R2 = OMe R3 = H
6 R1 = H R2 = H
Scheme 2. Bioreduction of prochiral aromatic ketones 2a-10a using C. tropicalis CE017.
Table 6. Candida tropicalis CE017 biocatalyzed reduction of ketones 2a-10a in potato-dextrose medium at 28 °C, 125 rpm and 9 days
Entry
Ketone
2a
R1
H
H
H
H
H
H
H
Cl
Cl
R2
NO2
H
R3
H
R4
H
Conversion / (%)a
ee / (%)a
> 99 (S)
> 99 (S)
96 (S)
56 (S)
80 (S)
9 (S)
1
2
3
4
5
6
7
8
9
> 99
> 99
> 99
51
3a
NO2
H
H
4a
H
NO2
H
5a
OMe
H
H
6a
OMe
H
H
35
7a
H
OMe
Cl
7
8a
H
H
65
62 (S)
89 (R)
90 (R)
9a
H
H
H
12
10a
H
Cl
Cl
63
aConversion and enantiomeric excesses (ee) were determined by HPLC or GC (see Experimental section). Absolute configuration of the optically alcohol
in brackets.
conversion and selectivity (entry 4). When the methoxy
group was located at meta position, the conversion was
lower when compared with ketone 5a, being achieved
a higher selectivity (entry 5). A strongly decrease in the
activity and selectivity was observed for the reduction of
4-methoxyacetophenone (entry 6). The presence of the
methoxy group in the para position seemed to be worst
for enzymatic activity and selectivity.
4-Chloroacetophenone 8a yielded the (S)-8b with
moderate conversion and selectivity (entry 7). The presence
of the chlorine atom in the α-position to the carbonyl group
produced a strong decrease in the product conversion
with a good enantioselectivity, as shown in entry 8 for
the preparation of alcohol (S)-9b, yielding in this case the
(R)-alcohol due to CIP rules (entry 8).
Bioreduction of 10a, compound presenting two
chlorine atoms in the aromatic ring and one chlorine atom
in the α-position to the carbonyl group, occurred with
a similar conversion to acetophenone 8a and a similar
stereoselectivity compared to ketone 9a (entry 9). Again,
(R)-10b was obtained due to a change of priority in the
CIP rules.
Our research group has been working to select several
microorganisms from Brazilian biodiversity in order to
perform selective bioreduction reactions using whole
cells. We have found some interesting suppliers of alcohol
dehydrogenases such Lasiodiplodia theobromae9 and
Lentinus strigellus10 and now we can compare the results
previously obtained in these manuscripts with Candida
tropicalis. Each one is worth of attention due to their
particularities. Candida tropicalis has not a good affinity for
acetophenones bearing methoxy group moiety. Opposite,
fungal cells of L. theobromae and L. strigellus catalyze
the bioreduction of these methoxyacetophenones with
higher enantioselectivities (87-99%). On the other hand,
L. theobromae had poor selectivity for nitroacetophenones,
3a (only 45% of ee) and 4a (42% of ee). Lentinus strigellus
had good enantioselectivities (98-99%) but moderate
conversion of 2a (48%), that could not be improved after
longer reaction times. Candida tropicalis was the most
successful among them to reduce nitroacetophenones with
high to excellent conversion and enantioselectivity values.
It must be emphasize that nitrocompounds are important
building blocks in organic synthesis13 as well as precursors

Vol. 21, No. 8, 2010
Vieira et al.
1513
of the corresponding amines after a reduction process.14 In
addition, alcohol 6b is a relevant compound in the synthesis
of (S)-Rivastigmine15 and the alcohol 10b can be used in
the preparation of the pharmacological active Sertraline.16
In conclusion, all this family of chiral alcohols can be
obtained by an enzymatic strategy, just selecting the best
local microorganism in each specific case.
Varian for determining the optical purities. 1H, 13C NMR,
DEPT, and 1H-13C heteronuclear experiments were obtained
using AC-200 (1H, 200.13 MHz and 13C, 50.3 MHz),
AC-300 (1H, 300.13 MHz and 13C, 75.5 MHz), DPX-300
(1H, 300.13 MHz and 13C, 75.5 MHz) or AV-400 (1H,
400.13 MHz and 13C, 100.6 MHz) Bruker spectrometers.
The chemical shifts are given in delta (d) values and the
coupling constants (J) in Hertz (Hz). Optical rotations were
performed in a Perkin-Elmer 241 polarimeter.
Conclusions
Growing cells of a new yeast strain, C. tropicalis CE017
in potato dextrose have been shown as an excellent and
inexpensive biocatalyst for the introduction of chirality in
bioreduction processes.When reducing substituted aromatic
ketones, the presence of a nitro group in the aromatic
moiety has a positive effect in the biocatalytic properties of
C. tropicalis CE017, yielding the corresponding alcohols
with complete conversion and ee > 95%. On the other hand,
methoxyacetophenones seemed to be poorer substrates for
this yeast. In all cases the biocatalyst has presented Prelog
selectivity for the production of optically active alcohols.
Comparison with two microorganisms previously studied
in our group (L. theobromae and L. strigellus) revealed its
particularities. In this case, Candida tropicalis was the most
successful among them to reduce nitroacetophenones with
great conversion and enantioselectivity. The evaluation of
locally available microorganisms for a selection of standard
organic chemical reactions of commercial significance
could prove to be a very valuable economic endeavor.
It may well offer new opportunities to expand the local
resources as sustainable enzymatic systems where high-
cost, unstable coenzymes are presently used.
General procedure for preparation of racemic alcohols
1a-10a
To a solution of the corresponding ketone 1a-10a
(200 mg) in dry MeOH (4.0 mL) was slowly added sodium
borohydride (4 equiv) at 0 ºC under nitrogen atmosphere.
The reaction was stirred at room temperature during 3 h
and followed by TLC analysis (20% EtOAc/hexane) until
complete disappearance of the starting ketone. Solvent
was evaporated under reduced pressure and the resulting
suspension was redissolved in H2O and extracted with
EtOAc (3 × 100 mL). Organic phases were combined and
dried over anhydrous Na2SO4. After solvent distillation
under reduced pressure, the resulting crude product was
purified by flash chromatography [10% EtOAc/hexane for
1b (88% yield) and 10b (92% yield), 15% EtOAc/hexane
for 5b (74% yield), 6b (86% yield), 7b (96% yield), 8b
(83% yield), 9b (90% yield) and 20% EtOAc/hexane for
2b (97% yield), 3b (95% yield), and 4b (95% yield)].
Candida tropicalis strain CE017
The strain CE017 was previously isolated from
an oil refinery wastewater located in Fortaleza city
(Ceara state, Brazil) by enrichment cultivation.17 This
strain was identified as Candida tropicalis according to
micromorphological and biochemical characteristics.18 It
is noteworthy that C. tropicalis CE017 showed a particular
ability to degrade high phenol concentration and might be
useful in bioremediation activities.17 This strain is deposited
in the Culture Collection of the Microbial Ecology and
Biotechnology Laboratory at Biology Department of the
Federal University of Ceará, Brazil.
Experimental
General
All reagents were purchased from Aldrich and used
without further purification. Dry solvents were distilled
over an adequate desiccant under nitrogen. Flash
chromatographies were performed using silica gel 60
(230-240 mesh). High performance liquid chromatography
(HPLC) analysis were carried out in a Hewlett Packard
1100 chromatograph UV detector at 210 nm using a Daicel
Chiralcel OB-H, Chiralpak AS or Chiralpak IA column
(25 cm × 4.6 mm I.D.) varying the conditions depending
on the specific substrate, and using mixtures of hexane
and 2-propanol (IPA). Gas chromatography (GC) analysis
were performed on a Hewlett Packard 6890 Series II
chromatograph equipped with a CP-Chiralsil DEX CB or a
HP-1 column (30 m × 0.25 mm × 0.25 μm, 1.0 bar N2) from
Culture conditions
For inoculums preparation, stock cultures were
cultivated in potato broth (Potato 4 g L-1, Dextrose 20 g L-1,
HIMEDIA) pH 5.5 for 24 h at 28 ºC under shaking
conditions (125 rpm). After this, the absorbance of the
culture was adjusted with 0.15 mol L-1 NaCl solution to

1514
Candida tropicalis CE017: a New Brazilian Enzymatic Source
J. Braz. Chem. Soc.
0.5 at 600 nm to obtain a suspension of 32 × 106 CFU mL-1
determined by counting of colony forming units (CFU) on
PotatoAgar Plates.An aliquot of 1.0 mL of this suspension
was inoculated in 250 mL Erlenmeyer flask containing
100 mL of Potato broth to produce biomass. When Peptone/
Dextrose was used as medium the counting of CFU was
15 × 106 CFU mL-1. In the case of Czapeck medium the
CFU value was 57 × 103 CFU mL-1.
tR 23.9 (S) min. Column AS. [α]D25 = +29.8, c 0.4, CHCl3,
for 99% ee of (S)-enantiomer.21
(S)-1-(3-Nitrophenyl)ethanol, 3b
1H NMR (CDCl3, 300.13 MHz): d 1.47 (3H, d, J 6.6 Hz,
H2), 4.96 (1H, q, J 6.6 Hz, H1), 7.47 (1H, t, J 7.8 Hz, H5’),
7.67 (1H, d, J 7.8 Hz, H6’), 8.04 (1H, ddd, J 8.3, 2.1 and
0.9 Hz, H4’), 8.18 (1H, t, J 1.8 Hz, H2’); 13C NMR (CDCl3,
75.5 MHz): d 25.2 (C2), 69.1 (C1), 120.2 (C4’), 122.1 (C2’),
129.3 (C5’), 131.6 (C6’), 147.8 (C1’), 148.1 (C3’). Conditions
for determination of the conversion by GC: Injector 225 oC,
Detector 250 oC, 80 oC (5 oC min-1) 180 oC (0 min), tR (3a)
9.5 min and tR (3b) 11.4 min. Column HP-1. Conditions
for determination of the ee by HPLC: 0.8 mL min-1,
hexane:IPA (90:10), 20 ºC, tR 12.8 min (S) and tR 14.2 (R)
min. Column OB-H. [α]D25 = -34.9, c 0.5 CH2Cl2, for 99%
ee of (S)-enantiomer.22
Procedure for the bioreduction of acetophenone and its
derivatives using Candida tropicalis
The growing cells of C. tropicalis CE017 were
used for bioreduction reactions according literature
procedure.19 Then, 20 mg of substrate were added into
erlenmeyer flask and the reactions were shaken for 9
days of reaction. The content of each flask was saturated
with sodium chloride, and then the aqueous phase was
extracted with EtOAc (3 × 80 mL). The organic phase
was dried over with Na2SO4 and then the solvent was
evaporated under reduced pressure. The reaction crude
was analyzed by the appropriate condition and purified by
flash chromatography. All the bioreduction experiments
were done in triplicate.
(S)-1-(4-Nitrophenyl)ethanol, 4b
1H NMR (CDCl3, 300.13 MHz): d 1.50 (3H, d, J 6.6 Hz,
H2), 5.01 (1H, q, J 6.6 Hz, H1), 7.53 (2H, d, J 8.7 Hz,
H2’+ H6’), 8.17 (2H, d, J 8.7 Hz, H3’ + H5’); 13C NMR
(CDCl3, 75.5 MHz): d 25.3 (C2), 69.5 (C1), 123.6
(C2’ + C6’), 126.0 (C3’ + C5’), 147.0 (C1’), 153.1 (C4’).
Conditions for determination of the conversion by GC:
Injector 225 oC, Detector 250 oC, 80 oC (5 oC min-1) 180 oC
(0 min), tR (4a) 9.6 min and tR (4b) 11.8 min. Column
HP-1. Conditions for determination of the ee by HPLC:
0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR 11.7 min (R) and
tR 12.8 (S) min. Column AS. [α]D25 = -24.6, c 0.5, CHCl3,
for 96% ee of (S)-enantiomer.23
(S)-1-Phenylethanol, 1b
1H NMR (CDCl3, 300.13 MHz): d 1.39 (d, J 6.5 Hz, 3H,
H2), 2.03 (s, 1H, OH), 4.77 (q, J 6.5 Hz, 1H, H1), 7.25 (m,
5H, H2’ + H3’ + H4’ + H5’); 13C NMR (CDCl3, 75.5 MHz):
d 25.3 (C2 ); 70.5 (C1), 125.5 (C2’ + C6’), 127.6 (C4’), 128.6
(C3’ + C5’), 146.0 (C1’). Conditions for determination of
the conversion by HPLC: 0.8 mL min-1, Hexane:IPA
(95:5), 20 ºC, tR (1a) 5.8 min and tR (1b) 8.4 min. Column
Spherisorb. Conditions for determination of the ee by
HPLC: 0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR 10.2 (S)
and tR 15.4 (R) min. Column OB-H. [α]D25 = -47.0, c 1.1,
CHCl3, for 97% ee of (S)-enantiomer. lit.: [α]D22 = -62.8
(c 1.0, CHCl3), 98.5% ee.20
(S)-1-(2-Methoxyphenyl)ethanol, 5b
1H NMR (CDCl3, 300.13 MHz): d 1.52 (3H, d, J 6.6 Hz,
H2), 3.88 (3H, s, OCH3), 5.11 (1H, q, J 6.6 Hz, H1), 6.90
(1H, dd, J 7.5 and 1.5 Hz, H3’), 6.98 (1H, dt, J 8.5 and
1.5 Hz, H5’), 7.29 (1H, dt, J 8.5 and 1.5 Hz, H6’), 7.36 (1H,
dd, J 7.5 and 1.5 Hz, H4’); 13C NMR (CDCl3, 75.5 MHz):
d 22.8 (C2), 55.2 (OCH3), 66.3 (C1), 110.3 (C3’), 120.7
(C5’), 126.0 (C6’), 128.2 (C4’), 133.4 (C1’), 156.4 (C2’).
Conditions for determination of the conversion by HPLC:
0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR (5a) 6.3 min
and tR (5b) 7.1 min. Column Spherisorb. Conditions for
determination of the ee by HPLC: 0.8 mL min-1, hexane:IPA
(95:5), 20 ºC, tR 10.3 (S) and tR 16.8 (R) min. Column OB-H.
[α]D25 = -4.5, c 0.4, CH2Cl2, for 56% ee of (S)-enantiomer.24
(S)-1-(2-Nitrophenyl)ethanol, 2b
1H NMR (CDCl3, 300.13 MHz): d 1.55 (3H, d, J 6.3
Hz, H2), 5.40 (1H, q, J 6.3 Hz, H1), 7.41 (1H, dt, J 8.1 and
1.2 Hz, H4’), 7.63 (1H, dt, J 8.1 and 1.5 Hz, H5’), 7.82 (1H,
dd, J 8.1 and 1.2 Hz, H6’), 7.88 (1H, dd, J 8.1 and 1.5 Hz,
H3’); 13C NMR (CDCl3, 75.5 MHz): d 24.2 (C2), 65.4 (C1),
124.2 (C3’), 127.5 (C6’), 128.0 (C4’), 133.5 (C5’), 140.9
(C1’ + C2’). Conditions for determination of the conversion
by GC: Injector 225 oC, Detector 250 oC, 80 oC (5 oC min-1)
180 oC (0 min), tR (2a) 8.4 min and tR (2b) 9.2 min. Column
HP-1. Conditions for determination of the ee by HPLC:
0.8 mL min-1, hexane:IPA (97:3), 20 ºC, tR 22.1 min (R) and
(S)-1-(3-Methoxyphenyl)ethanol, 6b
1H NMR (CDCl3, 300.13 MHz): d 1.47 (3H, d, J 6.6 Hz,
H2), 3.81 (3H, s, OCH3), 4.83 (1H, q, J 6.6 Hz, H1), 6.81
(1H, dd, J 8.5 and 1.2 Hz, H6’), 6.93 (2H, m, H2’ + H4’),

Vol. 21, No. 8, 2010
Vieira et al.
1515
7.26 (1H, t, J 8.5 Hz, H5’); 13C NMR (CDCl3, 75.5 MHz):
d 25.0 (C2), 55.1 (OCH3), 70.0 (C1), 110.8 (C2’), 112.7 (C4’),
117.6 (C6’), 129.3 (C5’), 147.6 (C1’), 159.6 (C3’). Conditions
for determination of the conversion by HPLC: 0.8 mL min-1,
hexane:IPA (95:5), 20 ºC, tR (6a) 5.1 min and tR (6b)
5.9 min. Column Spherisorb. Conditions for determination
of the ee by HPLC: 0.8 mL min-1, hexane:IPA (95:5),
20 ºC, tR 20.1 min (S) and tR 29.5 min (R). Column OB-H.
[α]D25 =-21.6, c0.34, MeOH, for80% eeof(S)-enantiomer.25
(R)-2-Chloro-1-(3,4-dichlorophenyl)ethan-1-ol, 10b27
1H NMR (CDCl3, 300.13 MHz): d 3.59 (1H, dd, J 11.4,
and 8.7 Hz, H2), 3.72 (1H, dd, J 11.4 and 3.6 Hz, H2),
4.87 (1H, dd, J 8.7 and 3.6 Hz, H1), 7.21 (1H, dd, J 8.4
and 1.8 Hz, H6’), 7.43 (1H, d, J 8.4 Hz, H5’), 7.51 (1H, d,
J 1.8 Hz, H2’); 13C NMR (CDCl3, 75.5 MHz): d 50.4 (C2),
72.8 (C1), 125.4 (C6’), 128.1 (C2’), 130.6 (C5’), 132.4 (C4’),
132.8 (C3’), 140.0 (C1’). Conditions for determination of
the conversion by GC: Injector 225 ºC, Detector 250 ºC,
80 ºC (5 ºC min-1) 180 ºC (0 min); tR (10a) 13.6 and tR (10b)
14.8 min. Column HP-1. Conditions for determination
of the ee by HPLC: 0.8 mL min-1, hexane:IPA (95:5),
20 ºC, tR 13.0 min (S) and tR 14.9 (R) min. Column IA.
[α]D25 = -13.3, c 0.5, CHCl3, for 90% ee of (R)-enantiomer.28
(S)-1-(4-Methoxyphenyl)ethanol, 7b
1H NMR (CDCl3, 300.13 MHz): d 1.48 (3H, d, J 6.3 Hz,
H2), 3.82 (3H, s, OCH3), 4.84 (1H, q, J 6.3 Hz, H1), 6.88
(2H, d, J 8.7 Hz, H3’ + H5’), 7.30 (2H, d, J 8.7 Hz, H2’ + H6’);
13C NMR (CDCl3, 75.5 MHz): d 24.3 (C2), 55.2 (OMe),
69.8 (C1), 126.6 (C2’ + C6’), 113.8 (C3’ + C5’), 137.9 (C1’),
158.9 (C4’). Conditions for determination of the conversion
by HPLC: 0.8 mL min-1, Hexane:IPA (95:5), 20 ºC,
tR (7a) 7.3 min and tR (7b) 8.5 min. Column Spherisorb.
Conditions for determination of the ee by HPLC:
0.8 mL min-1; hexane:IPA (95:5); 20 ºC, tR 13.4 min (S) and
tR 18.7 (R) min. Column OB-H. [α]D25 = -2.2, c 0.5, CH2Cl2,
for 9% ee of (S)-enantiomer.24
Supplementary Information
Supplementary data for 1b-10b alcohols are available
free of charge at http://jbcs.sbq.org.br, as a PDF file.
Acknowledgments
The authors thank to the Brazilian and Spanish agencies
CAPES-DGU (Process: 149/07), CNPq, FUNCAP,
PRONEX, Ministerio de Ciencia e Innovación (MICINN,
Project CTQ 2007-61126) for fellowships and financial
support. G. de G. and V. G.-F. thank MICINN for personal
grants (Juan de la Cierva and Ramon y Cajal program,
respectively).
(S)-1-(4-Chlorophenyl)ethanol, 8b
1H NMR (CDCl3, 300.13 MHz): d 1.45 (1H, d, J 6.5 Hz,
H1), 4.87 (q, J 6.5 Hz, H2), 7.30 (m, 4H, H2’ + H3’ +H5’ +
H6’); 13C NMR (CDCl3, 75.5 MHz): d 25.1 (C2), 69.6 (C1),
126.7 (C2’), 128.5 (C3’), 132.9 (C4’), 144.1 (C1’). Conditions
for determination of the conversion by GC: Injector 225 ºC,
Detector 250 ºC, 90 ºC (3 ºC min-1) 105 ºC (5 ºC min-1)
120 ºC (20 ºC min-1) 200 ºC (0 min), tR (8a) 5.0 min and tR
(8b) 5.6 min. Column HP-1. Conditions for determination
of the ee by HPLC: 0.8 mL min-1, hexane:IPA (95:5),
20 ºC, tR 7. min (S) and tR 8.5 min (R). Column OB-H.
[α]D25 = -32.4, CH2Cl2, c 0.5, for 62% ee of (S)-enantiomer.24
References
1. Kurbanoglu, E. B.; Zilbeyaz, K.; Kurbanoglu, N. I.; Kilic, H.;
Tetrahedron: Asymmetry 2007, 18, 2332.
2. Costa, L. F.A.; Lemos, F.; Ribeiro, F. R.; Cabral, J. M. S.; Catal.
Today 2008, 133-135, 625.
3. Andersson, P. G.; Munslow, I. J.; Modern Reduction Methods,
Wiley: Weinheim, 2008.
(R)-2-Chlorophenylethan-1-ol, 9b
1H NMR (CDCl3, 300.13 MHz): d 3.56 (1H, dd, J 11.4
and 3.3 Hz, H2), 3.66 (1H, dd, J 11.4 and 8.7 Hz, H2), 4.81
(1H, dd, J 8.7 and 3.6 Hz, H1), 7.30 (5H, m, H2’ + H3’ +
H4’ + H5’ + H6’); 13C NMR (CDCl3, 75.5 MHz): d 50.8 (C2),
74.0 (C1), 126.0 (C2’ + C6’), 128.4 and 128.6 (C3’ + C5’),
139.9 (C1’). Conditions for determination of the conversion
by GC: Injector 225 ºC, Detector 250 ºC, 90 ºC (3 ºC min-1)
105 ºC (5 ºC min-1) 120 (20 ºC min-1) 200 ºC (0 min), tR
(9a) 4.1 min and tR (9b) 4.3 min. Column HP-1. Conditions
for determination of the ee by HPLC: 0.8 mL min-1,
hexane:IPA (95:5), 20 ºC, tR 13.2 (R) and tR 17.2 (S) min.
Column OB-H. [α]D25 = -18.8, c 0.26, CH2Cl2, for 89% ee
of (R)-enantiomer.26
4. Dauβmann, T.; Rosen, T. C.; Dünkelmann, P.; Eng. Life Sci.
2006, 6, 125; Matsuda, T.; Yamanaka, R.; Nakamura, K.;
Tetrahedron: Asymmetry 2009, 20, 513.
5. Jurcek, O.; Wimmerová, M.; Wimmer, Z.; Coord. Chem. Rev.
2008, 252, 767; Borges, K. B.; Borges, W. S.; Durán-Patrón,
R.; Pupo, M. T.; Bonato, P. S.; Collado, I. G.; Tetrahedron:
Asymmetry 2009, 20, 385.
6. Patel, R. N.; Stereoselective Biocatalysis, Marcel Dekker: New
York, 2000.
7. Banerjee, C.; Soni, P.; Biotechnol. J. 2006, 1, 80.
8. Liria, C. W.; Romagna, C. D.; Rodovalho, N. N.; Marana, S.
R.; Miranda, M. T. M.; J. Braz. Chem. Soc. 2008, 19, 1574;
Zanotto, S. P.; Romano, I. P.; Lisboa, L. U. S.; Duviosin Jr., S.;

1516
Candida tropicalis CE017: a New Brazilian Enzymatic Source
J. Braz. Chem. Soc.
Martins, M. K.; Lima, F. A.; Silva, S. F.; Albuquerque, P. M.;
19. Andrade, L. H.; Comasseto, J. V.; Rodrigues, D. F.; Pellizari,
V. H.; Porto, A. L. M.; J. Mol. Catal. B: Enzym. 2005, 33, 73.
20. Csajági, C.; Szatzker, G.; Toke, E. R.; Ürge, L.; Darvas, F.;
Poppe, L.; Tetrahedron: Asymmetry 2008, 19, 237.
21. Seror, S.; Collin, J.; Fiaud, J. C.; Zouioueche, L. A.; J. Mol.
Catal. A: Chem. 2006, 256, 85.
J. Braz. Chem. Soc. 2009, 20, 1046.
9. Barros-Filho, B. A.; Nunes, F. M.; Oliveira, M. C. F.; Lemos,
T. L. G.; Mattos, M. C.; de Gonzalo, G.; Gotor-Fernández,
V.; Gotor, V.; J. Mol. Catal. B: Enzym., doi:10.1016/j.
molcatb.2010.01.023.
10. Barros-Filho, B. A.; Oliveira, M. C. F.; Lemos, T. L. G.;
Mattos, M. C.; de Gonzalo, G.; Gotor-Fernández, V.; Gotor,
V.; Tetrahedron: Asymmetry 2009, 20, 1057.
22. Ma,Y.; Liu, H.; Chen, L.; Zhu, J.; Deng, J.; Org. Lett. 2003, 5,
2103.
23. Yang, S. D.; Shi,Y.; Sun, Z. H.; Zhao,Y.; Liang,Y.; Tetrahedron:
Asymmetry 2006, 17, 1895.
11. Andrade, L. H.; Keppler,A. F.; Schoenlein-Crusius, I. H.; Porto,
A. L. M.; Comasseto, J. V.; J. Mol. Catal. B: Enzym. 2004, 31,
129.
24. Du, D.; Fang, T.; Xu, J.; Zhang, S.; Org. Lett. 2006, 8, 1327.
25. Morris, D. J.; Hayes, A. M.; Wills, M.; J. Org. Chem. 2006, 71,
7035.
12. Almeida,A. C. S.;Araújo, L.A.; Costa,A. M.;Abreu, C.A. M.;
Lima, M. A. G. A.; Palha, M. L. A. P. F.; Eletron. J. Biotechnol.
2005, 8, 55.
26. Tanis, S. P.; Evans, B. R.; Nieman, J. A.; Parker, T. T.; Taylor,
W. D.; Heasley, S. E.; Herrington, P. M.; Perrault, W. R.; Hohler,
R. A.; Dolak, L. A.; Hester, M. R.; Seest, E. P.; Tetrahedron:
Asymmetry 2006, 17, 2154.
13. Silverman, R. B.; The Organic Chemistry of Drug Design and
Drug Action, Academic Press: San Diego, 1992; Barret, G. M.;
Graboski, G. G.; Chem. Rev. 1986, 86, 751.
27. Alcohol 10b is given as example of the numerical locants used
14. Trost, B. M.; Fleming, I.; Comprehensive Organic Synthesis,
Pergamon Press: NewYork, 1991, vol. 8; Ram, S.; Ehrenkaufer,
R. E.; Tetrahedron Lett. 1984, 25, 3415;Yuste, F.; Saldana, M.;
Walls, F.; Tetrahedron Lett. 1982, 23, 147.
for NMR assignment:
OH
6'
Cl
1'
5'
1
2
2'
4'
Cl
15. Mangas-Sánchez, J.; Rodrígez-Mata, M.; Busto, E.; Gotor-
Fernández, V.; Gotor, V.; J. Org. Chem. 2009, 15, 5304.
16. Barbieri, C.; Bossi, L.; D’arrigo, P.; Fantoni, G. P.; Servi, S.;
J. Mol. Catal. B: Enzym. 2001, 11, 415.
3'
Cl
28. Zhu, D.; Mukherjee, C.; Hua, L.; Tetrahedron:Asymmetry 2005,
16, 3275.
17. Rocha, L. L.; Cordeiro, R. A.; Cavalcante, R. M.; Nascimento,
R. F.; Martins, S. C. S.; Santaella, S. T.; Melo, V. M. M.;
Mycopathologia 2007, 164, 183.
Received: December 18, 2009
Web Release Date: April 15, 2010
18. de Hoog, G. S.; Guarro, J.; Gene, J.; Figueras, M. J.; Atlas
of Clinical Fungi, 2nd ed., Réus: Centraalbureau voor
Schimmelcultures/Universitat Rovira i Virgili: Spain, 2000.

J. Braz. Chem. Soc., Vol. 21, No. 8, S1-S21, 2010.
Printed in Brazil - ©2010 Sociedade Brasileira de Química
0103 - 5053 $6.00+0.00
Candida tropicalis CE017: A New Brazilian Enzymatic Source for the Bioreduction
of Aromatic Prochiral Ketones
Gizelle A. B. Vieira,a Daniel M. de Freitas Araujo,a Telma L. G. Lemos,a
Marcos Carlos de Mattos,*,a Maria da Conceição F. de Oliveira,a Vânia M. M. Melo,a
Gonzalo de Gonzalo,b Vicente Gotor-Fernándezb and Vicente Gotor*,b
aDepartamento de Química Orgânica e Inorgânica, Universidade Federal do Ceará,
60451-970 Fortaleza-CE, Brazil
bDepartamento de Química Orgánica e Inorgánica, Instituto Universitario de Biotecnología de
Asturias, Universidad de Oviedo, c/ Julián Clavería 8, 33006, Oviedo, Spain
(S)-1-Phenylethanol, 1b
13C NMR (CDCl3, 75.5 MHz): d 25.3 (C2), 70.5 (C1),
125.5 (C2’ +C6’), 127.6 (C4’), 128.6 (C3’ + C5’), 146.0 (C1’).
(Figure S2)
OH
6'
1'
Conditions for determination of the conversion by
HPLC: 0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR (1a)
5.5 in and tR (1b) 8.5 min. Column Spherisorb. (Figure S3)
Conditions for determination of the ee by HPLC:
0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR 10.2 (S) and
tR 15.4 (R) min. Column OB-H. [α]D25 = -47.0, c 1.1, CHCl3,
for 97% ee of (S)-enantiomer. lit.: [α]D22 = -62.8 (c 1.0,
CHCl3), 98.5% ee. (Figures S4 and S5)
5'
1
2
2'
4'
3'
1H NMR (CDCl3, 300.13 MHz): d 1.39 (d, J 6.5 Hz,
3H, H2), 2.03 (s, 1H, OH), 4.77 (q, J 6.5 Hz, 1H, H1), 7.25
(m, 5H, H2’ + H3’ + H4’ + H5’+ H6’). (Figure S1)
Figure S1. 1H NMR spectrum of 1b (CDCl3, 300.13 MHz).
*e-mail: mcdmatto@ufc.br; vgs@fq.uniovi.es

S2
Candida tropicalis CE017: A New Brazilian Enzymatic Source
J. Braz. Chem. Soc.
Figure S2. 13C NMR spectrum of 1b (CDCl3, 75.5 MHz).
Figure S3. Chromatogram of the separation: tR (1a) 5.5 min and tR (1b) 8.5 min.
Figure S4. Chromatogram of the racemic mixture of 1b: tR 10.2 (S) and tR 15.4 (R) min.

Vol. 21, No. 8, 2010
Vieira et al.
S3
Figure S5. Chromatogram of the product by C. tropicalis (1b).
(S)-1-(2-Nitrophenyl)ethanol, 2b
13C NMR (CDCl3, 75.5 MHz): d 24.2 (C2), 65.4 (C1),
124.2 (C3’), 127.5 (C6’), 128.0 (C4’), 133.5 (C5’), 140.9
(C1’ + C2’). (Figure S7)
Conditions for determination of the conversion by GC:
Injector 225 oC, Detector 250 oC, 80 oC (5 oC min-1) 180 oC
(0 min), tR (2a) 8.4 min and tR (2b) 9.2 min. Column HP-1.
(Figure S8)
OH
6'
1'
5'
1
2
2'
4'
NO2
3'
Conditions for determination of the ee by HPLC:
0.8 mL min-1, hexane:IPA (97:3), 20 ºC, tR 22.1 min (R) and
tR 23.9 (S) min. Column AS. [α]D25 = +29.8, c 0.4, CHCl3,
for 99% ee of (S)-enantiomer. (Figures S9 and S10)
1H NMR (CDCl3, 300.13 MHz): d 1.55 (3H, d, J 6.3 Hz,
H2), 5.40 (1H, q, J 6.3 Hz, H1), 7.41 (1H, dt, J 8.1 and 1.2 Hz,
H4’),7.63(1H,dt,J8.1and1.5Hz,H5’),7.82(1H,dd,J8.1and
1.2 Hz, H6’), 7.88 (1H, dd, J 8.1 and 1.5 Hz, H3’). (Figure S6)
Figure S6. 1H NMR spectrum of 2b (CDCl3, 300.13 MHz).

S4
Candida tropicalis CE017: A New Brazilian Enzymatic Source
J. Braz. Chem. Soc.
Figure S7. 13C NMR spectrum of 2b (CDCl3, 75.5 MHz).
Figure S8. Chromatogram of the separation: tR (2a) 8.4 min and tR (2b) 9.2 min.
Figure S9. Chromatogram of the racemic mixture of 2b: tR 22.1 min (R) and tR 23.9 (S) min.

Vol. 21, No. 8, 2010
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S5
Figure S10. Chromatogram of the product by C. tropicalis (2b).
(S)-1-(3-Nitrophenyl)ethanol, 3b
and 0.9 Hz, H4’), 8.18 (1H, t, J 1.8 Hz, H2’). (Figure S11)
13C NMR (CDCl3, 75.5 MHz): d 25.2 (C2), 69.1 (C1),
OH
6'
1'
120.2 (C4’), 122.1 (C2’), 129.3 (C5’), 131.6 (C6’), 147.8 (C1’),
148.1 (C3’). (Figure S12)
5'
1
Conditions for determination of the conversion by GC:
Injector 225 oC, Detector 250 oC, 80 oC (5 oC min-1) 180 oC
(0 min), tR (3a) 9.5 min and tR (3b) 11.4 min. Column HP-
1. (Figure S13)
2
2'
4'
3'
NO2
Conditions for determination of the ee by HPLC:
0.8 mL min-1, hexane:IPA (90:10), 20 ºC, tR 12.8 min (S) and
tR 14.2 (R) min. Column OB-H. [α]D25 = -34.9, c 0.5 CH2Cl2,
for 99% ee of (S)-enantiomer. (Figures S14 and S15)
1H NMR (CDCl3, 300.13 MHz): d 1.47 (3H, d, J 6.6
Hz, H2), 4.96 (1H, q, J 6.6 Hz, H1), 7.47 (1H, t, J 7.8 Hz,
H5’), 7.67 (1H, d, J 7.8 Hz, H6’), 8.04 (1H, ddd, J 8.3, 2.1
Figure S11. 1H NMR spectrum of 3b (CDCl3, 300.13 MHz).

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Candida tropicalis CE017: A New Brazilian Enzymatic Source
J. Braz. Chem. Soc.
Figure S12. 13C NMR spectrum of 3b (CDCl3, 75.5 MHz).
Figure S13. Chromatogram of the separation: tR (3a) 9.5 min and tR (3b) 11.4 min.
Figure S14. Chromatogram of the racemic mixture of 3b: tR 12.8 min (S) and tR 14.2 (R) min.

Vol. 21, No. 8, 2010
Vieira et al.
S7
Figure S15. Chromatogram of the product by C. tropicalis (3b).
(S)-1-(4-Nitrophenyl)ethanol, 4b
13C NMR (CDCl3, 75.5 MHz): d 25.3 (C2), 69.5 (C1),
OH
123.6 (C2’ + C6’), 126.0 (C3’ + C5’), 147.0 (C1’), 153.1 (C4’).
(Figure S17)
6'
1'
Conditions for determination of the conversion by GC:
Injector 225 oC, Detector 250 oC, 80 oC (5 oC min-1) 180 oC
(0 min), tR (4a) 9.6 min and tR (4b) 11.8 min. Column
HP-1. (Figure S18)
5'
1
2
2'
4'
O2N
3'
Conditions for determination of the ee by HPLC:
0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR 11.7 min (R) and
tR 13.6 (S) min. Column AS. [α]D25 = -24.6, c 0.5, CHCl3,
for 96% ee of (S)-enantiomer. (Figures S19 and S20)
1H NMR (CDCl3, 300.13 MHz): d 1.50 (3H, d, J 6.6 Hz,
H2), 5.01 (1H, q, J 6.6 Hz, H1), 7.53 (2H, d, J 8.7 Hz, H2’ +
H6’), 8.17 (2H, d, J 8.7 Hz, H3’ + H5’). (Figure S16)
Figure S16. 1H NMR spectrum of 4b (CDCl3, 300.13 MHz).

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Candida tropicalis CE017: A New Brazilian Enzymatic Source
J. Braz. Chem. Soc.
Figure S17. 13C NMR spectrum of 4b (CDCl3, 75.5 MHz).
Figure S18. Chromatogram of the separation: tR (4a) 9.6 min and tR (4b) 11.8 min.
Figure S19. Chromatogram of the racemic mixture of 4b: tR 11.7 min (R) and tR 13.6 (S). min.

Vol. 21, No. 8, 2010
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S9
Figure S20. Chromatogram of the product by C. tropicalis (4b).
(S)-1-(2-Methoxyphenyl)ethanol, 5b
Hz, H5’), 7.29 (1H, dt, J 8.5 and 1.5 Hz, H6’), 7.36 (1H, dd,
J 7.5 and 1.5 Hz, H4’). (Figure S21)
13C NMR (CDCl3, 75.5 MHz): d 22.8 (C2), 55.2 (OCH3),
66.3 (C1), 110.3 (C3’), 120.7 (C5’), 126.0 (C6’), 128.2 (C4’),
133.4 (C1’), 156.4 (C2’). (Figure S22)
ConditionsfordeterminationoftheconversionbyHPLC:
0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR (5a) 6.3 min and
tR (5b) 7.1 min. Column Spherisorb. (Figure S23)
Conditions for determination of the ee by HPLC:
0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR 10.3 (S) and tR
16.8 (R) min. Column OB-H. [α]D25 = -4.5, c 0.4, CH2Cl2,
for 56% ee of (S)-enantiomer. (Figures S24 and S25)
OH
6'
1'
5'
1
2
2'
4'
OMe
3'
1H NMR (CDCl3, 300.13 MHz): d 1.52 (3H, d, J 6.6 Hz,
H2), 3.88 (3H, s, OCH3), 5.11 (1H, q, J 6.6 Hz, H1), 6.90
(1H, dd, J 7.5 and 1.5 Hz, H3’), 6.98 (1H, dt, J 8.5 and 1.5
Figure S21. 1H NMR spectrum of 5b (CDCl3, 300.13 MHz).

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Candida tropicalis CE017: A New Brazilian Enzymatic Source
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Figure S22. 13C NMR spectrum of 5b (CDCl3, 75.5 MHz).
Figure S23. Chromatogram of the separation: tR (5a) 6.3 min and tR (5b).
Figure S24. Chromatogram of the racemic mixture of 5b: tR 10.3 (S) and tR 16.8 (R) min.

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Figure S25. Chromatogram of the product by C. tropicalis (5b).
(S)-1-(3-Methoxyphenyl)ethanol, 6b
7.26 (1H, t, J 8.5 Hz, H5’). (Figure S26)
13C NMR (CDCl3, 75.5 MHz): d 25.0 (C2), 55.1 (OCH3),
OH
70.0 (C1), 110.8 (C2’), 112.7 (C4’), 117.6 (C6’), 129.3 (C5’),
147.6 (C1’), 159.6 (C3’). (Figure S27)
6'
1'
5'
1
ConditionsfordeterminationoftheconversionbyHPLC:
0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR (6a) 5.1 min and
tR (6b) 5.9 min. Column Spherisorb. (Figure S28)
Conditions for determination of the ee by HPLC:
0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR 20.1 min (S)
and tR 29.6 min (R). Column OB-H. [α]D25 = -21.6, c 0.34,
MeOH, for 80% ee of (S)-enantiomer. (Figures S29 and
S30)
2
2'
4'
3'
OMe
1H NMR (CDCl3, 300.13 MHz): d 1.47 (3H, d, J 6.6
Hz, H2), 3.81 (3H, s, OCH3), 4.83 (1H, q, J 6.6 Hz, H1),
6.81 (1H, dd, J 8.5 and 1.2 Hz, H6’), 6.93 (2H, m, H2’ + H4’),
Figure S26. 1H NMR spectrum of 6b (CDCl3, 300.13 MHz).

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Candida tropicalis CE017: A New Brazilian Enzymatic Source
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Figure S27. 13C NMR spectrum of 6b (CDCl3, 75.5 MHz).
Figure S28. Chromatogram of the separation: tR (6a) 5.1 min and tR (6b) 5.9 min.
Figure S29. Chromatogram of the racemic mixture of 6b: tR 20.1 min (S) and tR 29.6 min (R).

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Figure S30. Chromatogram of the product by C. tropicalis (6b).
(S)-1-(4-Methoxyphenyl)ethanol, 7b
13C NMR (CDCl3, 75.5 MHz): d 24.3 (C2), 55.2 (OMe),
69.8 (C1), 126.6 (C2’ + C6’), 113.8 (C3’ + C5’), 137.9 (C1’),
158.9 (C4’). (Figure S32)
OH
6'
1'
Conditions for determination of the conversion by
HPLC: 0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR (7a)
7.3minandtR (7b)8.5min. ColumnSpherisorb. (FigureS33)
Conditions for determination of the ee by HPLC:
0.8 mL min-1; hexane:IPA (95:5); 20 ºC, tR 13.4 min (S)
and tR 18.7 (R) min. Column OB-H. [α]D25 = -2.2, c 0.5,
CH2Cl2, for 9% ee of (S)-enantiomer. (Figures S34 and
S35)
5'
1
2
2'
4'
MeO
3'
1H NMR (CDCl3, 300.13 MHz): d 1.48 (3H, d, J 6.3
Hz, H2), 3.82 (3H, s, OCH3), 4.84 (1H, q, J 6.3 Hz, H1),
6.88 (2H, d, J 8.7 Hz, H3’ + H5’), 7.30 (2H, d, J 8.7 Hz, H2’
and H6’). (Figure S31)
Figure S31. 1H NMR spectrum of 7b (CDCl3, 300.13 MHz).

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Candida tropicalis CE017: A New Brazilian Enzymatic Source
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Figure S32. 13C NMR spectrum of 7b (CDCl3, 75.5 MHz).
Figure S33. Chromatogram of the separation: tR (7a) 7.3 min and tR (7b) 8.5 min.
Figure S34. Chromatogram of the racemic mixture of 7b: tR 13.4 min (S) and tR 18.7 (R) min.

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Figure S35. Chromatogram of the product by C. tropicalis (7b).
(S)-1-(4-Chlorophenyl)ethanol, 8b
13C NMR (CDCl3, 75.5 MHz): d 25.1 (C2), 69.6 (C1),
OH
126.7 (C2’), 128.5 (C3’), 132.9 (C4’), 144.1 (C1’). (Figure S37)
Conditions for determination of the conversion by GC:
Injector 225 ºC, Detector 250 ºC, 90 ºC (3 ºC min-1) 105 ºC
(5 ºC min-1) 120 ºC (20 ºC min-1) 200 ºC (0 min), tR (8a)
5.0 min and tR (8b) 5.6 min. Column HP-1. (Figure S38)
Conditions for determination of the ee by HPLC:
0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR 7.5 min
(S) and tR 8.5 min (R). Column OB-H. [α]D25 = -32.4,
CH2Cl2, c 0.5, for 62% ee of (S)-enantiomer. (Figures S39
and S40)
6'
1'
5'
1
2
2'
4'
Cl
3'
1H NMR (CDCl3, 300.13 MHz): d 1.46 (3H, d, J 6.5 Hz,
H1), 4.85 (1H, q, J 6.5 Hz, H2), 7.30 (4H, m, H2’+ H3’+ H5’ +
H6’). (Figure S36)
Figure S36. 1H NMR spectrum of 8b (CDCl3, 300.13 MHz).

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Candida tropicalis CE017: A New Brazilian Enzymatic Source
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Figure S37. 13C NMR spectrum of 8b (CDCl3, 75.5 MHz).
Figure S38. Chromatogram of the separation: tR (8a) 5.0 min and tR (8b) 5.6 min.
Figure S39. Chromatogram of the racemic mixture of 8b: tR 7.5 min (S) and tR 8.5 min (R).

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Figure S40. Chromatogram of the product by C. tropicalis (8b).
(R)-2-Chlorophenylethan-1-ol, 9b
13C NMR (CDCl3, 75.5 MHz): d 50.8 (C2), 74.0 (C1),
126.0 (C2’ + C6’), 128.4 and 128.6 (C3’ + C5’), 139.9 (C1’).
(Figure S42)
OH
6'
Cl
1'
Conditions for determination of the conversion by GC:
Injector 225 ºC, Detector 250 ºC, 90 ºC (3 ºC min-1) 105 ºC
(5 ºC min-1) 120 (20 ºC min-1) 200 ºC (0 min), tR (9a) 4.1 min
and tR (9b) 4.3 min. Column HP-1. (Figure S43)
Conditions for determination of the ee by HPLC:
0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR 13.2 (R) and tR
17.6 (S) min. Column OB-H. [α]D25 = -18.8, c 0.26, CH2Cl2,
for 89% ee of (R)-enantiomer. (Figures S44 and S45)
5'
1
2
4'
2'
3'
1H NMR (CDCl3, 300.13 MHz): d 3.56 (1H, dd, J 11.4
and 3.3 Hz, H2), 3.66 (1H, dd, J 11.4 and 8.7 Hz, H2), 4.81
(1H, dd, J 8.7 and 3.6 Hz, H1), 7.30 (5H, m, H2’ + H3’ + H4’ +
H5’ +H6’). (Figure S41)
Figure S41. 1H NMR spectrum of 9b (CDCl3, 300.13 MHz).

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Candida tropicalis CE017: A New Brazilian Enzymatic Source
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Figure S42. 13C NMR spectrum of 9b (CDCl3, 75.5 MHz).
Figure S43. Chromatogram of the separation: tR (9a) 4.1 min and tR (9b) 4.3 min.
Figure S44. Chromatogram of the racemic mixture of 9b: tR 13.2 (R) and tR 17.6 (S) min.

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Figure S45. Chromatogram of the product by C. tropicalis (9b).
(R)-2-Chloro-1-(3,4-dichlorophenyl)ethan-1-ol, 10b
1.8 Hz, H6’), 7.43 (1H, d, J 8.4 Hz, H5’), 7.51 (1H, d, J 1.8
Hz, H2’). (Figure S46)
OH
13C NMR (CDCl3, 75.5 MHz): d 50.4 (C2), 72.8 (C1),
125.4 (C6’), 128.1 (C2’), 130.6 (C5’), 132.4 (C4’), 132.8 (C3’),
140.0 (C1’). (Figure S47)
6'
Cl
1'
5'
1
2
Conditions for determination of the conversion by GC:
Injector 225 ºC, Detector 250 ºC, 80 ºC (5 ºC min-1) 180 ºC
(0 min); tR (10a) 13.6 and tR (10b) 14.8 min. Column HP-1.
(Figure S48)
Conditions for determination of the ee by HPLC:
0.8 mL min-1, hexane:IPA (95:5), 20 ºC, tR 13.0 min (S) and
tR 14.9 (R) min. Column IA. [α]D25 = -13.3, c 0.5, CHCl3,
for 90% ee of (R)-enantiomer. (Figures S49 and S50)
2'
4'
Cl
3'
Cl
1H NMR (CDCl3, 300.13 MHz): d 3.59 (1H, dd, J 11.4
and 8.7 Hz, H2), 3.72 (1H, dd, J 11.4 and 3.6 Hz, H2), 4.87
(1H, dd, J 8.7 and 3.6 Hz, H1), 7.21 (1H, dd, J 8.4 and
Figure S46. 1H NMR spectrum of 10b (CDCl3, 300.13 MHz).

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Candida tropicalis CE017: A New Brazilian Enzymatic Source
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Figure S47. 13C NMR spectrum of 10b (CDCl3, 75.5 MHz).
Figure S48. Chromatogram of the separation: tR (10a) 13.6 and tR (10b) 14.8 min.
Figure S49. Chromatogram of the racemic mixture of 10b: tR 13.0 min (S) and tR 14.9 (R) min.

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Figure S50. Chromatogram of the product by C. tropicalis (10b).