Bioreduction of acetophenone derivatives by red marine algae Bostrychia radicans and B. tenella, and marine bacteria associated

Article abstract of DOI:10.1002/hlca.201000434

The biocatalytic reduction of acetophenone derivatives was exploited by using algal biomass from Bostrychia radicans and B. tenella producing exclusively (S)-2-phenylethanols with high enantiomeric excess (>99% ee). Bacterial populations associated with algal biomass were identified as the Bacillus genus. This report deals with the first investigations involving the use of marine bacteria associated with B. radicans and B. tenella marine algae for the biocatalytic reduction of acetophenone derivatives. Copyright

Full text of DOI:10.1002/hlca.201000434

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Helvetica Chimica Acta – Vol. 94 (2011)
Bioreduction of Acetophenone Derivatives by Red Marine Algae Bostrychia
radicans and B. tenella, and Marine Bacteria Associated
a
a
b
b
by Ana M. Mouad ), Mariana P. Martins ), Hosana M. Debonsi ), Ana L. L. de Oliveira ),
b
c
c
d
Rafael de Felicio ), Nair S. Yokoya ), Mutue T. Fujii ), Cla u´ dia B. A. de Menezes ),
d
a
Fabiana Fantinatti-Garboggini ), and Andr e´ L. M. Porto* )
a) Instituto de Quꢀmica de S a˜ o Carlos, Universidade de S a˜ o Paulo, Av. Trabalhador S a˜ o-carlense,
4
00, CEP 13560-970, CP 780, S a˜ o Carlos-SP, Brazil
(
phone: þ 551633738103; fax: þ 551633739952; e-mail: almporto@iqsc.usp.br)
b) Faculdade de Ci eˆ ncias Farmac eˆ uticas de Ribeir a˜ o Preto, Departamento de Fꢀsica e Quꢀmica,
Universidade de S a˜ o Paulo, Via do Caf e´ s/n, CEP 14040-903, Ribeir a˜ o Preto-SP, Brazil
c) Instituto de Bot aˆ nica de S a˜ o Paulo, Av. Miguel Est e´ fano, 3687, CEP 04301-902, CP 4005,
´
Agua Funda, S a˜ o Paulo-SP, Brazil
d) Recursos de Divis a˜ o Microbianos, Centro Pluridisciplinar de Pesquisas Quꢀmicas, Biol o´ gicas e
Agrꢀcolas, CPQBA, Universidade Estadual de Campinas, Rua Alexandre Casellato, 999,
CEP 13140-000, Paulꢀnia-SP, Brazil
The biocatalytic reduction of acetophenone derivatives was exploited by using algal biomass from
Bostrychia radicans and B. tenella producing exclusively (S)-2-phenylethanols with high enantiomeric
excess (> 99% ee). Bacterial populations associated with algal biomass were identified as the Bacillus
genus. This report deals with the first investigations involving the use of marine bacteria associated with
B. radicans and B. tenella marine algae for the biocatalytic reduction of acetophenone derivatives.
1. Introduction. – Reduction reactions of prochiral ketones promote the production
of enantiomerically pure alcohols which are valuable intermediates in asymmetric
organic synthesis. Microorganisms from different natural sources may carry out these
biotransformations. Marine biodiversity has an enormous potential for biotechnolog-
ical processes. Marine metagenomics offers strategies for the discovery of new enzymes
with biotechnological applications [1][2].
The biocatalytic reduction of prochiral ketones has been extensively studied by
several methods [3]. However, the use of algae for the reduction of ketones is rare in
comparison to cells of microorganisms, such as fungi, yeast, and bacteria [4][5].
Yoshizako et al. [6 – 9] described the asymmetric reduction of the CO groups using the
Chorella pyrenoidosa alga. Biotransformation of aliphatic and aromatic ketones, and
aldehydes were reported from five cultures of photosynthetic marine microalgae
[
10][11]. Dunaliella tertiolecta (green microalga) catalyzed the biotransformation of
terpenoids [12], and several keto esters were converted to the corresponding hydroxy
esters by marine microalgae such Chaetoceros and Nannochloropsis [13]. The Spirulina
platensis alga catalyzed alkanone debromation with good yields [14]. There are
excellent studies involving the reduction of various ketones by red algae yielding
enantiomerically pure alcohols [4].
ꢁ
2011 Verlag Helvetica Chimica Acta AG, Zꢂrich

Helvetica Chimica Acta – Vol. 94 (2011)
1507
Recently, we reported the bioreduction of a-chloroacetophenone and the
biotransformation of a-bromoacetophenones using marine fungi Trichoderma sp.,
Penicillium mickyzinskii, and Bionectria sp. [15][16]. These fungi catalyzed these
biotransformations only when they were growing in artificial seawater.
Here, we present the use of marine algae Bostrychia tenella and B. radicans, and
their associated microorganisms for the reduction of acetophenone derivatives. In
addition, we isolated strains of marine bacteria of the Bacillus genus from B. radicans
which catalyzed the enantioselective reduction of acetophenone derivatives with high
enantiomeric excess (> 99% ee).
2. Experimental. – 2.1. General. All manipulations involving algae and marine bacteria were carried
out under sterile conditions in a Veco laminar flow cabinet. Technal TE-421 or Superohm G-25 orbital
shakers were employed in the biocatalytic transformation experiments. Reagents ortho-iodoacetophe-
none (1), meta-iodoacetophenone (2), para-iodoacetophenone (3), ortho-fluoroacetophenone (4),
ortho-chloroacetophenone (5), ortho-bromoacetophenone (6) and ortho-nitroacetophenone (7), and
NaBH were purchased from SigmaꢀAldrich. Products derived synthetically or from enzymatic processes
4
were purified by column chromatography (CC) on silica gel (SiO ; 230 – 400 mesh) eluted with hexane/
2
AcOEt mixtures. The column eluent was monitored by TLC on pre-coated silica gel 60 F254 layers
(
aluminum-backed: Sorbent) eluted with hexane/AcOEt 9 :1 and 8 :2. Reaction products were analyzed
using a Shimadzu model 2010 GC/FID equipped with an auto-injector AOC20i and Varian CP-Chiralsil-
DEX b-Cyclodextrin column (25 m ꢁ 0.25 mm i.d.; 0.39 mm). The programs used for GC analyses of
alcohols are described in Table 1. The injector and detector were maintained at 2008, the split ratio of the
injector was 1:20, and the carrier gas was N at 60 kPa. The ee values of alcohols were determined by GC
2
analyses.
Table 1. Programs Used for Identification of the Alcohols 8 – 14 by GC Analysesa)
Alcohols
T [8]
i
ti [min]
T [8]
f
tf [min]
r [8/min]
tR [min]
8
9
115
120
120
100
60
2
2
2
2
2
2
2
175
190
165
180
180
140
150
8
8
8
5
5
6
6
3
2
2
2
2
2
2
(R) 18.0, (S) 20.0
(R) 21.0, (S) 22.5
(R) 21.0, (S) 22.0
(R) 21.0, (S) 22.0
(R) 23.0, (S) 25.0
(R) 24.0, (S) 26.0
(R) 26.0, (S) 27.0
1
1
1
1
1
0
1
2
3
4
90
100
a) Chiral column: CP-Chiralsil-DEX b-Cyclodextrin (25 m ꢁ 0.25 mm i.d.; 0.39 mm); T : initial temper-
i
ature; T : final temperature; t : initial time; t : final time; r: rate; t : retention time.
f
i
f
R
2
.2. Synthesis of Racemic Alcohols 8 – 14. Racemic alcohols 8 – 14 were obtained by reduction of the
1
corresponding ketones 1 – 7 with NaBH and MeOH [17]. The spectroscopic data (IR, H-NMR, and
4
MS) of 8 – 14 were in agreement with those reported in the literature [18 – 22].
2.3. Collection and Identification of Algae. The algae Bostrychia tenella and Bostrychia radicans were
collected in Ubatuba, in the South Atlantic Ocean off the northern coast of the State of S a˜ o Paulo, Brazil,
in September 2007. Algae were identified by using conventional taxonomic methods by N. S. Yokoya
from the Instituto de Botanica de S a˜ o Paulo, Brazil [23]. Algal material (2 kg) was stored in a freezer
(
ꢀ148). Voucher specimens were deposited with the Herbarium of the Instituto de Bot aˆ nica (S a˜ o Paulo,
SP, Brazil) under the accession Nos. SP 365678 (B. radicans) and SP 400217 (B. tenella).
.4. Isolation and Identification of Marine Bacteria from B. tenella and B. radicans. Algae B. tenella
and B. radicans stored in a freezer at ꢀ 148 were transferred to a refrigerator at a temp. of 48, and then
2

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Helvetica Chimica Acta – Vol. 94 (2011)
immersed in dist. H O. The algae were washed with artificial seawater (pH 8), immersed for 30 s in
2
HClO (0.01%), and washed with dist. H O. The algal biomass (5.0 g) was transferred to 250-ml
2
Erlenmeyer flasks, containing 100 ml of H O (pH 8), and incubated at 328 for 5 d on a rotary shaker
2
(
150 rpm). Then, the algal culture was inoculated in Petri dishes containing nutrient broth. Triplicate
plates were incubated in an oven at 328 for 8 d. The microorganisms grown on the plates containing agar
nutrient broth were isolated and identified by 16S rRNA gene-sequence analysis.
Genomic DNA was obtained according to the method of Pospiech and Neumann [24]. 16S rRNA
gene was amplified by PCR (¼ polymerase chain reaction) using specific primers, 27f and 1401r for
universal Bacteria Domain [25].
Fifty-ml reaction mixtures contained 100 ng of total DNA, 2 U of Taq polymerase (Invitrogen),
0
.2 mm of deoxynucleoside triphosphates, and 0.4 mm of each primer. The PCR amplifications were
performed using an initial denaturation step of 2 min at 958, followed by 30 cycles of 1 min at 948, 1 min
at 558 and 3 min at 728, and the extension of 3 min at 728, in an Eppendorf thermal cycler.
TM
The PCR product was purified on GFX
PCR DNA and a Gel Band Purification kit (GE
HealthCare) for automated sequencing in the MegaBace DNA Analysis System 1000. The sequencing was
carried out using the 10f (5’GAG TTT GAT CCT GGC TCA G3’), 765f (5’ATT AGA TAC CCT GGT
AG3’), 782r (5’ACC AGG GTA TCT AAT CCT GT3’), and 1100r (5’AGG GTT GGG GTG GTT G 3’)
primers, and the DYEnamic ET Dye Terminator Cycle Sequencing Kit for the automated MegaBace 500
system (GE HealthCare), according to the manufacturerꢃs recommendations.
Partial 16S rRNA sequences obtained from isolates were assembled in a contig using the phred/
Phrap/CONSED program [26][27]. Identification was achieved by comparing the contiguous 16S rRNA
sequences obtained with 16S rRNA sequence data from reference and type strains available in the public
databases GenBank and RDP (Ribosomal Database Project, Michigan State University, USA) using the
BLASTn and Seqmatch, resp. The sequences were aligned using the CLUSTAL X program and analyzed
with MEGA v.4 software [28][29]. Evolutionary distances were derived from sequence-pair dissim-
ilarities calculated as implemented in MEGA, using Kimuraꢃs DNA substitution model [30]. The
phylogenetic reconstruction was achieved using the neighbor-joining (NJ) algorithm, with bootstrap
values calculated from 1000 replicate runs, using the routines included in the MEGA software [29][31].
Phylogenetic analysis of partial 16S rRNA gene sequences of bacterial associated from B. tenella and B.
radicans algae is shown in the Figure.
2.5. Biocatalytic Reduction of Iodoacetophenones 1 – 3 using B. tenella and B. radicans. The B. tenella
and B. radicans algae stored in the freezer at ꢀ 148 were transferred to a refrigerator at a temp. of 48, and
Figure. Phylogenetic analysis of partial 16S rRNA gene sequences of bacteria associated with marine algae
B. tenella and B. radicans. Evolutionary distances were based on Kimura 2p model and tree
reconstruction on the neighbor joining method. Bootstrap values (1000 replicate runs, shown as %)
>
70% are listed. Pseudomnas aeruginosa PAO1 (NC_002516.2) was used as outgroup.

Helvetica Chimica Acta – Vol. 94 (2011)
1509
then immersed in dist. H O. The algae were washed with artificial seawater (pH 8), immersed for 30 s in
2
HClO (0.01%), and washed with dist. H O. Algae (5.0 g) were transferred to 250-ml Erlenmeyer flasks
2
containing dist. H O (100 ml), and the ketones 1 – 3 (50.0 mg; 0.20 mmol) dissolved in 300 ml of DMSO
2
were added immediately. The Erlenmeyer flasks were shaken at 328 for 8 d on an orbital shaker
(
150 rpm). Afterwards, the algal biomass was harvested by filtration, and the aq. phases were extracted
with AcOEt (3 ꢁ 30 ml). The org. phases were separated, dried (Na SO ), and evaporated until dryness
2
4
under vacuum. The reactions products were analyzed by GC-FID, and the results are compiled in Table 2.
.6. Biocatalytic Reduction of ortho-Acetophenone Derivatives with Bacteria Associated from B.
2
tenella and B. radicans. The bacteria associated from B. tenella and B. radicans algae were used for the
reduction of ortho-acetophenones 4 – 7. The strains were preserved in agar culture containing nutrient
broth at 48. In addition, the strains were grown in liquid culture medium and incubated in 250-ml
Erlenmeyer flasks with 100 ml of nutrient broth at 328 for 48 h on an orbital shaker (150 rmp). Then, the
ortho-acetophenones (50.0 mg) dissolved in 300 ml of DMSO were added immediately. The Erlenmeyer
flasks were shaken at 328 for 4 – 8 d on an orbital shaker (150 rmp). The mixtures were extracted with
AcOEt (3 ꢁ 30 ml), and org. phases were separated, dried (Na SO ), and evaporated until dryness under
2
4
vacuum. The results are collected in Tables 3 – 5.
.7. Assignment of the Absolute Configurations. The optical rotations of compounds obtained from
2
the biocatalytic reactions were determined in 1-dm cuvette using a PerkinꢀElmer model 241 polarimeter
and were referenced to the Na-D line. The abs. configurations of alcohols 8 – 14 were determined by
comparing the specific signs of rotation measured for the products with those reported in the literature
(
Table 6).
3
. Results and Discussion. – Initially, the biocatalytic reductions of the iodoace-
tophenones 1 – 3 were carried out using red algae Bostrychia tenella and Bostrychia
radicans. The algal biomass B. tenella and B. radicans stored in a freezer at ꢀ 148 were
transferred to a refrigerator at a temperature of 48, and then immersed in distilled H O.
2
The algae were washed with artificial seawater (pH 8), immersed for 30 s in HClO
(
0.01%), and washed with distilled H O. The algae were used for reduction of ketones
2
1
– 3, and the results are summarized in Table 2.
One of the prime requirements of a biocatalytic reduction is the optimization of the
reaction parameters, i.e., amount of cell biomass, type of substrates, the effects of
reaction temperature, and time. In this study, we used ortho-, meta- and para-
iodoacetophenones 1 – 3. With the optimizations of the experiments, the reactions were
carried out for 8 d and then analyzed by GC. B. tenella and B. radicans algae yielded
enantiomerically pure (iodophenyl)ethanols 8 – 10 with high enantiomeric excesses
(> 99% ee). The absolute configurations of the alcohols 8 – 10 were in accordance with
Prelogꢃs rule, i.e., (S)-alcohols (Table 2).
In this study, we observed that the ortho-iodoacetophenone (1) afforded the best
results for the bioreduction by B. tenella and B. radicans algae. Concentrations of (S)-
alcohol 8 were obtained with 40 – 42% after 8 d of incubation (Table 2). However, the
reduction of meta-iodoacetophenone (2) and para-iodoacetophenone (3) with the two
algae produced lower amounts of the alcohols 9 and 10, respectively. Ketone 1 was
more reactive than compounds 2 and 3 under these conditions.
To test the capacity of the new biocatalyst, the biomass of B. tenella was used for the
reduction of the ketones 4 – 7 with different groups in the ortho-position. The different
groups strongly influenced the results (Table 3). The reduction of ortho-fluoroaceto-
phenone 4 yielded alcohol 11 with high enantiomeric excesses (> 99% ee) and 33%
concentration after 4 d of incubation (Table 3, Entry 1). Extending the reaction time to

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Helvetica Chimica Acta – Vol. 94 (2011)
Table 2. Bioreduction of the Iodoacetophenones 1 – 3 by Marine Algae Bostrychia tenella and B.
a
radicans )
c [%]b)
ee [%] ) )
c d
Entry
Algae
Ketones
Ketones
Alcohols
Alcohols (abs. conf.)
1
2
3
B. radicans
2-I 1
3-I 2
4-I 3
58
92
83
42e)
8
17
99 (S)
99 (S)
99 (S)
4
5
6
B. tenella
2-I 1
3-I 2
4-I 3
60
95
84
40
5
16
99 (S)
99 (S)
99 (S)
a) Reaction conditions: biomass (5.0 g) was transferred to 250-ml Erlenmeyer flasks containing distilled
H O (100 ml) with the ketones 1 – 3 (50.0 mg, 0.20 mmol) dissolved in 300 ml of DMSO. Erlenmeyer
2
b
flasks were shaken at 328 for 8 d on an orbital shaker (150 rmp). ) c [%]: Concentration determined by
c
d
GC. ) ee [%]: Enantiomeric excess. ) Chiral column: CP-Chiralsil-DEX b-Cyclodextrin (25 m ꢁ
e
0
.25 mm i.d.; 0.39 mm). ) Yield isolated: 25%.
more than 8 d, (S)-fluoroalcohol 11 was obtained with 52% concentration (Table 3,
Entry 2).
Surprisingly, the bulky groups in the ortho-position (Cl, Br, and NO ) led to a
2
decrease in the concentration of the alcohols 12 – 14 (Table 3), but high enantiomeric
excesses were obtained. Longer incubation times in these experiments led to higher
yields of alcohols 11 – 14 (Table 3). For example, the reduction of nitroacetophenone 7
did not occur after 4 d. The increase of the time to 8 d afforded alcohol 14 with 13%
concentration (Table 3, Entries 7 and 8).
During these studies employing biomass algae, we promoted the isolation of
associated microorganisms of the red algae, given that bacteria are opportunists and
capable of growth in algal culture [32]. The mild sterilization of algae (artificial
seawater and HClO) allowed the growth of the associated microorganisms. The B.
tenella and B. radicans algal cultures were cultivated on an orbital shaker containing the
distilled H O (see Experimental). In addition, the algal solutions were inoculated in
2
Petri dishes containing agar medium. The bacteria associated grown in agar medium
was transferred to liquid medium and the acetophenones dissolved in 300 ml of DMSO
were added (see Experimental).
Bacteria associated with algae promoted bioreduction of iodoacetophenones 1 – 3
with high enantiomeric excess (> 99% ee) with different concentrations (Table 4).
Strain BR62 from B. radicans catalyzed reductions of iodoacetophenones 1 and 3 with
46 and 48% concentrations, respectively, within 8 d (Table 4, Entries 2 and 6). Strain
BT01 from B. tenella under the same conditions afforded the alcohols 8 and 9 in good

Helvetica Chimica Acta – Vol. 94 (2011)
1511
Table 3. Bioreduction of the ortho-Acetophenones 4 – 7 by Marine Algae Bostrychia tenellaa)
c [%]b)
ee [%] ) )
d e
Entry
Ketones
Time [d]
Ketones
Alcohols (Yield [%])c)
Alcohols (abs. conf.)
1
2
3
4
5
6
7
8
2-I 4
4
8
4
8
4
8
4
8
67
48
93
80
95
90
100
87
33 (24)
52 (39)
99 (S)
95 (S)
99 (S)
98 (S)
99 (S)
99 (S)
0
2-Cl 5
2-Br 6
7
20
5
10
0
2-NO 7
2
13
99 (S)
a) Reaction conditions: biomass (5.0 g) was transferred to 250-ml Erlenmeyer flasks containing distilled
H O (100 ml) with the ketones 4 – 7 (50.0 mg) dissolved in 300 ml of DMSO. Erlenmeyer flasks were
2
b
c
shaken at 328 for 8 d on an orbital shaker (150 rmp). ) c [%]: Concentration determined by GC. ) Yield
isolated. ) ee (%): Enantiomeric excess. ) Chiral column: CP-Chiralsil-DEX b-Cyclodextrin (25 m ꢁ
.25 mm i.d.; 0.39 mm).
d
e
0
enantiomeric excess (> 99% ee), and low concentrations (Table 4, Entries 8 and 12).
meta-Iodoacetophenone 2 was reduced in poor concentrations and high enantiomeric
excess (Table 4).
Similarly, strain BR62 from B. tenella catalyzed the reduction of ortho-acetophe-
nones 4 – 7 (Table 5). Strain BR62 was also efficient in providing alcohols 11 – 13 with
high enantiomeric excesses and good concentrations (Table 5, Entries 1 – 3). In these
investigations, only the ortho-nitroacetophenone was obtained in poor optical purity
and concentration (Table 5, Entry 4).
The biocatalytic experiments demonstrated that marine algae B. tenella, B. radicans,
and associated bacteria catalyzed the bioreduction of acetophenone derivatives with
high optical purities. Strain BR62 associated from B. radicans was identified as Bacillus
sp., supporting values of 100% on the boot strap, similar to strain types B. pumilus and
B. safensis. The nucleotide alignment of strain BT01 supported bootstrap values of
99%, showing similarity to different Bacillus strains including the sequences of the
strains B. subtilis and B. licheniformis. In addition, the strains were identified as Bacillus
sp. through their morphology and by 16S rRNA gene sequence analysis (Fig.).
The bacterial cultures may be contaminants present in algal biomass [32]. The
associated bacteria with marine algae may be opportunists, utilizing algal extracellular
products. In the literature, the Bacillus strains were isolated from the brown marine
alga Sargassum sp. This report represents the first article to deal with Bacillus sp.

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Helvetica Chimica Acta – Vol. 94 (2011)
Table 4. Bioreduction of the Iodoacetophenones 1 – 3 by Bacteria Associated with Marine Algae
a
Bostrychia tenella and B. radicans )
c [%]b)
ee [%] ) )
d e
Entry
Ketones
Time [d]
Ketones
Alcohols (Yield [%])c)
Alcohols (abs. conf.)
Bacillus sp. strain BR62 associated with the marine alga B. radicans
1
2
3
4
5
6
2-I 1
3-I 2
4-I 3
4
8
4
8
4
8
97
54
98
83
96
52
3
99 (S)
99 (S)
99 (S)
99 (S)
99 (S)
99 (S)
46 (30)
2
17
4
48 (34)
Bacillus sp. strain BT01 associated with the marine alga B. tenella
7
8
9
0
1
2
2-I 1
3-I 2
4-I 3
4
8
4
8
4
8
94
70
92
87
91
80
6
99 (S)
99 (S)
99 (S)
99 (S)
99 (S)
99 (S)
30 (23)
8
13
9
1
1
1
20
a) Reaction conditions: bacterial consortiums of the Bacillus sp., isolated from marine algae B. radicans
and B. tenella were grown in Erlenmeyer flasks containing nutrient broth for 3 d, and then the ketones 1 –
3
(50.0 mg; 0.20 mmol) dissolved in 300 ml of DMSO were added. Erlenmeyer flasks were shaken at 328
for 8 d on an orbital shaker (150 rmp). ) c [%]: Concentration determined by GC. c) Yield isolated.
b
d
e
)
ee [%]: Enantiomeric excess. ) Chiral column: CP-Chiralsil-DEX b-Cyclodextrin (25 m ꢁ 0.25 mm
i.d.; 0.39 mm).
associated with interdital marine alga [33]. Recently, red algae species yielded marine
bacteria of the Bacillus genus [34].
4
. Conclusions. – In conclusion, the biocatalytic reduction of acetophenone
derivatives was accomplished using B. tenella and B. radicans algal biomass yielding
enantiomerically pure alcohols (> 99% ee). In addition, we isolated bacteria strains
associated with red marine algae B. tenella and B. radicans. Bacteria were identified as
Bacillus sp., and they catalyzed the bioreduction of the acetophenone derivatives with
high enantiomeric excess (> 99% ee). The use of the new marine organisms is
important for the discovery of new enzymes for biotechnological applications.
The authors wish to thank the Conselho Nacional de Desenvolvimento Cientꢀfico e Tecnol o´ gico
(
CNPq; grant No. 472663/2004-6 to A. L. M. P.) and the FundaÅ a˜ o de Amparo a` Pesquisa do Estado de
S a˜ o Paulo (FAPESP; grant No. 2006/54401-2 to A. L. M. P.) for their financial support. Thanks are also

Helvetica Chimica Acta – Vol. 94 (2011)
1513
Table 5. Bioreduction of the ortho-Acetophenones 4 – 7 by Bacillus sp. strain BR62 Isolated from Marine
a
Algae B. radicans )
d e
Entry
Ketones
c [%]b)
Ketones
ee [%] ) )
Alcohols (Yield [%])c)
Alcohols (abs. conf.)
1
2
3
4
2-F 4
2-Cl 5
2-Br 6
27
51
5
73 (59)
49 (35)
95 (77)
7
99 (S)
99 (S)
99 (S)
10 (S)
2-NO 7
93
2
a) Reaction conditions: bacteria isolated from marine alga B. radicans was grown in Erlenmeyer flasks
containing nutrient broth for 3 d, and then the ketones 4 – 7 (50.0 mg) dissolved in 300 ml of DMSO were
b
added. Erlenmeyer flasks were shaken at 328 for 8 d on an orbital shaker (150 rmp). ) c [%]:
Concentration determined by GC. ) Yield isolated. ) ee [%]: Enantiomeric excess. e) Chiral column:
c
d
CP-Chiralsil-DEX b-Cyclodextrin (25 m ꢁ 0.25 mm i.d.; 0.39 mm).
Table 6. Optical Rotations of the Alcohols 8 – 14
[a]2 (exper.)
5
[a]D (literature)
25
Alcohols
D
1
1
1
1
1
1
1
-(2-Iodophenyl)ethanol (8)
-(3-Iodophenyl)ethanol (9)
-(4-Iodophenyl)ethanol (10)
(S)-8 ꢀ 17.6 (c ¼ 1.7, CHCl ) (S)-8 ꢀ 41.3 (c ¼ 0.20, CHCl ) [17]
3
3
a
a
)
)
(S)-9 ꢀ 12.9 (c ¼ 2.4, CHCl
3
) [18]
(S)-10 ꢀ 32.7 (c ¼ 5.99, CHCl ) [18]
3
-(2-Fluorophenyl)ethanol (11) (S)-11 ꢀ 45.5 (c ¼ 2.0, CHCl ) (S)-11 ꢀ 34.7 (c ¼ 3.63, CH Cl ) [19]
3
2
2
a
-(2-Chlorophenyl)ethanol (12)
)
(S)-12 ꢀ 32.9 (c ¼ 0.22, CHCl ) [17]
3
-(2-Bromophenyl)ethanol (13) (R)-13 þ 26 (c ¼ 2.5, CHCl ) (S)-13 ꢀ 54.5 (c ¼ 0.20, CHCl ) [17]
3
3
-(2-Nitrophenyl)ethanol (14) (S)-14 þ 17 (c ¼ 1.8, CHCl ) (S)-14 þ 30.6 (c ¼ 3.6, CHCl ) [20]
3
3
a) Absolute configurations of 9, 10, and 12 were attributed by retention times on GC-FID chiral (see
Table 1) [19].
due to CoordenaÅ a˜ o de AperfeiÅoamento de Pessoal de Nꢀvel Superior (CAPES) and CNPq for
scholarships awarded to M. P.M. and A. M.M., respectively.
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Received November 28, 2010