SUPPORTING INFORMATION
Stereoselective Bioreduction of α-Azido Ketones by Whole Cells of Marinederived Fungi
Lenilson C. Rocha  Mirna H. R. Seleghim  João V. Comasseto  Lara D. Sette  André L.
M. Porto
Preparation of α-azido ketones 5-8
A mixture of the appropriate α-haloacetophenones 1-4 (14.0 mmol) and sodium azide (1.82
g, 28.0 mmol) in acetone (250 mL) was stirred at room temperature until completion of the
reaction and monitored by TLC. After that the mixture was poured into water and extracted
with EtOAc (3 x 30 mL), and the organic layer was dried with MgSO4, concentrated under
reduced pressure, and the residue was purified by column chromatography.
2-azido-1-phenylethanone (5): 1H NMR (200 MHz, CDCl3): δ 4.57 (s, 2H); 7.48-7.68 (m,
3H); 7.90-7.95 (m, 2H). IR (KBr,  = cm−1): 2103 (N3), 1695 (C=O).
7.68
7.66
7.64
7.63
7.61
7.61
7.60
7.55
7.55
7.52
7.51
7.48
7.95
7.94
7.91
7.90
002001.FID
2.15
3.39
7.75
Chemical Shift (ppm)
7.50
2.15
8.5
8.0
7.48
7.47
7.69
7.68
7.95
7.94
7.91
7.90
7.64
7.55
7.55
7.51
7.27
4.57
8.00
3.39
7.5
2.00
7.0
6.5
6.0
5.5
5.0
4.5
4.0
Chemical Shift (ppm)
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0
Figure S1. 1H NMR spectrum (200 MHz, CDCl3) of 2-azido-1-phenyletanone (5)
Wavenumber (cm-1)
Figure S2. IR spectrum (KBr) of 2-azido-1-phenylethanone (5)
-0.5
2-azido-1-(4-methoxyphenyl)ethanone (6): 1H NMR (200 MHz, CDCl3): δ 3.89 (s, 3H);
4.51 (s, 2H); 6.97 (d, 2H, J 8.0 Hz); 7.90 (d, 2H, J 8.0 Hz). IR (KBr,  = cm−1): 2120
0.01
(N3), 1684 (C=O).
3.89
7.27
6.94
7.92
7.87
6.99
002001.FID
2.01
1.91
7.5
Chemical Shift (ppm)
7.0
2.01
8.5
8.0
6.99
6.94
7.27
7.92
7.87
4.51
8.0
1.91
7.5
7.0
2.08
6.5
6.0
5.5
5.0
2.86
4.5
4.0
Chemical Shift (ppm)
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0
-0.5
Figure S3. 1H NMR spectrum (200 MHz, CDCl3) of 2-azido-1-(4-methoxyphenyl)ethanone
(6)
Wavenumber (cm-1)
Figure S4. IR spectrum (KBr) of 2-azido-1-(4-methoxyphenyl)ethanone (6)
2-azido-1-(4-bromophenyl)ethanone (7): 1H NMR (400 MHz, CDCl3): δ 4.53 (s, 2H); 7.66
7.27
(d, 2H, J 8.0 Hz); 7.79 (d, 2H, J 8.0 Hz). IR (KBr,  = cm−1): 2102 (N3), 1690 (C=O).
7.65
1.99
7.5
7.65
7.80
7.78
7.67
7.8
7.7
7.6
Chemical Shift (ppm)
4.53
7.78
7.80
2.02
7.9
7.67
9D929_BROMOCITOAZIDA.001.esp
2.02 1.99
9.0
8.5
8.0
7.5
2.00
7.0
6.5
6.0
5.5
5.0
4.5
4.0
Chemical Shift (ppm)
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0
-0.5
Figure S5. 1H NMR spectrum (400 MHz, CDCl3) of 2-azido-1-(4-bromophenyl)ethanone
(7)
Wavenumber (cm-1)
Figure S6. IR spectrum (KBr) of 2-azido-1-(4-bromophenyl)ethanone (7)
2-azido-1-(4-nitrophenyl)ethanone (8): 1H NMR (400 MHz, CDCl3): δ 4.64 (s, 2H); 8.08 (d,
2H, J 8.0 Hz); 8.31 (d, 2H, J 8.0 Hz). IR (KBr,  = cm−1): 2105 (N3), 1706 (C=O), 1554,
1349.
Figure S7. 1H NMR spectrum (400 MHz, CDCl3) of 2-azido-1-(4-nitrophenyl)ethanone (8)
Wavenumber (cm-1)
Figure S8. IR spectrum (KBr) of 2-azido-1-(4-nitrophenyl)ethanone (8)
Preparation of ()-β-azidophenylethanols 9-12
The ketones 5-8 (5.0 mmol), NaBH4 (5.5 mmol) and methanol (10 mL) were added to a 25
mL flask equipped with a magnetic stirrer. The mixtures were stirred for 1 h at 0 oC. After
that, the reaction mixtures were quenched by the addition of water (5 mL), the methanol was
removed under vacuum and the residue was extracted with EtOAc (3 x 20 mL). The
combined organic phases were dried over MgSO4 and then filtered. The organic solvent was
evaporated under reduced pressure and the residue was purified by silica gel column
chromatography using hexane and ethyl acetate as eluent to produce racemic βazidophenylethanols 9-12.
2-azido-1-phenylethanol (9): 1H NMR (200 MHz, CDCl3): δ (ppm) 2.49 (s, 1H); 3.35-3.52
(m, 2H); 4.84 (dd, 1H, J 8.0, 4.0 Hz); 7.35 (m, 5H). 13C RMN (50 MHz, CDCl3): 58.0, 73.3,
7.35
125.9, 128.6, 140.5. IR (KBr,  = cm−1): 3400 (OH), 2103 (N3).
001001.FID
3.35
3.38
1.00
2.29
4.85
4.80
Chemical Shift (ppm)
3.50
3.45
3.40
Chemical Shift (ppm)
3.35
5.02
8.5
8.0
1.00
7.5
7.0
6.5
6.0
5.5
5.0
0.00
3.38
3.35
3.52
3.48
2.49
4.87
4.85
4.84
4.81
3.46
3.44
3.42
3.42
4.90
3.42
3.42
3.44
3.46
3.52
3.48
4.81
4.87
4.85
001001.FID
4.84
001001.FID
2.29
4.5
4.0
Chemical Shift (ppm)
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0
-0.5
128.64
125.87
Figure S9. 1H NMR spectrum (200 MHz, CDCl3) of 2-azido-1-phenylethanol (9)
125.87
128.29
001001.FID
128.29
128.64
001001.FID
126
125
57.98
128
127
Chemical Shift (ppm)
140.51
77.00
73.34
129
192
184
176
168
160
152
144
136
128
120
112
104
96
88
Chemical Shift (ppm)
80
72
64
56
48
40
32
24
16
8
Figure S10. 13C NMR spectrum (50 MHz, CDCl3) of 2-azido-1-phenylethanol (9)
0
-8
Wavenumber (cm-1)
Figure S11. IR spectrum (KBr) of 2-azido-1-phenylethanol (9)
2-azido-1-(4-methoxyphenyl)ethanol (10): 1H NMR (200 MHz, CDCl3): δ (ppm) 2.19 (s,
1H); 3.36-3.54 (m, 3H); 3.82 (s, 3H); 4.84 (dd, 1H, J 8.0, 4.0 Hz); 6.90 (d, 2H, J 8.0 Hz);
7.30 (d, 2H, J 8.0 Hz).
13
C RMN (50 MHz, CDCl3): 55.0, 57.6, 72.6, 113.8, 127.0, 132.7,
3.82
159.2. IR (KBr,  = cm−1): 3418 (OH), 2103 (N3).
ALCOOL METOXI AZIDA 2.ESP
3.38
3.36
3.51
3.54
3.48
3.42
3.45
ALCOOL METOXI AZIDA 2.ESP
2.90
7.5
3.55
3.50
3.45
3.40
3.35
Chemical Shift (ppm)
3.30
3.25
3.20
3.15
1.15
6.5
6.0
5.5
5.0
3.24
3.38
3.36
2.19
3.48
3.45
3.42
2.35
7.0
3.54
4.87
4.85
4.83
4.81
2.26
8.0
3.60
6.94
6.89
7.33
7.28
3.65
2.90
4.5
4.0
3.5
Chemical Shift (ppm)
3.0
2.5
2.0
1.5
1.0
0.5
0
Figure S12. 1H NMR spectrum (200 MHz, CDCl3) of 2-azido-1-(4-methoxyphenyl)ethanol
(10)
113.78
159.22
132.72
55.02
72.61
57.59
127.02
9D930_AZIDO ALCOOL.002.esp
192
184
Figure S13.
13
176
168
160
152
144
136
128
120
112
104
96
88
Chemical Shift (ppm)
80
72
64
56
48
40
32
24
16
8
0
C NMR spectrum (50 MHz, CDCl3) of 2-azido-1-(4-methoxyphenyl)ethanol
(10)
Wavenumber (cm-1)
Figure S14. IR spectrum (KBr) of 2-azido-1-(4-methoxyphenyl)ethanol (10)
2-azido-1-(4-bromophenyl)ethanol (11): 1H NMR (200 MHz, CDCl3): δ (ppm) 3.45 (d, 2H,
J 6,0 Hz); 4.85 (dd, 1H, J 12.0, 6.0 Hz); 7.26 (d, 2H, J 8.0 Hz); 7.50 (d, 2H, J 8.0 Hz). 13C
RMN (50 MHz, CDCl3): 57.9, 72.7, 122.2, 127.6, 131.8, 139.4, 159.4. IR (KBr,  = cm−1)
7.28
3406 (OH), 2103 (N3).
4.88
4.82
4.85
7.24 7.27
7.53
3.46
3.43
7.49
001001.FID
2.13 2.34
9.0
8.5
8.0
7.5
1.00
7.0
6.5
6.0
5.5
2.24
5.0
4.5
4.0
Chemical Shift (ppm)
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0
Figure S15. 1H NMR spectrum (200 MHz, CDCl3) of 2-azido-1-(4-bromophenyl)ethanol
139.45
122.23
72.75
57.91
131.79
001001.FID
127.61
(11)
208
200
192
184
176
168
160
152
144
136
128
120
112
104
96
Chemical Shift (ppm)
88
80
72
64
56
48
40
32
24
16
8
0
Figure S16. 13C NMR spectrum (50 MHz, CDCl3) 2-azido-1-(4-bromophenyl)ethanol (11)
Wavenumber (cm-1)
Figure
S17.
IR
spectrum
(KBr)
of
2-azido-1-(4-bromophenyl)ethanol
(11)
2-azido-1-(4-nitrophenyl)ethanol (12): 1H NMR (400 MHz, CDCl3): δ (ppm) 1.95 (s, 1H);
3.39-3.47 (m, 2H); 4.97 (dd, 1H, J 8.0, 4.0 Hz); 7.51 (d, 2H, J 8.0 Hz); 8.08 (d, 2H, J 8.0
Hz).
C RMN (100 MHz, CDCl3): 57.1, 72.0, 123.3, 126.6, 147.0, 148.0 IR (KBr,  =
13
7.52
cm−1): 3401 (OH), 2108 (N3).
9D930_NITRO AZIDO ALCOOL.001.esp
3.44
4.975
4.950
Chemical Shift (ppm)
3.39
3.39
3.41
3.40
3.46
3.47
3.42 3.43
1.00
5.000
1.95
3.44
8.09
8.07
3.43
3.42
3.44
3.44
4.96
4.95
4.98
4.97
9D930_NITRO AZIDO ALCOOL.001.esp
7.50
9D930_NITRO AZIDO ALCOOL.001.esp
2.08
3.39
3.39
3.450
3.425
3.400
Chemical Shift (ppm)
7.27
3.47 3.46
4.98
4.97
4.96
4.95
3.475
1.83
9.5
9.0
8.5
8.0
2.00
1.00
7.5
7.0
6.5
6.0
5.5
5.0
4.5
Chemical Shift (ppm)
2.08
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0
-0.5
126.60
Figure S18. 1H NMR spectrum (400 MHz, CDCl3) of 2-azido-1-(4-nitrophenyl)ethanol (15)
148.06
147.08
57.14
72.01
123.33
9D930_NITRO AZIDO ALCOOL.002.esp
192
184
176
168
160
152
144
136
128
120
112
104
96
88
Chemical Shift (ppm)
80
72
64
56
48
40
32
24
16
8
0
-8
Figure S19. 13C NMR spectrum (100 MHz, CDCl3) 2-azido-1-(4-nitrophenyl)ethanol (12)
Wavenumber (cm-1)
Figure S20. IR spectrum (KBr) of 2-azido-1-(4-nitrophenyl)ethanol (12)
Chromatograms of the reduction of α-azido ketones 5-8 using whole cells of fungi.
Bioreduction (±)-2-azido-1-phenyltanone (5) by whole cells of fungi
M. racemosus CBMAI 847
c = 10%
t = 12 min
A. sclerotiorum CBMAI 849
A
c = 90%
72% ee
t = 26 min
27 min
c = 2%
t = 12 min
C
B. felina CBMAI 738
B
c = 98%
85% ee
t = 26 min
27 min
P. raistrickii CBMAI 931
c = 100%
80% ee
c = 10%
t = 12 min
t = 26 min
27 min
D
c = 90%
90% ee
t = 26 min
27 min
Figure S21. Chiral analyses obtained by HPLC chromatograms. Conditions for HPLC
analysis in 10% isopropanol, 90% hexane, flow 0.5 mL/min, 190 nm. (A) M. racemosus
CBMAI 847 (B) A. sclerotiorum CBMAI 849 (C) B. felina CBMAI 738 (D) P. raistrickii
CBMAI 931
Bioreduction (±)-2-azido-1-(4-methoxyphenyl)etanone (6) by whole cells of fungi
A
t = 33 min
C. cladosporiodes CBMAI 857
B
A. sclerotiorum CBMAI 849
c = 100%
>99% ee
c = 100%
>99% ee
t = 33 min
C
P. raistrickii CBMAI 931
c = 32%
t = 23 min
t = 33 min
c = 68%
>99% ee
P. citrinum CBMAI 1186
t = 33 min
D
c = 99%
>99% ee
Figure S22. Chiral analyses obtained by HPLC chromatograms. Conditions for HPLC
analysis in 10% isopropanol, 90% hexane, flow 0.5 mL/min, 190 nm. (A) C.
cladosporioides CBMAI 857 (B) A. sclerotiorum CBMAI 931 (C) P. raistrickii CBMAI
849 (D) P. citrinum CBMAI 1186
Bioreduction (±)-2-azido-1-(4-bromophenyl)etanone (7) by whole cells of fungi
M. racemosus CBMAI 847
c = 21%
t = 24 min
28 min
A
c = 79%
29% ee
t = 15 min
C. cladosporiodes CBMAI 857
A. sclerotiorum CBMAI 849
c = 25%
C
t = 24 min
28 min
t = 24 min
28 min
B
t = 15 min
c = 75%
39% ee
D
P. raistrickii CBMAI 931
t = 15 min
c = 98%
44% ee
c = 2%
c = 68%
t = 15 min
c = 32%
70% ee
t = 24 min
28 min
Figure S23. Chiral analyses obtained by HPLC chromatograms. Conditions for HPLC
analysis in 10% isopropanol, 90% hexane, flow 0.5 mL/min, 190 nm. (A) M. racemosus
CBMAI 847 (B) A. sclerotiorum CBMAI 849 (C) C. cladosporioides CBMAI 857 (D) P.
raistrickii CBMAI 931
Bioreduction (±)-2-azido-1-(4-nitrophenyl)etanone (8) by whole cells of fungi
t = 21 min
23 min
P. citrinum CBMAI 1186
A
t = 21 min
23 min
P. oxalicum CBMAI 1185
c = 95%
28% ee
B
c = 96%
51% ee
c = 5%
c = 4%
C
P. chrysogenum CBMAI 1199
t = 21 min
23 min
c = 98%
41% ee
Figure S24. Chiral analyses obtained by HPLC chromatograms. Conditions for HPLC
analysis in 10% isopropanol, 90% hexane, flow 0.5 mL/min, 254 nm. (A) P. citrinum
CBMAI 1186 (B) P. oxalicum CBMAI 1185 (C) P. chrysogenum 1199