Biotransformation of Nitriles by Marine Fungi 28.2 Experimental Methods 693
Part E | 28.2
Table 28.3 Biotransformation of phenylacetonitrile 1 and its derivatives 2–4 by marine fungus A. sydowii CBMAI 934
a
CN
OH
OH
O
R 1
OH
O
R 1
R 3
R 2
CN
R 3
R 2
A. sydowii
CBMAI 934
32 °C, 124 rpm
1
+
+
1a
2a-4a
R 1 = R 2 = H, R 3 = F (2)
R 1 = R 2 = H, R 3 = Cl (3)
R 1 = R 2 = H, R 3 = OMe (4)
Time (h)
c (%)
c (%)
c (%)
c (%) [Yield] (%)
#
1
2
1a
2a
24
100
–
–
–
96 b
–
100
100
–
120
–
–
12
88 [28.51]
#
1
3
1a
3a
24
100
–
–
–
96 b
–
100
100
–
120
–
–
10
90 [28.55]
#
1
4
1a
4a
24
100
–
–
–
96 b
–
100
100
–
120
–
50
28
22
192
–
–
12
88 [28.43]
a (Sect. 28.2).
b The nitriles 2–4 were added (Sect. 28.2).
c D concentration of unreacted nitriles 1–4 determined by GC-FID analysis.
c D conversion determined by GC-FID analysis.
2.0 mL of samples, which were extracted by stirring
with EtOAc (Ethyl Acetate) (2:0 mL) and then centrifuged (6; 000 rpm, 6 min) and analyzed by GC-FID
(Fig. 28.32).
28.2.6 Biotransformation
of Cyclohexenylacetonitrile 5
by A. sydowii CBMAI 934
Initially, the fungal strain was cultured as described in
Sect. 28.2.5. After the initial growth of the mycelia,
cyclohexenylacetonitrile 5 (20:0 L (0:16 mmol),
40:0 L (0:32 mmol), and 60:0 L (0:48 mmol)) was
added to the flasks. In a second experiment, after fungal
growth as in Sect. 28.2.5, the mycelia were pooled
(3 in 1) and transferred to a 1 L Erlenmeyer flask,
and then further cyclohexenylacetonitrile 5 (60:0 L,
0:48 mmol) was added. The biotransformation of nitrile
5 was monitored every 24 h for 96 h by collecting
2:0 mL samples, which were extracted by stirring with
ethyl acetate (2:0 mL), centrifuged (6000 rpm, 6 min)
and analyzed by GC-FID (Fig. 28.33).
28.2.7 Biotransformation
of Aromatic Nitriles
After the growth of the fungal strain in 100:0 mL
liquid culture mineral medium supplemented with glucose (15:0 g=L) and phenylacetonitrile 1 (60:0 L, 0:52
mmol) for 96 h in 250 mL Erlenmeyer flasks, the respective aromatic nitriles were added: benzonitrile 6
(20:0 L, 0:19 mmol), 2-chlorobenzonitrile 7 (20:0mg,
0:15 mmol), 4-hydroxybenzonitrile 8 (20:0 mg, 0:17
mmol), 4-methoxybenzonitrile 9 (20:0mg, 0:20 mmol),
3,5-dimethoxybenzonitrile 10 (20:0mg, 0:12 mmol), 2nitrobenzonitrile 11 (20:0mg, 0:14 mmol), 3-nitrobenzonitrile 12 (20:0mg, 0:13 mmol) and 4-nitrobenzonitrile 13 (20:0mg, 0:14 mmol). The reactions were monitored every 24 h by collecting 2:0 mL aliquots, which
were extracted by stirring with EtOAc (2:0 mL), fol-
Part E | 28.2
Table 28.3 Biotransformation of phenylacetonitrile 1 and its derivatives 2–4 by marine fungus A. sydowii CBMAI 934
a
CN
OH
OH
O
R 1
OH
O
R 1
R 3
R 2
CN
R 3
R 2
A. sydowii
CBMAI 934
32 °C, 124 rpm
1
+
+
1a
2a-4a
R 1 = R 2 = H, R 3 = F (2)
R 1 = R 2 = H, R 3 = Cl (3)
R 1 = R 2 = H, R 3 = OMe (4)
Time (h)
c (%)
c (%)
c (%)
c (%) [Yield] (%)
#
1
2
1a
2a
24
100
–
–
–
96 b
–
100
100
–
120
–
–
12
88 [28.51]
#
1
3
1a
3a
24
100
–
–
–
96 b
–
100
100
–
120
–
–
10
90 [28.55]
#
1
4
1a
4a
24
100
–
–
–
96 b
–
100
100
–
120
–
50
28
22
192
–
–
12
88 [28.43]
a (Sect. 28.2).
b The nitriles 2–4 were added (Sect. 28.2).
c D concentration of unreacted nitriles 1–4 determined by GC-FID analysis.
c D conversion determined by GC-FID analysis.
2.0 mL of samples, which were extracted by stirring
with EtOAc (Ethyl Acetate) (2:0 mL) and then centrifuged (6; 000 rpm, 6 min) and analyzed by GC-FID
(Fig. 28.32).
28.2.6 Biotransformation
of Cyclohexenylacetonitrile 5
by A. sydowii CBMAI 934
Initially, the fungal strain was cultured as described in
Sect. 28.2.5. After the initial growth of the mycelia,
cyclohexenylacetonitrile 5 (20:0 L (0:16 mmol),
40:0 L (0:32 mmol), and 60:0 L (0:48 mmol)) was
added to the flasks. In a second experiment, after fungal
growth as in Sect. 28.2.5, the mycelia were pooled
(3 in 1) and transferred to a 1 L Erlenmeyer flask,
and then further cyclohexenylacetonitrile 5 (60:0 L,
0:48 mmol) was added. The biotransformation of nitrile
5 was monitored every 24 h for 96 h by collecting
2:0 mL samples, which were extracted by stirring with
ethyl acetate (2:0 mL), centrifuged (6000 rpm, 6 min)
and analyzed by GC-FID (Fig. 28.33).
28.2.7 Biotransformation
of Aromatic Nitriles
After the growth of the fungal strain in 100:0 mL
liquid culture mineral medium supplemented with glucose (15:0 g=L) and phenylacetonitrile 1 (60:0 L, 0:52
mmol) for 96 h in 250 mL Erlenmeyer flasks, the respective aromatic nitriles were added: benzonitrile 6
(20:0 L, 0:19 mmol), 2-chlorobenzonitrile 7 (20:0mg,
0:15 mmol), 4-hydroxybenzonitrile 8 (20:0 mg, 0:17
mmol), 4-methoxybenzonitrile 9 (20:0mg, 0:20 mmol),
3,5-dimethoxybenzonitrile 10 (20:0mg, 0:12 mmol), 2nitrobenzonitrile 11 (20:0mg, 0:14 mmol), 3-nitrobenzonitrile 12 (20:0mg, 0:13 mmol) and 4-nitrobenzonitrile 13 (20:0mg, 0:14 mmol). The reactions were monitored every 24 h by collecting 2:0 mL aliquots, which
were extracted by stirring with EtOAc (2:0 mL), fol-