Biotransformation of Nitriles by Marine Fungi 28.2 Experimental Methods 695
Part E | 28.3
Preparation of liquid culture media
100 mL of mineral medium + phenylacetonitrile 1
1. Inoculum of microorganism
2. Incubation for 96 h at 32 °C and 124 rpm
1. Experiments pooled
2. Addition of cyclohexenylacetonitrile 5 (60 µL)
3. Incubation for 96 h at 32 °C and 124 rpm
1. Filtration
2. Exctration with AcOEt
3. Purification by CC-flash
4. GC-MS,
1 H and
13 C NMR
Erlenmeyer flask of 250 mL
Erlenmeyer flask of 1 L
Product formation
Cyclohexenylacetic acid
(m = 74 mg, 28%)
Reaction followed by GC-FID
Culture of microorganism
(96 h of microbial growth)
Fig. 28.33 Biotransformation of cyclohexenylacetonitrile 5 by A. sydowii CBMAI 934
28.2.9 Biotransformation
of Benzonitrile 6
and 2-Cyanopyridine 14
Two different experiments were carried out with these
nitriles. In the first, after the fungus had grown in
100:0 mL of the liquid culture mineral medium in Erlenmeyer flasks (250 mL) for 96 h in the presence of
glucose (15:0 g L
1 ) and phenylacetonitrile 1 (60:0 L,
0:52 mmol), benzonitrile 6 (20:0 L, 0:19 mmol) and
2-cyanopyridine 14 (20:0 L, 0:21 mmol) were added
simultaneously. In the second experiment, after the fungal growth in the presence of glucose (15:0 g L
1 )
and phenylacetonitrile 1 (60:0 L, 0:52 mmol) for 96 h,
2-cyanopyridine 14 (20:0 L, 0:21 mmol) alone was
added to the liquid culture mineral medium. The reactions were monitored every 24 h by collecting 2:0 mL
samples, which were extracted by stirring with EtOAc
(2:0 mL), centrifuged (6000 rpm, 6 min), and analyzed
by GC-FID and GC-MS.
Part E | 28.3
Preparation of liquid culture media
100 mL of mineral medium + phenylacetonitrile 1
1. Inoculum of microorganism
2. Incubation for 96 h at 32 °C and 124 rpm
1. Experiments pooled
2. Addition of cyclohexenylacetonitrile 5 (60 µL)
3. Incubation for 96 h at 32 °C and 124 rpm
1. Filtration
2. Exctration with AcOEt
3. Purification by CC-flash
4. GC-MS,
1 H and
13 C NMR
Erlenmeyer flask of 250 mL
Erlenmeyer flask of 1 L
Product formation
Cyclohexenylacetic acid
(m = 74 mg, 28%)
Reaction followed by GC-FID
Culture of microorganism
(96 h of microbial growth)
Fig. 28.33 Biotransformation of cyclohexenylacetonitrile 5 by A. sydowii CBMAI 934
28.2.9 Biotransformation
of Benzonitrile 6
and 2-Cyanopyridine 14
Two different experiments were carried out with these
nitriles. In the first, after the fungus had grown in
100:0 mL of the liquid culture mineral medium in Erlenmeyer flasks (250 mL) for 96 h in the presence of
glucose (15:0 g L
1 ) and phenylacetonitrile 1 (60:0 L,
0:52 mmol), benzonitrile 6 (20:0 L, 0:19 mmol) and
2-cyanopyridine 14 (20:0 L, 0:21 mmol) were added
simultaneously. In the second experiment, after the fungal growth in the presence of glucose (15:0 g L
1 )
and phenylacetonitrile 1 (60:0 L, 0:52 mmol) for 96 h,
2-cyanopyridine 14 (20:0 L, 0:21 mmol) alone was
added to the liquid culture mineral medium. The reactions were monitored every 24 h by collecting 2:0 mL
samples, which were extracted by stirring with EtOAc
(2:0 mL), centrifuged (6000 rpm, 6 min), and analyzed
by GC-FID and GC-MS.