Part E | 28.2
692 Part E Marine Microbiology and Biotechnology
Table 28.1 Growth of marine fungi on a solid medium with various amounts of phenylacetonitrile 1 (32
ı C, 8 days)
Marine fungi
Colony diameter (cm)
5 L
(0:04 mmol)
10 L
(0:08 mmol)
15 L
(0:12 mmol)
20 L
(0:17 mmol)
Penicillium miczynskii CBMAI 930
a
–
–
–
Penicillium raistrickii CBMAI 931
1.5
1.5
1.5
0.5
Aspergillus sydowii CBMAI 933
1.5
2.0
2.0
a
Aspergillus sydowii CBMAI 934
1.5
1.8
1.8
a
Aspergillus sydowii CBMAI 935
1.5
2.0
2.0
0.5
Bionectria sp. CBMAI 936
2.5
3.5
3.5
a
Penicillium decaturense CBMAI 1234
1.5
1.5
1.5
a
Penicillium raistrickii CBMAI 1235
1.0
1.0
1.0
a
Cladosporium sp. CBMAI 1237
a
–
–
–
Aspergillus sydowii CBMAI 1241
1.2
1.5
1.5
a
Penicillium oxalicum CBMAI 1185
a
–
–
–
Penicillium citrinum CBMAI 1186
a
–
–
–
a No colony formed
Table 28.2 Growth of marine fungi in liquid mineral medium (100:0 mL) with various amounts of phenylacetonitrile 1
(32
ı C, 124 rpm, 8 days)
Volume of phenylacetonitrile 1 (L)/Erlenmeyer flask
Marine fungi
20:0 (0:17 mmol)
40:0 (0:34 mmol)
60:0 (0:52 mmol)
Total cell dry weight (g)
Aspergillus sydowii CBMAI 935
0.61
0.75
0.90
Aspergillus sydowii CBMAI 934
0.61
0.66
0.94
Aspergillus sydowii CBMAI 1241
0.72
0.95
1.10
Aspergillus sydowii CBMAI 933
0.69
0.72
0.72
Bionectria sp. CBMAI 936
0.46
0.59
0.63
Penicillium decaturense CBMAI 1234
0.41
1.10
1.46
Penicillium raistrickii CBMAI 931
1.10
1.10
1.10
Penicillium raistrickii CBMAI 1235
0.88
0.95
1.28
under vacuum. The extracts obtained were analyzed
by GC-FID and GC-MS. The extracts obtained from
the filtered medium and the mycelia were combined
and purified by flash column chromatography (CC)
over silica gel to yield 2-hydroxyphenylacetic acid 1a
(51% yield). The spectroscopic data of the isolated 2hydroxyphenylacetic acid 1a were in agreement with
those reported in the literature [28.55].
28.2.5 Biotransformation and Isolation
of Phenylacetonitrile Derivatives 2–4
by A. sydowii CBMAI 934
Small slices of solid medium (0:5 0:5 cm) bearing mycelia of the fungal strain were cut from
the solid stock cultures and used to inoculate
100:0 mL of liquid culture mineral medium with glucose (15:0 g L
1 ), containing 60 L of phenylacetonitrile 1 (0:52 mmol), in 250 mL Erlenmeyer flasks.
The marine fungus was grown at 32
ı C on a rotary shaker (124 rpm). After growth of the strain
in liquid medium for 96 h, the following nitriles
were added separately to each experiment: 4-fluorophenylacetonitrile 2 (20:0 L (0:17 mmol), 40 L
(0:34 mmol), 60 L (0:51 mmol)), 4-chlorophenylacetonitrile 3 (20:0 L (0:16 mmol), 40:0 L (0:32 mmol),
60:0 L (0:48 mmol)), and 4-methoxyphenylacetonitrile 4 (20:0 L (0:15 mmol), 40:0 L (0:30 mmol),
60:0 L (0:45 mmol)). The biotransformations of nitriles 2–4 were monitored every 24 h by collecting
692 Part E Marine Microbiology and Biotechnology
Table 28.1 Growth of marine fungi on a solid medium with various amounts of phenylacetonitrile 1 (32
ı C, 8 days)
Marine fungi
Colony diameter (cm)
5 L
(0:04 mmol)
10 L
(0:08 mmol)
15 L
(0:12 mmol)
20 L
(0:17 mmol)
Penicillium miczynskii CBMAI 930
a
–
–
–
Penicillium raistrickii CBMAI 931
1.5
1.5
1.5
0.5
Aspergillus sydowii CBMAI 933
1.5
2.0
2.0
a
Aspergillus sydowii CBMAI 934
1.5
1.8
1.8
a
Aspergillus sydowii CBMAI 935
1.5
2.0
2.0
0.5
Bionectria sp. CBMAI 936
2.5
3.5
3.5
a
Penicillium decaturense CBMAI 1234
1.5
1.5
1.5
a
Penicillium raistrickii CBMAI 1235
1.0
1.0
1.0
a
Cladosporium sp. CBMAI 1237
a
–
–
–
Aspergillus sydowii CBMAI 1241
1.2
1.5
1.5
a
Penicillium oxalicum CBMAI 1185
a
–
–
–
Penicillium citrinum CBMAI 1186
a
–
–
–
a No colony formed
Table 28.2 Growth of marine fungi in liquid mineral medium (100:0 mL) with various amounts of phenylacetonitrile 1
(32
ı C, 124 rpm, 8 days)
Volume of phenylacetonitrile 1 (L)/Erlenmeyer flask
Marine fungi
20:0 (0:17 mmol)
40:0 (0:34 mmol)
60:0 (0:52 mmol)
Total cell dry weight (g)
Aspergillus sydowii CBMAI 935
0.61
0.75
0.90
Aspergillus sydowii CBMAI 934
0.61
0.66
0.94
Aspergillus sydowii CBMAI 1241
0.72
0.95
1.10
Aspergillus sydowii CBMAI 933
0.69
0.72
0.72
Bionectria sp. CBMAI 936
0.46
0.59
0.63
Penicillium decaturense CBMAI 1234
0.41
1.10
1.46
Penicillium raistrickii CBMAI 931
1.10
1.10
1.10
Penicillium raistrickii CBMAI 1235
0.88
0.95
1.28
under vacuum. The extracts obtained were analyzed
by GC-FID and GC-MS. The extracts obtained from
the filtered medium and the mycelia were combined
and purified by flash column chromatography (CC)
over silica gel to yield 2-hydroxyphenylacetic acid 1a
(51% yield). The spectroscopic data of the isolated 2hydroxyphenylacetic acid 1a were in agreement with
those reported in the literature [28.55].
28.2.5 Biotransformation and Isolation
of Phenylacetonitrile Derivatives 2–4
by A. sydowii CBMAI 934
Small slices of solid medium (0:5 0:5 cm) bearing mycelia of the fungal strain were cut from
the solid stock cultures and used to inoculate
100:0 mL of liquid culture mineral medium with glucose (15:0 g L
1 ), containing 60 L of phenylacetonitrile 1 (0:52 mmol), in 250 mL Erlenmeyer flasks.
The marine fungus was grown at 32
ı C on a rotary shaker (124 rpm). After growth of the strain
in liquid medium for 96 h, the following nitriles
were added separately to each experiment: 4-fluorophenylacetonitrile 2 (20:0 L (0:17 mmol), 40 L
(0:34 mmol), 60 L (0:51 mmol)), 4-chlorophenylacetonitrile 3 (20:0 L (0:16 mmol), 40:0 L (0:32 mmol),
60:0 L (0:48 mmol)), and 4-methoxyphenylacetonitrile 4 (20:0 L (0:15 mmol), 40:0 L (0:30 mmol),
60:0 L (0:45 mmol)). The biotransformations of nitriles 2–4 were monitored every 24 h by collecting