J. CHEM. RESEARCH (S), 1999 397
Table 3 1 NMR data of compounds 1^7 (400 MHz, CDCl3, 1
and 3, 300 MHZ, 4, in CD3OD)a
6 was particularly intriguing arising from concomitant
oxidation and reduction in the same reaction vessel. This
presumably results from compartmentalisation of the
enzymes responsible within the cell or a switch between
H
1
2
3
4
5
6
7
1
2
1.90m
1.50mb
1.70mb
1.30m
1.70mb
2.00m
1.80m
3.09m
1.27m
1.60m
1.43m
2.36m
1.98m
2.25m
1.90m
1.50mb
1.70mb
1.30mb
1.70mb
2.00m
1.80m
1.90m
1.30mb
1.60mb
1.30mb
1.60mb
2.00m
1.70m
3.20m
1.30m
1.60m
1.50m
2.40m
2.01m
2.40m
2.00m
1.30mb
1.50mb
1.70mb
2,00mb
1.90m
1.70m
ö
oxidative
and
reductive
metabolism
during
the
1.35mb
2.50mb
1.35mb
2.50mb
1.90m
1.75dd
(10.5, 6.5)
0.21dd
fermentation. Compound 7 has been obtained by micro-
biological methods for the ¢rst time. Low conversions
are normally attained for these biotransformations; in most
reactions reported here, 40% of the starting materials were
recovered, so that, yields of products based on starting
materials consumed were generally in the range 20^57%.
3
4
5
6
7
0.12dd
(9.0)
0.66
0.36dd
(9.1, 11.1) (9.5)
0.64ddd
(4.7, 9.1, (6.0, 9.5,
0.32dd
0.28dd
(9.1)
0.77m
0.55dd
0.41dd
(9.5, 11.0) (9.2, 11.2) (9.3, 10.6)
0.80ddd 0.72ddd 0.72ddd
(4.9, 9.5, (5.2, 9.2, (5.2, 9.3,
0.83ddd
13.7)
10.5)
11.8)
12.0)
12.0)
8
9
1.50mb
1.70mb
1.50mb
1.70mb
1.63m
1.83m
2.44m
2.53m
1.50mb
1.70mb
1.50mb
1.70mb
1.45mb
1.65mb
1.45mb
1.65mb
1.65m
1.80m
2.50m
2.70m
1.50mb
1.70mb
1.40mb
1.60mb
1.35mb
1.90m
3.01ddd
(3.7, 5.1,
13.6)
Experimental
1H and 13C NMR spectra were determined at 400 or 300 and 100 or
75 MHz respectively. Chemical shifts were measured in ppm
down¢eld of TMS and coupling constants (J) in Hz (see Tables 2-4).
IR spectra were determined in CH2CI2. Mass spectra were recorded
on a TRIO1000 instrument. TLC was performed on Merck 60
F254 precoated silica plates and spots were detected by spraying with
a solution of 1.5% NH4Mo2O2, 1% CeIVSO4 and 10% H2SO4
followed by charring. Flash chromatography was performed with
Si-60 (Merck, 40^63 lm). The microorganisms were precultivated
at 28 8C and 120 rpm in 100 mL conical ¯asks containing 100 mL
of the following medium: 5% malt extract and distilled water. After
96 h, 100 mg of substrate dissolved in 0.5 mL EtOH were added to
the cultures. Every day, starting 24 h after the substrate addition,
samples were taken and analysed. Culture medium and mycelia were
both separated by ®ltration and extracted (Â3) with EtOAc. The
solvent was evaporated and the crude extract separated on Si-60
columns with the appropriate light petroleum±EtOAc gradient.
10b,14-Dihydroxy-allo-aromadendrane 1. [a]2D0 4:53 (CHCl3; c 1).
2,48mb
ö
10
12 1.01s
ö
ö
ö
ö
ö
3.90
1.04s
1.15s
3.50d
(12.0)
3.60d
(12.0)
1.10s
1.20s
1.09s
0.99s
13 1.03s
1.00s
3.26d
(11.0)
3.47d
(11.0)
3.20d
(11.0)
3.40d
(11.0)
3.24d
(10.8)
3.42d
(10.8)
0.96s
14 3.41d
(10.9)
0.94d(6.8) 3.29d
(10.5)
3.20d
(11.0)
3.30d
(11.0)
0.98d(6.9) 0.96d(6.8) 0.94d(6.6)
3.28d
(10.9)
3.42d
910.5)
0.95d(7.0) 0.96d(5.5)
15 0.93d(6.6)
OH
ö
ö
ö
ö
2.95s br
aThe data were consistent with those reported in the literature,3
however, in this work a detailed assignment was accomplished.
bThe assignments for these signals within the same column may
be interchanged.
8
1
nmax(CH2Cl2 cast)/cm 3292, 2950, 2867, 1453, 1029. EIMS m/z
(rel. int.): [M H2O 220.15 (15), 207.15 (21), 203.15 (40), 189.05
(60), 81.95 (14), 40.90 (100).
Allo-Aromadendrone 2. [a]2D0 8:32 (CHCl3; c 0.8). n(CH2Cl2
1
cast)/cm 2961, 2874, 1733, 1695, 1459, 1000. EIMS m/z (rel. int.):
Table 4 NOE observed for compounds 2 and 6
[M] 206 (15), 163 (12), 83 (38), 69 (100).
2
6
10b,13,14-Trihydroxy-allo-aromadendrane
3.
nmaxꢀCH2Cl2
1
cast)/cm
3335, 2957, 2871, 1460, 1020. CIMS m/z. Found:
H
6
10
Enhanced protons
7, 14, 13
6, 14, 13
Enhanced protons
7, 10, 13
6, 7
[M H2O NH4 , 254.21210. C15H28NO2 requires 254.21200.
10b,12,14-Trihydroxy-allo-aromadendrane
4.
nmaxꢀCH2Cl2
1
cast)/cm
3399, 2958, 2874, 1457, 1020. CIMS m/z. Found:
M NH4 , 272.22262. C15H30NO3 requires 272.22257.
13-Hydroxy-allo-aromadendrone 5. nmax (CH2Cl2 cast)/cm 1 3615,
We are grateful to CAPES-Brazil for ¢nancial support
2959, 1696, 1458, 1015. EIMS m/z. Found: M , 222.16180.
C
14H22O2 requires 222.16199.
to Denis P. de Lima. We are also in debt to Dr Joao
M. de Siqueira (for collecting D. glabriuscula and providing
information about bioactivity of the substrates) and, Dr
Mike Paylor, for growing Rhizopus sp. We also thank
Lucineia Vizzotto for samples of the substrates over the
duration of this work, and Dr Paul A. Leonard for help
on the use of various NMR techniques.
10b,13-Dihydroxy-allo-aromadendrane 6. nmax(CH2Cl2 cast)/
1
cm 3615, 3460, 3007, 2957, 2874, 1466, 1011. CIMS m/z. Found:
M NH4 H2O , 224.20181. C14H26NO requires 224.20144.
1
1-Hydroxy-allo-aromadendrone 7. nmax(CH2Cl2 cast)/cm 3597,
3439, 2970, 2858, 1703, 1676, 1462, 1261, 1096, 1027. EIMS m/z.
Found: M , 222.16202. C14H22O2 requires 222.16199.
Table 2 13C NMR data of compounds 1^7 (100 MHz, CDCl3, 1
375 MHz, 4, in CD3OD)a
Received, 15th February 1999; Accepted, 16th March 1999
Paper E/9/01226E
C
1
2
3
4
5
6
7
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
53.6
32.1
24.6
40.1
38.2
22.8
29.5
18.3
29.1
76.4
19.1
16.1b
29.1
70.8
16.2
54.6
53.5
31.7b
24.5b
38.5
38.9
25.7c
19.4c
17.9b
29.1b
76.1
26.1
11.7
73.6
70.7
16.3
64.2
30.5b
24.5b
39.6
41.0
25.0c
25.6c
25.5b
33.1b
77.4
23.9
64.2
19.4
71.6
17.0
55.9
51.5
30.1b
32.0b
39.2
39.7
20.7c
21.1c
18.9b
35.1b
74.8
25.4
11.4
74.2
16.3
^
90.4
31.6
25.2
38.8
40.1
25.0
25.9
18.8
43.6
206.9
17.5
15.3
28.1
15.1
^
32.5
25.2
39.7
40.1
21.9
22.8
19.3
44.7
209.9
25.4
11.8
73.7
15.9
^
36.2b
30.5b
36.1
49.1
23.1c
27.2c
19.2b
40.5b
212.4
19.0
15.5
28.1
15.2
^
References
1
2
V. Lamare and R. Furstoss, Tetrahedron, 1990, 46, 4109.
R. Guillermo, J. R. Hanson and A. Truneh, J. Chem. Res. (S),
1997, 28.
3
4
5
6
7
8
A. San Feliciano, M. Medarde, M. Gordaliza, E. Del Olmo
and J. M. M. Del Corral, Phytochemistry, 1989, 28, 2717.
J. M. de Siqueira, C. D. de Oliveir and M. A. D. Boaventura,
Fitoterapia, 1997, LXVIII, 89.
D. P. de Lima, A. Beatriz, A. A. Ramos, J. M. de Siqueira, C.
C. de Oliveira and M. R. Marques, Quim Nova, 1997, 616.
H. J. M. Gijsen, J. B. P. A. Wijnberg, G. A. Stork and A. de
Groot, Tetrahedron, 1991, 48, 2465.
M. Miyazawa, T. Uemura and H. Kameoka, Phytochemistry,
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H. J. M. Gijsen, J. B. P. A. Wijnberg, C. van Ravenswaay and
A. de Groot, Tetrahedron, 1994, 50, 4733.
6
a The data were consistent with those reported in the literature,3
however, in this work a detailed assignment was accomplished.
b;cThe assignments for these signals within the same column may
be interchanged.