JOURNAL OF CHEMICAL RESEARCH 2008 317
Table 1 13C NMR data determined in CDCl3 at 100 MHz of new
compounds 3, 11, 12 and 13
copper sulfate (0.1g) and magnesium sulfate (0.1g). The fungus was
grown for 2 days before the substrate (0.5g) in ethanol (30ml) was
distributed over 50 flasks. The fermentation was then continued for
further 7 days. The broth was filtered and the mycelium was washed
and the water layer extracted with ethyl acetate. The extracts were
washed with water and dried. The solvent was evaporated and the
residue was chromatographed on silica and eluted with an increasing
gradient of ethyl acetate in light petroleum.
3
11
12
13
C-1
36.8
30.9
68.3
41.7
67.3
62.2
76.4
37.5
45.1
34.5
21.6
31.4
49.4
50.9
21.8
35.8
220.2
14.1
17.1
—
30.9
21.7
51.9
55.8
45.9
26.7
31.7
35.4
52.6
34.8
20.3
36.9
42.3
50.4
23.7
27.9
82.4
12.2
13.6
21.0
171.2
31.5
24.8
71.0
76.1
43.6
24.8
30.9
35.2
51.2
35.9
20.3
37.1
42.7
51.1
23.2
27.5
81.6
12.2
14.3
22.1
170.8
37.2
38.1
211.6
43.6
45.3
27.1
30.3
34.2
49.9
35.3
20.9
35.9
47.1
51.6
35.0
53.1
64.1
13.8
16.9
—
C-2
C-3
C-4
17b-Acetoxy-3a,4a-epoxy-5a-androstane (3) (0.5g) gave 3a,4b-
dihydroxy-17b-acetoxy-5a-androstane (V)(35%) as white cubes
m.p.180–184°C. (Found: C,71.4; H, 9.6, C21 H34 O4 requires C,72.0;
C-5
C-6
C-7
H, 9.8%). FTIR: vmax/cm-1 3396, 1739. d (CDCl3, 400 MHz) 0.81
C-8
H
C-9
(3H, s, 18-H), 0.88 (3H, s, 19H), 2.05 (3H, s, 17b-OCOCH3), 3.62
(1H, br.s, 3b-H), 3.91 (1H, br.s, 4a-H).
C-10
C-11
C-12
C-13
C-14
C-15
C-16
C-17
C-18
C-19
17b-OCOCH3
17b-OCOCH3
3b-Hydroxy-5a,6a-epoxyandrostan-17-one (5) (0.5g) gave 3b,7a-
dihydroxy-5a,6a-epoxyandrostan-17-one (12) (21%) as white
needles. m.p.241–243°C. (Found C; 70.8, H; 8.3 C19H28O4 requires
C; 71.2, H; 8.8%). FTIR: vmax/cm-1 3390 and 1747. d (CDCl3,
H
400 MHz) 0.81 (3H, s, 18-H), 1.12 (3H, s, 19H), 3.31 (1H, d,
J = 3.76, 6b-H), 3.85 (IH, tt, J = 5.8 and J = 10.8, 3a-H). 4.05 (1H,
brs 7aH).
16a,17a-epoxy-5a-androstan-3b-ol (9) (0.5g) gave 16a,17a-
+
epoxy-5a-androstan-3-one (13) (12%) as an oil. M 288. Found
288.2081. C19H28O2 calculated 288.2089. FTIR: vmax/cm-1 1705.
—
d
(CDCl3, 400 MHz) 0.69 (3H, s, 18-H), 0.78 (3H, s, 19H), 3.06
(1H, d, J = 4 Hz, 17b-H), 3.31 (1H, d, J = 4 Hz, 16b-H).
H
stirred for 1 h. The TLC showed that all starting materials had
reacted. The solution was then washed with aqueous sodium sulfite,
aqueous sodium hydrogen carbonate, water and brine, and dried over
anhydrous sodium sulfate. The solvent was evaporated in vacuo to
give 17b-acetoxy-3a,4a-epoxy-5a-androstane (3) (0.82 g, 78%).
The product was crystallised from ethyl acetate as plates; m.p. 105–
107°C; (Found: C,75.8; H, 9.6, C21 H30 O3 requires C,75.9; H, 9.7%);
FTIR 1739 (C=O) cm-1.
The author thanks the Deanship of Scientific Research, King
Abdul Aziz University (KAU), Jeddah, Kingdom of Saudi
Arabia for financial support (Grant No 188/427).
Received 30 March 2008; accepted 30 April 2008
1H NMR (CDCl3, 400 MHz) 0.81 (3H, s, 18-H), 0.88 (3H, s, 19-H),
2.05 (3H, s, 17b-OCOCH3), 2.61 (1H, d, J = 4.4 Hz, 4b-H), 3.08 (1H,
m, 3b-H), 4.56 (1H, t, J = 8.5 Hz, 17a-H).
References
1
J.R. Hanson, An introduction to biotransformation in organic Chemistry,
W.H. Freeman, San Fancisco, 1998.
Biotransformation experiments
General fermentation details
2
2355.
3
4
5
6
A.P. Teske, Microbiology, 2005, 13, 402.
R.C. Cambie, P.S. Rutledge, D.W. Scott and P.D. Woodgate, Austral.
The fungus Mucor plumbeus (IMI 116688) was grown on shake
culture in 250 ml conical flasks on a medium (100ml) comprising
(per litre), Glucose (30g), potassium dihydrophosphate (1g),
magnesium sulfate (1g), ammonium tartrate (2g), yeast extract (1g),
calcium chloride (0.25g), sodium chloride (1g), ferrous ammonium
sulfate (1g), trace element solution 2 ml, distilled water to 1l,
neutralisation to pH 7 by adding NaOH.
7
N.E. Voishvillo, A.M. Turuta and A.V. Kamernitskii, Russ. Chem. Bull.,
1994, 43, 515.
8
9
J.R Hanson and K. Yildirim, J. Chem. Res. (S) 1999, 698.
Trace element solution contained (per litre) zinc sulfate (1.6g),
ferrous sulfate (1g), cobalt nitrate (1g), ammonium molybdate (1g),