Professional Documents
Culture Documents
Department of Pharmaceutical Sciences, Faculty of Pharmacy, Chiang Mai University, Chiang Mai, 50200, Thailand
Institute of Technology and Engineering, Massey University, Private Bag 11222, Palmerston North, 5320, New Zealand
*e-mail: pmpti006@chiangmai.ac.th
Received September 29, 2005
pounds other than hydrocortisone have been documented [18, 20, 24, 25].
Solid-state and submerged batch cultures of 22
molds were used to assess their capabilities for converting cortexolone to hydrocortisone. A color-reaction
assay was devised to rapidly establish if any cortexolone had been converted to hydrocortisone.
MATERIALS AND METHODS
Microorganisms. The following mycelial fungi
were used: two isolates of Cunninghamella blakesleeana; C. echinulata; Curvularia lunata ATCC 12017;
two isolates of Curvularia sp.; six isolates of Penicillium sp.; two isolates of Fusarium sp.; two isolates of
Aspergillus sp.; Calcarisporium antibioticum; Cladosporium sp.; Mucor sp.; Acremonium sp.; Paecilomyces sp.; and Rhizopus sp. The fungi were maintained on
Sabouraud dextrose agar (SDA; Difco, USA) slants at
4C.
Culture and biotransformation methodologies
All microorganisms were grown as monocultures. The
molds were subcultured on SDA Petri dishes at room
temperature 28 2C for 72120 h. A loopfull of the
culture was then transferred to 50 ml of enriched
growth medium held in a 250-ml shake flask (220 rpm).
The enriched medium contained the following components (g/l): glucose, 70; peptone, 1.0; yeast extract, 2.0;
L-asparagine, 0.7; nicotinic acid, 2 2 104; and riboflavin, 1.13 103. The pH was adjusted to 7.8 before
autoclaving 121C, 20 min. After 72 h of incubation,
the contents of the flask were divided into two identical
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MANOSROI et al.
CH2OH
O
CH3 OH
CH3
Cortexolone-containing SDA
Mycelial mat
H
O
Cortexolone
Petri disk
CH2OH
O
CH3 OH
HO
CH3
O
Hydrocortisone
Fig. 1. 11-Hydroxylation of cortexolone to hydrocortisone.
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Hydrocortisone yield and cortexolone remaining for different fungi and growth conditions*
Submerged culture method
Mold
residual
yield of
time at mearesidual
time at
maximum cortexolone, hydrocortisone, sured yield, cortexolone,
yield, d
% SD
% SD
days
% SD
yield of
hydrocortisone,
% SD
Cunninghamella blakesleeana
ATCC 8688 a
4.5 0.2
40.8 3.2
90.2 42.7
53.5 10.4
3.8 0.7
42.4 6.8
3.2 0.2
8.1 2.0
C. echinulata F 1307943
51.7 2.4
21.5 0.2
56.7 1.4
5.9 0.2
12.7 4.2
35.0 6.8
33.8 2.6
9.9 0.9
33.7 12.4
n.d.
56.2 14.9
n.d.
50.3 2.8
n.d.
4.0 1.4
n.d.
52.0 1.4
n.d.
14.0 0.0
n.d.
P. sp. F 613812
55.0 6.1
n.d.
48.7 0.8
n.d.
P.sp. F 70941
65.3 3.8
n.d.
0.3 0.1
n.d.
P. sp. F 879407
48.8 6.4
n.d.
20.2 11.1
n.d.
P. sp. F 879413
37.0 6.1
n.d.
29.3 10.4
n.d.
P. sp. F 104943
34.3 4.7
n.d.
20.3 3.8
n.d.
84.3 8.5
n.d.
34.8 33.7
n.d.
F. sp. F 127948
64.3 7.5
n.d.
41.5 2.1
n.d.
60.0 10.4
n.d.
43.2 1.2
n.d.
A. sp. F 879408
56.0 2.4
n.d.
29.5 5.0
n.d.
Calcarisporium antibioticum
40.0 1.9
n.d.
35.0 1.4
n.d.
48.0 9.9
n.d.
78.3 11.3
n.d.
42.7 0.9
n.d.
69.5 8.7
n.d.
120.7 12.3
n.d.
90.0 2.4
n.d.
102.3 11.8
n.d.
38.7 9.0
n.d.
81.3 13.2
n.d.
44.7 13.2
n.d.
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2006
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Vol. 42
No. 5
2006
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Vol. 42
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2006