TY - JOUR
T1 - Demonstration of a cytochrome P-450-dependent steroid 15β-hydroxylase in Bacillus megaterium
AU - Berg, Anders
AU - Carlström, Kjell
AU - Gustafsson, Jan Åke
AU - Ingelman-Sundberg, Magnus
N1 - Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.
PY - 1975/10/27
Y1 - 1975/10/27
N2 - The steroid 15β-hydroxylase system of Bacillus megaterium was obtained in a cell-free preparation through sonication. The strictly NADPH-dependent 15β-hydroxylase activity, measured using progesterone as substrate, was inhibited by carbon monoxide, SKF 525-A, imidazole and metyrapone, indicating that the reaction is cytochrome P-450-dependent. A 40-fold purification of cytochrome P-450 in cell-free extracts was obtained by chromatography on DEAE-cellulose yielding a concentration of 0.32 nmoles of cytochrome P-450 per mg of protein. This partially purified cytochrome P-450 preparation catalyzed 15β- and 15α-hydroxylation of progesterone in the presence of NaIO4 or NaClO2 but not in the presence of NADPH or NADH.
AB - The steroid 15β-hydroxylase system of Bacillus megaterium was obtained in a cell-free preparation through sonication. The strictly NADPH-dependent 15β-hydroxylase activity, measured using progesterone as substrate, was inhibited by carbon monoxide, SKF 525-A, imidazole and metyrapone, indicating that the reaction is cytochrome P-450-dependent. A 40-fold purification of cytochrome P-450 in cell-free extracts was obtained by chromatography on DEAE-cellulose yielding a concentration of 0.32 nmoles of cytochrome P-450 per mg of protein. This partially purified cytochrome P-450 preparation catalyzed 15β- and 15α-hydroxylation of progesterone in the presence of NaIO4 or NaClO2 but not in the presence of NADPH or NADH.
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U2 - 10.1016/0006-291X(75)90517-3
DO - 10.1016/0006-291X(75)90517-3
M3 - Article
C2 - 811222
AN - SCOPUS:0016777277
VL - 66
SP - 1414
EP - 1423
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 4
ER -