Hydroxylation of Testosterone by Bacterial Cytochromes P450
311
C57BL/6J mice. Eur. J. Biochem., 57, 9–14 (1975).
introduced methine proton (ꢂH 4.02) showed a mutual
correlation. ꢁ-Orientation was decided by comparing the
melting point of this product (191.5–192.5 ꢀC) to those
in the literature (ꢁ: 192–193 ꢀC; ꢀ: 183.5–185.5 ꢀC).
11) Mahato, S. B., and Mukherjee, A., Microbial trans-
formation of testosterone by Aspergillus fumigatus. J.
Steroid Biochem., 21, 341–342 (1984).
12) Smith, K. E., Latif, S., and Kirk, D. N., Microbial
transformation of steroids—II. Transformations of pro-
gesterone, testosterone and androstenedione by Phyco-
myces blakesleeanus. J. Steroid Biochem., 32, 445–451
(1989).
13) Smith, K. E., Latif, S., and Kirk, D. N., Microbial
transformations of steroids—VI. Transformation of
testosterone and androsterone by Botryosphaerica
obtusa. J. Steroid Biochem., 35, 115–120 (1990).
Acknowledgments
We thank T. Dairi of Toyama Prefectural University,
F. Kato of Toho University, Y. Anzai of Toho
University, and H. Ikeda of Kitasato Institute for Life
Sciences for providing genes for actinomycete P450s.
We also thank T. Kaneko of the Kazusa DNA Research
Institute for providing type strains of B. japonicum and
M. loti.
´
14) Kolek, T., and Swizdor, A., Biotransformation XLV:
transformations of 4-ene-3-oxo steroids in Fusarium
culmorum culture. J. Steroid Biochem. Molec. Biol., 67,
63–69 (1998).
15) Arisawa, A., and Kumeda, A., World Patent WO2003/
087381 (Oct. 23, 2003).
References
1) Evans, W. E., and Relling, M. V., Pharmacogenomics:
translating function genomics into rational therapeutics.
Science, 286, 487–491 (1999).
2) Dutton, D. R., McMillen, S. K., Sonderfan, A. J.,
Thomas, P. E., and Parkinson, A., Studies on the rate-
determining factor in testosterone hydroxylation by rat
liver microsomal cytochrome P-450: evidence against
cytochrome P-450 isozyme: isozyme interaction. Arch.
Biochem. Biophys., 255, 316–328 (1987).
16) Julian, A. P., Matthew, C. L., and Bilal, A., Putidar-
edoxin reductase and putidaredoxin cloning, sequence
determination, and heterologous expression of the
proteins. J. Biol. Chem., 265, 6066–6073 (1990).
17) Ishikawa, J., Yamashita, A., Mikami, Y., Hoshino, Y.,
Kurita, H., Hotta, K., Shibata, T., and Hattori, M., The
complete genomic sequence of Nocardia farcinica IFM
10152. Proc. Natl. Acad. Sci. U.S.A., 101, 14925–14930
(2004).
3) Sonderfan, A. J., Arlotto, M. P., and Parkinson, A.,
Identification of the cytochrome P-450 isozymes respon-
sible for testosterone oxidation in rat lung, kidney, and
testis: evidence that cytochrome P-450a (P450IIAI) is
the physiologically important testosterone 7ꢁ-hydrox-
ylase in rat testis. Endocrinology, 125, 857–866 (1989).
4) Krauser, J. A., Voehler, M., Tseng, L., Shefer, A. B.,
Godejohann, M., and Guengerich, F. P., Testosterone
1ꢀ-hydroxylation by human cytochrome P450 3A4. Eur.
J. Biochem., 271, 3962–3969 (2004).
5) Gustafsson, J. A., and Lisboa, B. P., Biosynthesis of 6-
beta-hydroxytestosterone from testosterone by human
fetal liver microsomes. Steroids, 11, 555–563 (1968).
6) Lisboa, B. P., and Gustafsson, J. A., Studies on the
metabolism of steroids in the foetus: biosynthesis of 6ꢁ-
hydroxytestosterone in the human foetal liver. Biochem.
J., 115, 583–586 (1969).
18) Dairi, T., Hamano, Y., Kuzuyama, T., Itoh, N., Furihata,
K., and Seto, H., Eubacterial diterpene cyclase genes
essential for production of the isoprenoid antibiotic
terpentecin. J. Bacteriol., 183, 6085–6094 (2001).
19) Isshiki, K., Tamamura, T., Sawa, T., Naganawa, H.,
Takeuchi, T., and Umezawa, H., Biosynthetic studies of
terpentecin. J. Antibiot., 39, 1634–1635 (1986).
20) Inoue, M., Takada, Y., Muto, N., Beppu, T., and
Horinouchi, S., Characterization and expression of a
P450-like mycinamicin biosynthesis gene using a novel
Micromonospora-Escherichia coli shuttle cosmid vector.
Mol. Gen. Genet., 245, 456–464 (1994).
21) Rodriguez, A. M., Olano, C., Mendez, C., Hutchinson,
C. R., and Salas, J. A., A cytochrome P450-like gene
possibly involved in oleandomycin biosynthesis by
Streptomyces antibioticus. FEMS Microbiol. Lett., 127,
117–120 (1995).
7) Cheng, K. C., and Schenkman, J. B., Testosterone
metabolism by cytochrome P-450 isozymes RLM3 and
RLM5 by microsomes. J. Biol. Chem., 258, 11738–
11744 (1983).
8) Wood, A. W., Swinney, D. C., Thomas, P. E., Ryan,
D. E., Hall, P. F., Levin, W., and Garland, W. A.,
Mechanism of androstenedione formation from testos-
terone and epitestosterone catalyzed by purified cyto-
chrome P-450b. J. Biol. Chem., 263, 17322–17332
(1988).
9) Jansson, I., Mole, J., and Schenkman, J. B., Purification
and characterization of a new form (RLM2) of liver
microsomal cytochrome P-450 from untreated rat. J.
Biol. Chem., 260, 7084–7093 (1985).
10) Ford, H. C., Wheeler, R., and Engel, L. L., Hydrox-
ylation of testosterone at carbons 1, 2, 6, 7, 15 and 16 by
the hepatic microsomal fraction from adult female
22) Rauschenbach, R., Isernhagen, M., Noeske-Jungblut, C.,
Boidol, W., and Siewert, G., Cloning, sequencing and
expression of the gene for cytochrome P450meg, the
steroid-15 beta-monooxygenase from Bacillus megate-
rium ATCC 13368. Mol. Gen. Genet., 241, 170–176
(1993).
23) Edwards, R. J., Murray, B. P., Singleton, A. M., and
Boobis, A. R., Orientation of cytochrome P450 in the
endoplasmic reticulum. Biochemistry, 30, 71–76 (1991).
24) Gotoh, O., Evolution and differentiation of P-450
genes. In ‘‘Cytochrome P-450’’ 2nd ed., eds. Omura,
T., Ishimura, Y., and Fujii-Kuriyama, Y., Kodansha,
Tokyo/VCH, Weinheim, pp. 255–272 (1993).
25) Ohta, K., Agematu, H., Yamada, T., Kaneko, K., and
Tsuchida, T., Production of human metabolites of
cyclosporin A, AM1, AM4N and AM9, by microbial
conversion. J. Biosci. Bioeng., 99, 390–395 (2005).