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Language: English

Conflict of Interest: In relation to this article, we declare that there is no conflict of interest.

Publication history: Received June 5, 2014
Accepted July 22, 2014

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/bync/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Ortho-hydroxylation of mammalian lignan enterodiol by cytochrome P450s from Actinomycetes sp.

EunOk Jung Kwon-Young Choi¹ Da-hye Jung Hyungdon Yun² Byung-Gee Kim^†

School of Chemical and Biological Engineering, Seoul National University, Seoul 137-070, Korea ¹Department of Environmental Engineering, Ajou University, Suwon, Gyeonggy 443-749, Korea ²Department of Bioscience & Biotechnology, Konkuk University, Seoul 143-701, Korea

Korean Journal of Chemical Engineering, March 2015, 32(3), 471-477(7), 10.1007/s11814-014-0211-3

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Abstract

An animal lignin, enterodiol (END), is known to be formed by conversion of secoisolariciresinol from flaxseed by intestinal bacteria. Thirteen bacteria strains were examined for their hydroxylation activity for END. Among them, Streptomyces avermitilis MA-4680 and Nocardia farcinica IFM10152 showed the highest hydroxylation activity for END. Reaction products profiled using GC/MS revealed that four products mono-hydroxylated in aliphatic position (Al-OH-END) and three products mono-hydroxylated in aromatic ring (Ar-OH-END) were found in S. avermitilis_x000D_ MA-4680, whereas only two Ar-OH-ENDs were detected in the case of N. farcinica IFM10152. From 15mg/L of END, 900 μg/L of Al-OH-END and 210 μg/L of 4-hydroxy END (4-OH-END) were produced by S. avermitilis MA-4680, and 300 μg/L of 2-hydroxy END (2-OH-END) and 480 μg/L of 4-OH-END were obtained by N. farcinica IFM10152. To find the P450s are responsible for the substrate specificity to END, 33 P450s from S. avermitilis MA-4680 and 26 P450s from N. farcinica IFM10152 were cloned and compared with coexpression of putidaredoxin reductase (camA)_x000D_ and putidaredoxin (camB) from Pseudomonas putida as redox partners in E. coli. As a result, Nfa45180 showed the highest hydroxylation activity especially for ortho-hydroxylation in aromatic ring in vivo. The results of the docking simulation of END into the homology model of Nfa45180 explained the reason for regio-specificity of the hydroxylation. To our knowledge, this is the first report of regioselective hydroxylation of END using microorganism P450s.

Keywords

Cytochrome P450 Enterodiol CYP154 GC/MS

References

Thompson LU, Baillieres Clin Endocrinol Metab., 12(4), 691 (1998)
McCann SE, Thompson LU, Nie J, Dorn J, Trevisan M, Shields PG, Ambrosone CB, Edge SB, Li HF, Kasprzak C, Freudenheim JL, Breast Cancer Res. Treat, 122(1), 229 (2010)
Murkies A, Dalais FS, Briganti EM, Burger HG, Healy DL, Wahlqvist ML, Davis SR, Menopause, 7(5), 289 (2000)
Meagher LP, Beecher GR, Flanagan VP, Li BW, J. Agric. Food Chem., 47(8), 3173 (1999)
Axelson M, Setchell KD, Febs Lett., 123(2), 337 (1981)
Jacobs E, Metzler M, J. Agric. Food Chem., 47, 1071 (1999)
Rufer CE, Kulling SE, J. Agric. Food Chem., 54, 2926 (2006)
Roh C, Seo SH, Choi KY, Cha M, Pandey BP, Kim JH, Park JS, Kim DH, Chang IS, Kim BG, J. Biosci. Bioeng., 108(1), 41 (2009)
Choi KY, Kim TJ, Koh SK, Roh CH, Pandey BP, Lee N, Kim BG, Biotechnol. J., 4(11), 1586 (2009)
Choi KY, Jung E, Jung DH, An BR, Pandey BP, Yun H, Sung C, Park HY, Kim BG, Microb. Cell. Fact., 11, 81 (2012)
Pandey BP, Lee N, Choi KY, Kim JN, Kim EJ, Kim BG, Appl. Microbiol. Biotechnol., 98(11), 5009 (2014)
Choi KY, Park HY, Kim BG, Enzyme Microb. Technol., 477, 327 (2010)
Thompson JD, Higgins DG, Gibson TJ, Nucleic Acids Res., 22(22), 4673 (1994)
Zhou H, Zhou Y, Bioinformatics, 21(18), 3615 (2005)
Sali A, Blundell TL, J. Mol. Biol., 234(3), 779 (1993)