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Applied Microbiology and Biotechnology

, Volume 90, Issue 1, pp 147–157 | Cite as

Bioconversion of substituted naphthalenes and β-eudesmol with the cytochrome P450 BM3 variant F87V

  • Norihiko Misawa
  • Miho Nodate
  • Toshihiko Otomatsu
  • Keiko Shimizu
  • Chie Kaido
  • Miho Kikuta
  • Akira Ideno
  • Hiroshi Ikenaga
  • Jun Ogawa
  • Sakayu Shimizu
  • Kazutoshi Shindo
Biotechnologically Relevant Enzymes and Proteins
First Online:
Received:
Revised:
Accepted:

Abstract

Bioconversion of various substituted naphthalenes that contain 1-methoxy- and 1-ethoxy-naphthalenes, methylnaphthalenes, dimethylnaphthalenes, and naphthalenecarboxylic acid methyl esters were performed using recombinant Escherichia coli cells, which expressed the gene coding for a cytochrome P450 BM3 variant F87V (P450 BM3 (F87V)) that was N-terminally fused to an archaeal peptidyl–prolyl cis-trans isomerase. In addition, bioconversion experiments with the same substrates were carried out using those that expressed the phnA1A2A3A4 genes for a polycyclic aromatic hydrocarbon (PAH)-dihydroxylating dioxygenase, which originated from a PAH-utilizing marine bacterium Cycloclasticus sp. strain A5. Consequently, a variety of mono-hydroxylated derivatives were generated from these substituted naphthalenes. Oxidative aryl coupling was found to produce a novel compound 4,4′-diethoxy-[2,2′]-binaphthalenyl-1,1′-diol from 1-ethoxynaphthalene with the E. coli cells expressing the P450 BM3 (F87V) gene. This recombinant E. coli was further shown to introduce the hydroxyl group regio- and stereo-specifically into a sesquiterpene β-eudesmol.

Keywords

P450 BM3 Cytochrome P450 Peptidyl–prolyl cis-trans isomerase Dihydroxylating dioxygenase Naphthalene β-eudesmol 
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Notes

Acknowledgements

The authors thank Dr. Paul D. Fraser, Royal Holloway University of London, for his critical reading of this manuscript. This work was supported in part by the Research and Development Program for New Bio-industry Initiatives (2006–2010) from the Bio-oriented Technology Research Advancement Institution (BRAIN) of Japan. This work was also supported during MBI by New Energy and Industrial Technology Development Organization (NEDO) of Japan.

Supplementary material

253_2010_3064_MOESM1_ESM.doc (71 kb)
ESM 1 (DOC 71 kb)

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Norihiko Misawa
    • 1
    • 2
    • 6
  • Miho Nodate
    • 1
  • Toshihiko Otomatsu
    • 3
  • Keiko Shimizu
    • 1
  • Chie Kaido
    • 4
  • Miho Kikuta
    • 4
  • Akira Ideno
    • 1
  • Hiroshi Ikenaga
    • 1
  • Jun Ogawa
    • 5
  • Sakayu Shimizu
    • 5
  • Kazutoshi Shindo
    • 4
  1. 1.Marine Biotechnology Institute (MBI)KamaishiJapan
  2. 2.Central Laboratories for Frontier TechnologyKirin Holdings Co. Ltd., i-BIRDNonoichi-machiJapan
  3. 3.KNC Bio Research Center, KNC Laboratories Co., LtdKobeJapan
  4. 4.Department of Food and NutritionJapan Women’s UniversityTokyoJapan
  5. 5.Division of Applied Life Sciences, Graduate School of AgricultureKyoto UniversityKyotoJapan
  6. 6.Research Institute for Bioresources and BiotechnologyIshikawa Prefectural UniversityNonoichi-machiJapan

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