Skip to main content
SpringerLink
Log in

Phytanic acid α-hydroxylation by bacterial cytochrome P450

  • Published:
Lipids

Abstract

Fatty acid α-hydroxylase, a cytochrome P450 enzyme, from Sphingomonas paucimobilis, utilizes various straight-chain fatty acids as substrates. We investigated whether a recombinant fatty acid α-hydroxylase is able to metabolize phytanic acid, a methyl-branched fatty acid. When phytanic acid was incubated with the recombinant enzyme in the presence of H2O2, a reaction product was detected by gas chromatography, whereas a reaction product was not detected in the absence of H2O2. When a heat-inactivated enzyme was used, a reaction product was not detected with any concentration of H2O2. Analysis of the methylated product by gas chromatography-mass spectrometry revealed a fragmentation pattern of 2-hydroxyphytanic acid methyl ester. By single-ion monitoring, the mass ion and the characteristic fragmentation ions of 2-hydroxyphytanic acid methyl ester were detected at the retention time corresponding to the time of the product observed on the gas chromatogram. The K m value for phytanic acid was approximately 50 μM, which was similar to that for myristic acid, although the calculated V max for phytanic acid was about 15-fold lower than that for myristic acid. These results indicate that a bacterial cytochrome P450 is able to oxidize phytanic acid to form 2-hydroxyphytanic acid.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

FAAH:

fatty acid α-hydroxylase

GC-MS:

gas chromatography-mass spectrometry

P450:

cytochrome P450

References

  1. Yabuuchi, E., Yano, I., Ovaizu, H., Hashimoto, Y., Ezaki, T., and Yamamoto, H. (1990) Proposals of Sphingomonas paucimobilis gen. nov. and comb., Sphingomonas parapaucimobilis sp. nov., Sphingomonas yanoikuyae sp. nov., Sphingomonas adhaesiva sp. nov., Sphingomonas capsulata comb. nov., and two Genospecies of the Genus Sphingomonas. Microbiol. Immunol. 34, 99–119.

    PubMed  CAS  Google Scholar 

  2. Matsunaga, I., Yamada, M., Kusunose, E., Miki, T., and Ichihara, K. (1998) Further Characterization of Fatty Acid α-Hydroxylase from Sphingomonas paucimobilis, J. Biochem. 124, 105–110.

    PubMed  CAS  Google Scholar 

  3. Matsunaga, I., Yamada, M., Kusunose, E., Nishiuchi, Y., Yano, I., and Ichihara, K. (1996) Direct Involvement of Hydrogen Peroxide in Bacterial α-Hydroxylation of Fatty Acid, FEBS Lett. 386, 252–254.

    Article  PubMed  CAS  Google Scholar 

  4. Matsunaga, I., Yokotani, N., Gotoh, O., Kusunose, E., Yamada, M., and Ichihara, K. (1997) Molecular Cloning and Expression of Fatty Acid α-Hydroxylase from Sphingomonas paucimobilis, J. Biol. Chem. 272, 23592–23596.

    Article  PubMed  CAS  Google Scholar 

  5. Gotoh, O. (1993) Evolution and Differentiation of P450 Genes, in Cytochrome P450 (Omura, T., Ishimura, Y., and Fujii-Kuriyama, Y., eds.) 2nd edn., pp. 255–272, Kodansha, Tokyo.

    Google Scholar 

  6. Narhi, L.O., and Fulco, A.J. (1986) Characterization of a Catalytically Self-Sufficient 119,000-Dalton Cytochrome P-450 Monooxygenase Induced by Barbiturate in Bacillus megaterium, J. Biol. Chem. 261, 7160–7169.

    PubMed  CAS  Google Scholar 

  7. English, N., Palmer, C.N.A., Alworth, W.L., Kang, L., Hughes, V., and Wolf, C.R. (1997) Fatty Acid Signals in Bacillus megaterium Are Attenuated by Cytochrome P-450-Mediated Hydroxylation, Biochem. J. 327, 363–368.

    PubMed  CAS  Google Scholar 

  8. Tsai, S.-C., Avigan, J., and Steinberg, D. (1969) Studies on the α-Oxidation of Phytanic Acid by Rat Liver Mitochondria, J. Biol. Chem. 244, 2682–2692.

    PubMed  CAS  Google Scholar 

  9. Steinberg, D. (1989) Refsum Disease, in The Metabolic Basis of Inherited Disease (Scriver, C.R., Beaudet, A.L., Sly, W.S., and Valle, D., eds.) 6th edn., pp. 1533–1550. McGraw-Hill, New York.

    Google Scholar 

  10. Skjeldal, O.H., and Stokke, O. (1988) Evidence Against α-Hydroxyphytanic Acid as an Intermediate in the Metabolism of Phytanic Acid, Scand. J. Clin. Lab. Invest. 48, 97–102.

    PubMed  CAS  Google Scholar 

  11. Sawamura, A., Kusunose, E., Satouchi, K., and Kusunose, M. (1993) Catalytic Properties of Rabbit Kidney Fatty Acid ω-Hydroxylase Cytochrome P-450ka2 (CYP 4A7), Biochim. Biophys. Acta 1168, 30–36.

    PubMed  CAS  Google Scholar 

  12. Mihalik, S.J., Morrell, J.C., Kim, D., Sacksteder, K.A., Watkins, P.A., and Gould, S.J. (1997) Identification of PAHX, a Refsum Disease Gene, Nature Genet. 17, 185–189.

    Article  PubMed  CAS  Google Scholar 

  13. Jansen, G.A., Ofman, R., Ferdinandusse, S., Ijlst, L., Muijsers, A.O., Skjeldal, O.H., Stokke, O., Jakobs, C., Besley, G.T.N., Wraith, E.J., and Wanders, R.J.A. (1997) Refsum Disease Is Caused by Mutation in the Phytanoyl-CoA Hydroxylase Gene, Nature Genet 17, 190–193.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Osaka Prefectural Institute of Public Health, 537-0025, Osaka, Japan

    Tatsuo Sumimoto

  2. Department of Molecular Regulation, Osaka City University Medical School, 1-4-3 Asahi-machi, Abeno-ku, 545-8585, Osaka, Japan

    Isamu Matsunaga, Tatsuo Sumimoto, Emi Kusunose & Kosuke Ichihara

Authors
  1. Isamu Matsunaga
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tatsuo Sumimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Emi Kusunose
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kosuke Ichihara
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Isamu Matsunaga.

About this article

Cite this article

Matsunaga, I., Sumimoto, T., Kusunose, E. et al. Phytanic acid α-hydroxylation by bacterial cytochrome P450. Lipids 33, 1213–1216 (1998). https://doi.org/10.1007/s11745-998-0325-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11745-998-0325-4

Keywords

Search

Navigation