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Electron paramagnetic resonance analyses of biotransformation reactions with cytochrome P-450 immobilized on mesoporous molecular sieves

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Abstract

Mobil Crystalline Material (MCM-41) can be used for the immobilization of enzymes and the investigation of electron transfer in biological systems. Electron transfer between MCM-41 with aluminum (Al-MCM-41) and cytochrome P-450 (CYP2B4) was observed using electron paramagnetic resonance (EPR). When CYP2B4 was immobilized by adsorption, it catalyzed the conversion of aniline to p-aminophenol. The electron transfer was evidenced when the signal with a g value (also called g-factor or spectroscopic manifestation of the magnetic moment) of 1.98 increased at the same time that the signal with a g value 2.24 decreased due to the addition of NADPH to CYP2B4 immobilized on Al-MCM-41, indicating that FeIII was reduced to FeII. Therefore, it is possible that Al-MCM-41 participates in the electron transfer process in biological systems.

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References

  • Díaz JF, Balkus Jr KJ (1996) Enzyme immobilization in MCM-41 molecular sieve. J Mol Catal B: Enzym. 2:115–126

    Google Scholar 

  • Guengerich FP, Johnson WW (1997) Kinetics of ferric cytochrome P-450 reduction by NADPH cytochrome P-450 reductase. Rapid reduction in absence of substrate an variation among cytochrome P-450 system. Biochemistry 36:14741–14750

    Article  PubMed  CAS  Google Scholar 

  • Jeevarajan AS, Kispert LD, Piekara-Sady L (1993) An ENDOR study of carotenoid cation radicals on silica alumina. Chem Phys Lett 209:269–274

    Article  CAS  Google Scholar 

  • Kanaeva IP, Dedinskii IR, Skotselyas ED, Krainev AG, Guleva IV, Sevryukova IF, Koen YM, Kuznetsova GP, Bachmanova GI, Archakov AI. (1992) Comparative study of Monomeric Reconstituted and Membrane Microsomal Monooxygenase systems of the rabbit Liver. Arch Biochem Biophys 298:395–402

    Article  PubMed  CAS  Google Scholar 

  • Kittel C (1996) Introduction to solid state physics. John Wiley, New York, pp 540–553

  • Konovalova TA, Kispert LD (1998) EPR and ENDOR studies of carotenoid solid Lewis acid interactions. J Chem Soc Faraday Trans 94:1465–1468

    Article  CAS  Google Scholar 

  • Kresge CT, Leonowicz ME, Roth WJ, Vartuli JC, Beck JS (1992) Ordered mesoporous molecular sieves synthesized by a liquid-crystal template mechanism. Nature 359:710–712

    Article  CAS  Google Scholar 

  • LeLean JE, Moon N, Dunham WR, Coon MJ (2000) EPR spectrometry of cytochrome P-450 2B4: effects of mutations and substrate binding. Biochem Biophys Res Comm 276:762–766

    Article  PubMed  CAS  Google Scholar 

  • Lu AYH, Coon MJ (1968) Role of hemoprotein P-450 in fatty acid ω-hydroxylation in a soluble system from liver microsomes. J Biol Chem 243:1331–1332

    PubMed  CAS  Google Scholar 

  • Morgan ET, Coon MJ (1984) Effects of cytochrome b5 on cytochrome P-450 catalyzed reactions. Studies with manganese-substituted cytochrome b5. Drug Metab Dispos 12:358–364

    PubMed  CAS  Google Scholar 

  • Peisach J, Blumberg WE (1970) Electron paramagnetic resonance study of the high and low spin forms of cytochrome P-450 in liver and liver microsomes from a methylcholanthrene treated rabbit. Proc Natl Acad Sci 67:172–179

    Article  PubMed  CAS  Google Scholar 

  • Prakash AM, Sung-Suh HM, Kevan L (1998) Electron spin resonance evidence for isomorphous substitution of Titanium into titanosilicate Ti-MCM41 mesoporous molecular sieve. J Phys Chem B 102:857–864

    Article  CAS  Google Scholar 

  • Rein H, Ristau O, Friedrich J, Jaing GR, Ruckpaul K (1977) Evidence for the existence of a high spin-low spin equilibrium in liver microsomal cytochrome P-450. FEBS Lett 75:19–22

    Article  PubMed  CAS  Google Scholar 

  • Rosales MC, Mendieta JE, Vazquez JI, Miranda R, García LA, Trujillo FJ (2005) Immobilization of cytochrome P-450 on MCM-41 with different silicon/aluminum ratios. Microporous Mesoporous Mater 80:25–31

    Article  Google Scholar 

  • Seurioukova IF, Li H, Zhang H, Peterson JA, Poulos TL (1999) Structure of a cytochrome P-450 redox partner electron transfer complex. Proc Natl Acad Sci USA 96:863–1868

    Article  Google Scholar 

  • Shen S, Strobel HW (1992) The role of cytochrome P450 lysine residues in the interaction between cytochrome P450IA1 and NADPH cytochrome P450 reductase. Arch Biochem Biophys 294:83–90

    Article  PubMed  CAS  Google Scholar 

  • Tamburini PP, Schenkman JB (1986) Differences in the mechanism of funtional interaction between NADPH-cytochrome P-450 reductase and its redox partners. Mol Pharmacol 30:178–185

    PubMed  CAS  Google Scholar 

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Acknowledgements

The authors thank CONACYT and COFAA-SIP/IPN for financial support and the Chemical Sciences, Geosciences and Biosciences Division, Office of Basic Energy Sciences, US Department of Energy for operational support at the University of Alabama and the National Science Foundation for EPR Instrument Grants CHE-0342921 and CHE-0079498 at the same place. We thank Ignacio Colin and Rogelio Ruiz for technical support in the EPR measurements.

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Authors and Affiliations

  1. Sección de Estudios de Posgrado e Investigación y Departamento de Bioquímica de la, Escuela Superior de Medicina del Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón s/n, 11340, Mexico, D.F., Mexico

    Martha Rosales-Hernández & Jose Trujillo-Ferrara

  2. Department of Chemistry, University of Alabama, Box 870336, Tuscaloosa, AL, 35487-0336, USA

    Lowell Kispert

  3. Instituto Mexicano del Petróleo, Eje Central L. Cárdenas 152, 07730, Mexico, D.F., Mexico

    Eduardo Torres-Ramírez

  4. Departamento de Física, ESFM, IPN, Ave. Instituto Politécnico Nacional S/N, Edif. 9 U.P. Zacatenco, Col. San pedro Zacatenco, Mexico, D.F., 07738, Mexico

    Daniel Ramírez-Rosales & Rafael Zamorano-Ulloa

Authors
  1. Martha Rosales-Hernández
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  2. Lowell Kispert
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  3. Eduardo Torres-Ramírez
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  4. Daniel Ramírez-Rosales
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  5. Rafael Zamorano-Ulloa
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  6. Jose Trujillo-Ferrara
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Corresponding author

Correspondence to Jose Trujillo-Ferrara.

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Rosales-Hernández, M., Kispert, L., Torres-Ramírez, E. et al. Electron paramagnetic resonance analyses of biotransformation reactions with cytochrome P-450 immobilized on mesoporous molecular sieves. Biotechnol Lett 29, 919–924 (2007). https://doi.org/10.1007/s10529-007-9328-3

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  • DOI: https://doi.org/10.1007/s10529-007-9328-3

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