Monooxygenation of Small Hydrocarbons Catalyzed by Bacterial Cytochrome P450s

  • Osami ShojiEmail author
  • Yoshihito Watanabe
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 851)


Cytochrome P450s (P450s) catalyze the NAD(P)H/O2-dependent monooxygenation of less reactive organic molecules under mild conditions. The catalytic activity of bacterial P450s is very high compared with P450s isolated from animals and plants, and the substrate specificity of bacterial P450s is also very high. Accordingly, their catalytic activities toward nonnative substrates are generally low especially toward small hydrocarbons. However, mutagenesis approaches have been very successful for engineering bacterial P450s for the hydroxylation of small hydrocarbons. On the other hand, “decoy” molecules, whose structures are very similar to natural substrates, can be used to trick the substrate recognition of bacterial P450s, allowing the P450s to catalyze oxidation reactions of nonnative substrates without any substitution of amino acid residues in the presence of decoy molecules. Thus, the hydroxylation of small hydrocarbons such as ethane, propane, butane and benzene can be catalyzed by P450BM3, a long-alkyl-chain hydroxylase, using substrate misrecognition of P450s induced by decoy molecules. Furthermore, a number of H2O2-dependent bacterial P450s can catalyze the peroxygenation of a variety of nonnative substrates through a simple substrate–misrecognition trick, in which catalytic activities and enantioselectivity are dependent on the structure of decoy molecules.


Bacterial P450s Mutagenesis approaches Gaseous alkanes Decoy molecules Hydrogen peroxide 



This work was supported, in part, by Grants-in-Aid for Scientific Research (S) to Y. W. (24225004) and for Young Scientists (A) to O. S. (21685018), and a Grant-in-Aid for Scientific Research on Innovative Areas “Molecular Activation Directed toward Straightforward Synthesis” to O. S. (25105724) from the Ministry of Education, Culture, Sports, Science and Technology (Japan).


  1. 1.
    Sono M, Roach MP, Coulter ED, Dawson JH (1996) Heme-containing oxygenases. Chem Rev 96:2841–2888CrossRefPubMedGoogle Scholar
  2. 2.
    Ortiz de Montellano PR (2005) Cytochrome P450: structure, mechanism, and biochemistry, 3rd edn. Kluwer Academic/Plenum Publishers, New YorkCrossRefGoogle Scholar
  3. 3.
    Denisov IG, Makris TM, Sligar SG, Schlichting I (2005) Structure and chemistry of cytochrome P450. Chem Rev 105:2253–2277CrossRefPubMedGoogle Scholar
  4. 4.
    Whitehouse CJC, Bell SG, Wong LL (2012) P450BM3 (CYP102A1): connecting the dots. Chem Soc Rev 41:1218–1260CrossRefPubMedGoogle Scholar
  5. 5.
    Fasan R (2012) Tuning P450 enzymes as oxidation catalysts. ACS Catal 2:647–666CrossRefGoogle Scholar
  6. 6.
    Dunford HB, Stillman JS (1976) Function and mechanism of action of peroxidases. Coord Chem Rev 19:187–251CrossRefGoogle Scholar
  7. 7.
    Rittle J, Green MT (2010) Cytochrome P450 compound I: capture, characterization, and C-H bond activation kinetics. Science 330:933–937CrossRefPubMedGoogle Scholar
  8. 8.
    Ost TWB, Clark J, Mowat CG, Miles CS, Walkinshaw MD, Reid GA, Chapman SK, Daff S (2003) Oxygen activation and electron transfer in flavocytochrome P450 BM3. J Am Chem Soc 125:15010–15020CrossRefPubMedGoogle Scholar
  9. 9.
    Sligar SG (1976) Coupling of spin, substrate, and redox equilibria in cytochrome P450. Biochemistry 15:5399–5406CrossRefPubMedGoogle Scholar
  10. 10.
    Murataliev MB, Feyereisen R (1996) Functional interactions in cytochrome P450BM3. Fatty acid substrate binding alters electron-transfer properties of the flavoprotein domain. Biochemistry 35:15029–15037CrossRefPubMedGoogle Scholar
  11. 11.
    Daff SN, Chapman SK, Turner KL, Holt RA, Govindaraj S, Poulos TL, Munro AW (1997) Redox control of the catalytic cycle of flavocytochrome P-450 BM3. Biochemistry 36:13816–13823CrossRefPubMedGoogle Scholar
  12. 12.
    Davydov R, Macdonald IDG, Makris TM, Sligar SG, Hoffman BM (1999) EPR and ENDOR of catalytic intermediates in cryoreduced native and mutant oxy-cytochromes P450cam: mutation-induced changes in the proton delivery system. J Am Chem Soc 121:10654–10655CrossRefGoogle Scholar
  13. 13.
    Fujishiro T, Shoji O, Nagano S, Sugimoto H, Shiro Y, Watanabe Y (2011) Crystal structure of H2O2-dependent cytochrome P450SPα with its bound fatty acid substrate. J Biol Chem 286:29941–29950CrossRefPubMedCentralPubMedGoogle Scholar
  14. 14.
    Lee DS, Yamada A, Sugimoto H, Matsunaga I, Ogura H, Ichihara K, Adachi S, Park SY, Shiro Y (2003) Substrate recognition and molecular mechanism of fatty acid hydroxylation by cytochrome P450 from Bacillus subtilis. Crystallographic, spectroscopic, and mutational studies. J Biol Chem 278:9761–9767CrossRefPubMedGoogle Scholar
  15. 15.
    Narhi LO, Fulco AJ (1986) Characterization of a catalytically self-sufficient 119,000-Dalton cytochrome P-450 monooxygenase induced by barbiturates in Bacillus megaterium. J Biol Chem 261:7160–7169PubMedGoogle Scholar
  16. 16.
    Boddupalli SS, Pramanik BC, Slaughter CA, Estabrook RW, Peterson JA (1992) Fatty acid monooxygenation by P450BM-3: product identification and proposed mechanisms for the sequential hydroxylation reactions. Arch Biochem Biophys 292:20–28CrossRefPubMedGoogle Scholar
  17. 17.
    Ravichandran KG, Boddupalli SS, Hasemann CA, Peterson JA, Deisenhofer J (1993) Crystal-structure of hemoprotein domain of P450BM-3, a prototype for microsomal P450s. Science 261:731–736CrossRefPubMedGoogle Scholar
  18. 18.
    Noble MA, Miles CS, Chapman SK, Lysek DA, Mackay AC, Reid GA, Hanzlik RP, Munro AW (1999) Roles of key active-site residues in flavocytochrome P450 BM3. Biochem J 339:371–379CrossRefPubMedCentralPubMedGoogle Scholar
  19. 19.
    Girvan HM, Marshall KR, Lawson RJ, Leys D, Joyce MG, Clarkson J, Smith WE, Cheesman MR, Munro AW (2004) Flavocytochrome P450 BM3 mutant A264E undergoes substrate-dependent formation of a novel heme iron ligand set. J Biol Chem 279:23274–23286CrossRefPubMedGoogle Scholar
  20. 20.
    Girvan HM, Toogood HS, Littleford RE, Seward HE, Smith WE, Ekanem IS, Leys D, Cheesman MR, Munro AW (2009) Novel haem co-ordination variants of flavocytochrome P450 BM3. Biochem J 417:65–76CrossRefPubMedGoogle Scholar
  21. 21.
    Li HY, Poulos TL (1997) The structure of the cytochrome P450BM-3 haem domain complexed with the fatty acid substrate, palmitoleic acid. Nat Struct Biol 4:140–146CrossRefPubMedGoogle Scholar
  22. 22.
    Fasan R, Chen MM, Crook NC, Arnold FH (2007) Engineered alkane-hydroxylating cytochrome P450(BM3) exhibiting nativelike catalytic properties. Angew Chem Int Ed 46:8414–8418CrossRefGoogle Scholar
  23. 23.
    Fasan R, Meharenna YT, Snow CD, Poulos TL, Arnold FH (2008) Evolutionary history of a specialized P450 propane monooxygenase. J Mol Biol 383:1069–1080CrossRefPubMedCentralPubMedGoogle Scholar
  24. 24.
    Chen MMY, Snow CD, Vizcarra CL, Mayo SL, Arnold FH (2012) Comparison of random mutagenesis and semi-rational designed libraries for improved cytochrome P450 BM3-catalyzed hydroxylation of small alkanes. Protein Eng Des Sel 25:171–178CrossRefPubMedGoogle Scholar
  25. 25.
    Meinhold P, Peters MW, Chen MMY, Takahashi K, Arnold FH (2005) Direct conversion of ethane to ethanol by engineered cytochrome P450BM3. ChemBioChem 6:1765–1768CrossRefPubMedGoogle Scholar
  26. 26.
    Staudt S, Müller CA, Marienhagen J, Böing C, Buchholz S, Schwaneberg U, Gröger H (2012) Biocatalytic hydroxylation of n-butane with in situ cofactor regeneration at low temperature and under normal pressure. Beilstein J Org Chem 8:186–191CrossRefPubMedCentralPubMedGoogle Scholar
  27. 27.
    Whitehouse CJC, Yang W, Yorke JA, Rowlatt BC, Strong AJF, Blanford CF, Bell SG, Bartlam M, Wong LL, Rao ZH (2010) Structural basis for the properties of two single-site proline mutants of CYP102A1 (P450BM3). ChemBioChem 11:2549–2556CrossRefPubMedGoogle Scholar
  28. 28.
    Glieder A, Farinas ET, Arnold FH (2002) Laboratory evolution of a soluble, self-sufficient, highly active alkane hydroxylase. Nat Biotechnol 20:1135–1139CrossRefPubMedGoogle Scholar
  29. 29.
    Xu F, Bell SG, Lednik J, Insley A, Rao ZH, Wong LL (2005) The heme monooxygenase cytochrome P450cam can be engineered to oxidize ethane to ethanol. Angew Chem Int Ed 44:4029–4032CrossRefGoogle Scholar
  30. 30.
    Bell SG, Stevenson JA, Boyd HD, Campbell S, Riddle AD, Orton EL, Wong LL (2002) Butane and propane oxidation by engineered cytochrome P450cam. Chem Commun 490–491Google Scholar
  31. 31.
    Farinas ET, Alcalde M, Arnold F (2004) Alkene epoxidation catalyzed by cytochrome P450 BM-3 139–3. Tetrahedron 60:525–528CrossRefGoogle Scholar
  32. 32.
    Dennig A, Lülsdorf N, Liu H, Schwaneberg U (2013) Regioselective o-hydroxylation of monosubstituted benzenes by P450 BM3. Angew Chem Int Ed 52:8459–8462CrossRefGoogle Scholar
  33. 33.
    Whitehouse CJC, Rees NH, Bell SG, Wong LL (2011) Dearomatisation of o-xylene by P450BM3 (CYP102A1). Chem Eur J 17:6862–6868CrossRefPubMedGoogle Scholar
  34. 34.
    Bell SG, Orton E, Boyd H, Stevenson JA, Riddle A, Campbell S, Wong LL (2003) Engineering cytochrome P450cam into an alkane hydroxylase. Dalton Trans 11:2133–2140CrossRefGoogle Scholar
  35. 35.
    Dennig A, Marienhagen J, Ruff AJ, Guddat L, Schwaneberg U (2012) Directed evolution of P450 BM3 into a p-xylene hydroxylase. ChemCatChem 4:771–773CrossRefGoogle Scholar
  36. 36.
    Kawakami N, Shoji O, Watanabe Y (2011) Use of perfluorocarboxylic acids to trick cytochrome P450BM3 into initiating the hydroxylation of gaseous alkanes. Angew Chem Int Ed 50:5315–5318CrossRefGoogle Scholar
  37. 37.
    Zilly FE, Acevedo JP, Augustyniak W, Deege A, Hausig UW, Reetz MT (2011) Tuning a P450 enzyme for methane oxidation. Angew Chem Int Ed 50:2720–2724CrossRefGoogle Scholar
  38. 38.
    Banks RE, Tatlow JC (1986) A guide to modern organofluorine chemistry. J Fluor Chem 33:227–346CrossRefGoogle Scholar
  39. 39.
    Kawakami N, Shoji O, Watanabe Y (2013) Direct hydroxylation of primary carbons in small alkanes by wild-type cytochrome P450BM3 containing perfluorocarboxylic acids as decoy molecules. Chem Sci 4:2344–2348CrossRefGoogle Scholar
  40. 40.
    Shoji O, Kunimatsu T, Kawakami N, Watanabe Y (2013) Highly selective hydroxylation of benzene to phenol by wild-type cytochrome P450BM3 assisted by decoy molecules. Angew Chem Int Ed 52:6606–6610CrossRefGoogle Scholar
  41. 41.
    Zhai PM, Wang LQ, Liu CH, Zhang SC (2005) Deactivation of zeolite catalysts for benzene oxidation to phenol. Chem Eng J 111:1–4CrossRefGoogle Scholar
  42. 42.
    Nordblom GD, White RE, Coon MJ (1976) Studies on hydroperoxide-dependent substrate hydroxylation by purified liver microsomal cytochrome P-450. Arch Biochem Biophys 175:524–533CrossRefPubMedGoogle Scholar
  43. 43.
    Renneberg R, Scheller F, Ruckpaul K, Pirrwitz J, Mohr P (1978) NADPD and H2O2-dependent reactions of cytochrome P-450LM compared with peroxidase catalysis. FEBS Lett 96:349–353CrossRefPubMedGoogle Scholar
  44. 44.
    Hrycay EG, Gustafsson JA, Ingelman–Sundberg M, Ernster L (1975) Sodium periodate, sodium chlorite, organic hydroperoxides, and H2O2 as hydroxylating agents in steroid hydroxylation reactions catalyzed by partially purified cytochrome P-450. Biochem Biophys Res Commun 66:209–216CrossRefPubMedGoogle Scholar
  45. 45.
    Renneberg R, Capdevila J, Chacos N, Estabrook RW, Prough RA (1981) Hydrogen peroxide-supported oxidation of benzo[a]pyrene by rat liver microsomal fractions. Biochem Pharmacol 30:843–848CrossRefPubMedGoogle Scholar
  46. 46.
    Holm KA, Engell RJ, Kupfer D (1985) Regioselectivity of hydroxylation of prostaglandins by liver microsomes supported by NADPH versus H2O2 in methylcholanthrene-treated and control rats: formation of novel prostaglandin metabolites. Arch Biochem Biophys 237:477–489CrossRefPubMedGoogle Scholar
  47. 47.
    Koo LS, Tschirret-Guth RA, Straub WE, Moenne-Loccoz P, Loehr TM, Ortiz de Montellano PR (2000) The active site of the thermophilic CYP119 from Sulfolobus solfataricus. J Biol Chem 275:14112–14123CrossRefPubMedGoogle Scholar
  48. 48.
    Ogura H, Nishida CR, Hoch UR, Perera R, Dawson JH, Ortiz de Montellano PR (2004) EpoK, a cytochrome P450 involved in biosynthesis of the anticancer agents epothilones A and B. Substrate-mediated rescue of a P450 enzyme. Biochemistry 43:14712–14721CrossRefPubMedGoogle Scholar
  49. 49.
    Anari MR, Josephy PD, Henry T, O’Brien PJ (1997) Hydrogen peroxide supports human and rat cytochrome P450 1A2-catalyzed 2-amino-3-methylimidazo[4,5-f]-quinoline bioactivation to mutagenic metabolites: significance of cytochrome P450 peroxygenase. Chem Res Toxicol 10:582–588CrossRefPubMedGoogle Scholar
  50. 50.
    Bui PH, Hankinson O (2009) Functional characterization of human cytochrome P450 2S1 using a synthetic gene-expressed protein in Escherichia coli. Mol Pharmacol 76:1031–1043CrossRefPubMedCentralPubMedGoogle Scholar
  51. 51.
    Niraula NP, Kanth BK, Sohng JK, Oh TJ (2011) Hydrogen peroxide-mediated dealkylation of 7-ethoxycoumarin by cytochrome P450 (CYP107AJ1) from Streptomyces peucetius ATCC27952. Enzym Microb Technol 48:181–186CrossRefGoogle Scholar
  52. 52.
    Goyal S, Banerjee S, Mazumdar S (2012) Oxygenation of monoenoic fatty acids by CYP175A1, an orphan cytochrome P450 from Thermus thermophilus HB27. Biochemistry 51:7880–7890CrossRefPubMedGoogle Scholar
  53. 53.
    Khan KK, He YA, He YQ, Halpert JR (2002) Site-directed mutagenesis of cytochrome P450eryF: implications for substrate oxidation, cooperativity, and topology of the active site. Chem Res Toxicol 15:843–853CrossRefPubMedGoogle Scholar
  54. 54.
    Matsumura H, Wakatabi M, Omi S, Ohtaki A, Nakamura N, Yohda M, Ohno H (2008) Modulation of redox potential and alteration in reactivity via the peroxide shunt pathway by mutation of cytochrome P450 around the proximal heme ligand. Biochemistry 47:4834–4842CrossRefPubMedGoogle Scholar
  55. 55.
    Zhang Z, Li Y, Stearns RA, Ortiz De Montellano PR, Baillie TA, Tang W (2002) Cytochrome P450 3A4-mediated oxidative conversion of a cyano to an amide group in the metabolism of pinacidil. Biochemistry 41:2712–2718CrossRefPubMedGoogle Scholar
  56. 56.
    Gelb MH, Heimbrook DC, Malkonen P, Sligar SG (1982) Stereochemistry and deuterium isotope effects in camphor hydroxylation by the cytochrome P450cam monooxygenase system. Biochemistry 21:370–377CrossRefPubMedGoogle Scholar
  57. 57.
    Poulos TL, Finzel BC, Howard AJ (1987) High-resolution crystal structure of cytochrome P450cam. J Mol Biol 195:687–700CrossRefPubMedGoogle Scholar
  58. 58.
    Yano JK, Wester MR, Schoch GA, Griffin KJ, Stout CD, Johnson EF (2004) The structure of human microsomal cytochrome P450 3A4 determined by X-ray crystallography to 2.05 Å resolution. J Biol Chem 279:38091–38094CrossRefPubMedGoogle Scholar
  59. 59.
    Gajhede M, Schuller DJ, Henriksen A, Smith AT, Poulos TL (1997) Crystal structure of horseradish peroxidase C at 2.15 Å resolution. Nat Struct Biol 4:1032–1038CrossRefPubMedGoogle Scholar
  60. 60.
    Sundaramoorthy M, Terner J, Poulos TL (1995) The crystal structure of chloroperoxidase: a heme peroxidase-cytochrome P450 functional hybrid. Structure 3:1367–1377CrossRefPubMedGoogle Scholar
  61. 61.
    Sundaramoorthy M, Terner J, Poulos TL (1998) Stereochemistry of the chloroperoxidase active site: crystallographic and molecular-modeling studies. Chem Biol 5:461–473CrossRefPubMedGoogle Scholar
  62. 62.
    Piontek K, Strittmatter E, Ullrich R, Grobe G, Pecyna MJ, Kluge M, Scheibner K, Hofrichter M, Plattner DA (2013) Structural basis of substrate conversion in a new aromatic peroxygenase: cytochrome P450 functionality with benefits. J Biol Chem 288:34767–34776CrossRefPubMedCentralPubMedGoogle Scholar
  63. 63.
    Wang XS, Peter S, Ullrich R, Hofrichter M, Groves JT (2013) Driving force for oxygen-atom transfer by heme-thiolate enzymes. Angew Chem Int Ed 52:9238–9241CrossRefGoogle Scholar
  64. 64.
    Wang XS, Peter S, Kinne M, Hofrichter M, Groves JT (2012) Detection and kinetic characterization of a highly reactive heme-thiolate peroxygenase compound I. J Am Chem Soc 134:12897–12900CrossRefPubMedCentralPubMedGoogle Scholar
  65. 65.
    Goodin DB, Mcree DE (1993) The Asp-His-iron triad of cytochrome c peroxidase controls the reduction potential, electronic structure, and coupling of the tryptophan free-radical to the heme. Biochemistry 32:3313–3324CrossRefPubMedGoogle Scholar
  66. 66.
    Yamada Y, Fujiwara T, Sato T, Igarashi N, Tanaka N (2002) The 2.0 Å crystal structure of catalase-peroxidase from Haloarcula marismortui. Nat Struct Biol 9:691–695CrossRefPubMedGoogle Scholar
  67. 67.
    Ko TP, Day J, Malkin AJ, McPherson A (1999) Structure of orthorhombic crystals of beef liver catalase. Acta Crystallogr D 55:1383–1394CrossRefPubMedGoogle Scholar
  68. 68.
    Watanabe Y, Ueno T (2003) Introduction of P450, peroxidase, and catalase activities into myoglobin by site-directed mutagenesis: diverse reactivities of compound I. Bull Chem Soc Jpn 76:1309–1322CrossRefGoogle Scholar
  69. 69.
    Ozaki SI, Roach MP, Matsui T, Watanabe Y (2001) Investigations of the roles of the distal heme environment and the proximal heme iron ligand in peroxide activation by heme enzymes via molecular engineering of myoglobin. Acc Chem Res 34:818–825CrossRefGoogle Scholar
  70. 70.
    Watanabe Y, Nakajima H, Ueno T (2007) Reactivities of oxo and peroxo intermediates studied by hemoprotein mutants. Acc Chem Res 40:554–562CrossRefPubMedGoogle Scholar
  71. 71.
    Li QS, Ogawa J, Schmid RD, Shimizu S (2005) Indole hydroxylation by bacterial cytochrome P450 BM-3 and modulation of activity by cumene hydroperoxide. Biosci Biotechnol Biochem 69:293–300CrossRefPubMedGoogle Scholar
  72. 72.
    Vidal-Limón A, Aguila S, Ayala M, Batista CV, Vazquez-Duhalt R (2013) Peroxidase activity stabilization of cytochrome P450BM3 by rational analysis of intramolecular electron transfer. J Inorg Biochem 122:18–26CrossRefPubMedGoogle Scholar
  73. 73.
    Joo H, Lin ZL, Arnold FH (1999) Laboratory evolution of peroxide-mediated cytochrome P450 hydroxylation. Nature 399:670–673CrossRefPubMedGoogle Scholar
  74. 74.
    Kumar S, Chen CS, Waxman DJ, Halpert JR (2005) Directed evolution of mammalian cytochrome P450 2B1: mutations outside of the active site enhance the metabolism of several substrates, including the anticancer prodrugs cyclophosphamide and ifosfamide. J Biol Chem 280:19569–19575CrossRefPubMedGoogle Scholar
  75. 75.
    Kumar S, Liu H, Halpert JR (2006) Engineering of cytochrome P450 3A4 for enhanced peroxide-mediated substrate oxidation using directed evolution and site-directed mutagenesis. Drug Metab Dispos 34:1958–1965CrossRefPubMedGoogle Scholar
  76. 76.
    Li QS, Ogawa J, Shimizu S (2001) Critical role of the residue size at position 87 in H2O2-dependent substrate hydroxylation activity and H2O2 inactivation of cytochrome P450BM-3. Biochem Biophys Res Commun 280:1258–1261CrossRefPubMedGoogle Scholar
  77. 77.
    Schwaneberg U, Schmidt-Dannert C, Schmitt J, Schmid RD (1999) A continuous spectrophotometric assay for P450 BM-3, a fatty acid hydroxylating enzyme, and its mutant F87A. Anal Biochem 269:359–366CrossRefPubMedGoogle Scholar
  78. 78.
    Cirino PC, Arnold FH (2002) Regioselectivity and activity of cytochrome P450 BM-3 and mutant F87A in reactions driven by hydrogen peroxide. Adv Synth Catal 344:932–937CrossRefGoogle Scholar
  79. 79.
    Cirino PC, Arnold FH (2003) A self-sufficient peroxide-driven hydroxylation biocatalyst. Angew Chem Int Ed 42:3299–3301CrossRefGoogle Scholar
  80. 80.
    Sanchez-Sanchez L, Roman R, Vazquez-Duhalt R (2012) Pesticide transformation by a variant of CYPBM3 with improved peroxygenase activity. Pestic Biochem Physiol 102:169–174CrossRefGoogle Scholar
  81. 81.
    Matsunaga I, Sumimoto T, Ueda A, Kusunose E, Ichihara K (2000) Fatty acid-specific, regiospecific, and stereospecific hydroxylation by cytochrome P450 (CYP152B1) from Sphingomonas paucimobilis: substrate structure required for α-hydroxylation. Lipids 35:365–371CrossRefPubMedGoogle Scholar
  82. 82.
    Imai Y, Matsunaga I, Kusunose E, Ichihara K (2000) Unique heme environment at the putative distal region of hydrogen peroxide-dependent fatty acid α-hydroxylase from Sphingomonas paucimobilis (peroxygenase P450SPα). J Biochem 128:189–194CrossRefPubMedGoogle Scholar
  83. 83.
    Matsunaga I, Yamada M, Kusunose E, Miki T, Ichihara K (1998) Further characterization of hydrogen peroxide-dependent fatty acid α-hydroxylase from Sphingomonas paucimobilis. J Biochem 124:105–110CrossRefPubMedGoogle Scholar
  84. 84.
    Matsunaga I, Sumimoto T, Kusunose E, Ichihara K (1998) Phytanic acid α-hydroxylation by bacterial cytochrome P450. Lipids 33:1213–1216CrossRefPubMedGoogle Scholar
  85. 85.
    Matsunaga I, Yokotani N, Gotoh O, Kusunose E, Yamada M, Ichihara K (1997) Molecular cloning and expression of fatty acid α-hydroxylase from Sphingomonas paucimobilis. J Biol Chem 272:23592–23596CrossRefPubMedGoogle Scholar
  86. 86.
    Matsunaga I, Yamada M, Kusunose E, Nishiuchi Y, Yano I, Ichihara K (1996) Direct involvement of hydrogen peroxide in bacterial α-hydroxylation of fatty acid. FEBS Lett 386:252–254CrossRefPubMedGoogle Scholar
  87. 87.
    Lee DS, Yamada A, Matsunaga I, Ichihara K, Adachi SI, Park SY, Shiro Y (2002) Crystallization and preliminary X-ray diffraction analysis of fatty-acid hydroxylase cytochrome P450BSβ from Bacillus subtilis. Acta Crystallogr D 58:687–689CrossRefPubMedGoogle Scholar
  88. 88.
    Matsunaga I, Shiro Y (2004) Peroxide-utilizing biocatalysts: structural and functional diversity of heme-containing enzymes. Curr Opin Chem Biol 8:127–132CrossRefPubMedGoogle Scholar
  89. 89.
    Matsunaga I, Sumimoto T, Ayata M, Ogura H (2002) Functional modulation of a peroxygenase cytochrome P450: novel insight into the mechanisms of peroxygenase and peroxidase enzymes. FEBS Lett 528:90–94CrossRefPubMedGoogle Scholar
  90. 90.
    Matsunaga I, Ueda A, Fujiwara N, Sumimoto T, Ichihara K (1999) Characterization of the ybdT gene product of Bacillus subtilis: novel fatty acid β-hydroxylating cytochrome P450. Lipids 34:841–846CrossRefPubMedGoogle Scholar
  91. 91.
    Matsunaga I, Ueda A, Sumimoto T, Ichihara K, Ayata M, Ogura H (2001) Site-directed mutagenesis of the putative distal helix of peroxygenase cytochrome P450. Arch Biochem Biophys 394:45–53CrossRefPubMedGoogle Scholar
  92. 92.
    Matsunaga I, Yamada A, Lee DS, Obayashi E, Fujiwara N, Kobayashi K, Ogura H, Shiro Y (2002) Enzymatic reaction of hydrogen peroxide-dependent peroxygenase cytochrome P450s: kinetic deuterium isotope effects and analyses by resonance Raman spectroscopy. Biochemistry 41:1886–1892CrossRefPubMedGoogle Scholar
  93. 93.
    Girhard M, Schuster S, Dietrich M, Durre P, Urlacher VB (2007) Cytochrome P450 monooxygenase from Clostridium acetobutylicum: a new α-fatty acid hydroxylase. Biochem Biophys Res Commun 362:114–119CrossRefPubMedGoogle Scholar
  94. 94.
    Rude MA, Baron TS, Brubaker S, Alibhai M, Del Cardayre SB, Schirmer A (2011) Terminal olefin (1-alkene) biosynthesis by a novel P450 fatty acid decarboxylase from Jeotgalicoccus species. Appl Environ Microbiol 77:1718–1727CrossRefPubMedCentralPubMedGoogle Scholar
  95. 95.
    Matsunaga I, Kusunose E, Yano I, Ichihara K (1994) Separation and partial characterization of soluble fatty acid α-hydroxylase from Sphingomonas paucimobilus. Biochem Biophys Res Commun 201:1554–1560CrossRefPubMedGoogle Scholar
  96. 96.
    Belcher J, McLean KJ, Matthews S, Woodward LS, Fisher K, Rigby SE, Nelson DR, Potts D, Baynham MT, Parker DA, Leys D, Munro AW (2014) Structure and biochemical properties of the alkene producing cytochrome P450 OleTJE (CYP152L1) from the Jeotgalicoccus sp. 8456 bacterium. J Biol Chem 289:6535–6550CrossRefPubMedCentralPubMedGoogle Scholar
  97. 97.
    Shoji O, Fujishiro T, Nakajima H, Kim M, Nagano S, Shiro Y, Watanabe Y (2007) Hydrogen peroxide dependent monooxygenations by tricking the substrate recognition of cytochrome P450BSβ. Angew Chem Int Ed 46:3656–3659CrossRefGoogle Scholar
  98. 98.
    Fujishiro T, Shoji O, Watanabe Y (2010) Non-covalent modification of the active site of cytochrome P450 for inverting the stereoselectivity of monooxygenation. Tetrahedron Lett 52:395–397CrossRefGoogle Scholar
  99. 99.
    Shoji O, Wiese C, Fujishiro T, Shirataki C, Wünsch B, Watanabe Y (2010) Aromatic C–H bond hydroxylation by P450 peroxygenases: a facile colorimetric assay for monooxygenation activities of enzymes based on Russig’s blue formation. J Biol Inorg Chem 15:1109–1115CrossRefPubMedGoogle Scholar
  100. 100.
    Fujishiro T, Shoji O, Kawakami N, Watanabe T, Sugimoto H, Shiro Y, Watanabe Y (2012) Chiral-substrate-assisted stereoselective epoxidation catalyzed by H2O2-dependent cytochrome P450SPα. Chem Asian J 7:2286–2293CrossRefPubMedGoogle Scholar
  101. 101.
    Shoji O, Fujishiro T, Nagano S, Tanaka S, Hirose T, Shiro Y, Watanabe Y (2010) Understanding substrate misrecognition of hydrogen peroxide dependent cytochrome P450 from Bacillus subtilis. J Biol Inorg Chem 15:1331–1339CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  1. 1.Bioinorganic Chemistry Laboratory, Department of ChemistryNagoya UniversityNagoyaJapan
  2. 2.Bioinorganic Chemistry Laboratory, Research Center for Materials ScienceNagoya UniversityNagoyaJapan

Personalised recommendations