본문 바로가기
KERD
논문KCI 등재

In-silico and In-vitro based studies of Streptomyces peucetius CYP107N3 for oleic acid epoxidation

2

조회

0

다운로드

0

인용

0

참고문헌

키워드

주제분류화학

초록

Certain members of the cytochromes P450 superfamily metabolize polyunsaturated long-chain fatty acids to several classes of oxygenated metabolites. An approach based on in silico analysis predicted that Streptomyces peucetius CYP107N3might be a fatty acid-metabolizing enzyme, showing high homology with epoxidase enzymes. Homology modeling and docking studies of CYP107N3 showed that oleic acid can fit directly into the active site pocket of the double bond of oleic acid within optimum distance of 4.6 Å from the Fe. In order to confirm the epoxidation activity proposed by in silico analysis,a gene coding CYP107N3 was expressed in Escherichia coli. The purified CYP107N3 was shown to catalyze C9-C10epoxidation of oleic acid in vitro to 9,10-epoxy stearic acid confirmed by ESI-MS, HPLC-MS and GC-MS spectral analysis. [BMB Reports 2012; 45(12): 736-741]

Abstract

Certain members of the cytochromes P450 superfamily metabolize polyunsaturated long-chain fatty acids to several classes of oxygenated metabolites. An approach based on in silico analysis predicted that Streptomyces peucetius CYP107N3might be a fatty acid-metabolizing enzyme, showing high homology with epoxidase enzymes. Homology modeling and docking studies of CYP107N3 showed that oleic acid can fit directly into the active site pocket of the double bond of oleic acid within optimum distance of 4.6 Å from the Fe. In order to confirm the epoxidation activity proposed by in silico analysis,a gene coding CYP107N3 was expressed in Escherichia coli. The purified CYP107N3 was shown to catalyze C9-C10epoxidation of oleic acid in vitro to 9,10-epoxy stearic acid confirmed by ESI-MS, HPLC-MS and GC-MS spectral analysis. [BMB Reports 2012; 45(12): 736-741]

참고문헌 (39)

  1. Werck-Reichhart, D., "Cytochromes P450: a success story", Genome Biol., 1, pp.3003.1~3003.9 (2000)
  2. Funk, C. D., "Prostaglandins and leukotrienes: advances in eicosanoid biology", Science, 294, pp.1871~1875 (2001)
  3. Blee, E., "Impact of phyto-oxylipins in plant defense", Trends Plant Sci., 7, pp.315~322 (2002)
  4. Noverr, M. C., "Production of eicosanoids and other oxylipins bypathogenic eukaryotic microbes", Clin. Microbiol. Rev., 16, pp.517~533 (2003)
  5. Fer, M., "Determination of polyunsaturated fatty acid monoepoxides by high performance liquid chromatography- mass spectrometry", J. Chromatogr., A 1115, pp.1~7 (2006)
  6. Lauterbach, B., "Cytochrome P450-dependent eicosapentaenoic acid metabolites are novel BK channel activators", Hypertension, 39, pp.609~613 (2002)
  7. Ye, D., "Cytochrome p-450 epoxygenase metabolites of docosahexaenoate potently dilate coronary arterioles by activating large-conductance calcium-activated potassium channels", J. Pharmacol. Exp. Ther., 303, pp.768~776 (2002)
  8. Hercule, H. C., "The vasodilator 17,18-epoxyeicosatetraenoic acid targets the pore-forming BK alpha channel subunit in rodents", Exp. Physiol., 92, pp.1067~1076 (2007)
  9. Morin, C., "Relaxing effects of 17(18)-EpETE on arterial and airway smooth muscles in human lung", Am. J. Physiol. Lung Cell Mol. Physiol., 296, pp.L130~L139 (2009)
  10. Van Rollins, M., "Epoxygenase metabolites of docosahexaenoic and eicosapentaenoic acids inhibit platelet aggregation at concentrations below those affecting thromboxane synthesis", J. Pharmacol. Exp. Ther., 274, pp.798~804 (1995)
  11. Graham-Lorence, S., "An active site substitution, F87V, converts cytochrome P450 BM-3 into a regio- and stereoselective (14S,15R)-arachidonic acid epoxygenase", J. Biol. Chem., 272, pp.1127~1135 (1997)
  12. Roman, R. J., "P-450 metabolites of arachidonic acid in the control of cardiovascular function", Physiol. Rev., 82, pp.131~185 (2002)
  13. Fleming, I., "Epoxyeicosatrienoic acids, cell signaling and angiogenesis", Prostaglandins Other Lipid Mediat., 82, pp.60~67 (2007)
  14. Zeldin, D. C., "Biochemical characterization of the human liver cytochrome P450 arachidonic acid epoxygenase pathway", Arch. Biochem. Biophys., 330, pp.87~96 (1996)
  15. King, L. M., "Cloning of CYP2J2 gene and identification of polymorphisms", Mol. Pharmacol., 61, pp.840~852 (2002)
  16. Stark, K., "Oxygenation of polyunsaturated long chain fatty acids by recombinant CYP4F8 and CYP4F12 and catalytic importance of Tyr-125 and Gly-328 of CYP4F8", Arch. Biochem. Biophys., 441, pp.174~181 (2005)
  17. Shah, S., "Cloning, characterization and heterologous expression of polyketide synthase and P-450 oxidase involved in the biosynthesis of the antibiotic oleandomycin", J. Antibiot., 53, pp.502~508 (2000)
  18. Ward, S. L., "Chalcomycin biosynthesis gene cluster from Streptomyces bikiniensis: novel features of an unusual ketolide produced through expression of the chm polyketide synthase in Streptomyces fradiae", Antimicrob. Agent Chemother., 48, pp.4703~4712 (2004)
  19. Sali, A., "Evaluation of comparative protein modeling by MODELLER", Proteins, 23, pp.318~326 (1995)
  20. Venkatachalam, C. M., "LigandFit: a novel method for the shape-directed rapid docking of ligands to protein active sites", J. Mol. Graph. Model, 21, pp.289~307 (2003)
  21. Schenkman, J. B., "Substrate interaction with cytochrome P-450", Pharmacol. Ther., 12, pp.43~71 (1981)
  22. Kuo, T. M., "Factors influencing the production of novel compound, 7,10-dihydroxy-8(e)- octadecanoic acid, by pseudomonas aeruginosa PR3 (NRRL B-18602) in batch cultures", Curr. Microbiol., 47, pp.186~197 (2003)
  23. Kuo, T. M., "Production of 10-ketostearic acid and 10-hydroxystearicacid by strains of Sphingobacterium thalophilum isolatedfrom composted manure", Curr. Microbiol., 40, pp.105~109 (2000)
  24. Kuo, T. M., "Conversion of fatty acid by Bacillus sphaericus-like organisms", Curr. Microbiol., 45, pp.265~271 (2002)
  25. Schneider, S., "Biocatalyst engineering by assembly of fatty acid transport and oxidation activities for in vivo application of cytochrome P450BM-3 monooxygenase", Appl. Environ. Microbiol., 64, pp.3784~3790 (1998)
  26. Schneider, S., "Controlled regioselectivity of fatty acid oxidation by whole cells producing cytochrome P450BM-3 monooxygenase under varied dissolved oxygen concentration", Biotechnol. Bioeng., 64, pp.333~341 (1999)
  27. Sauveplane, V., "Arabidopsis thaliana CYP77A4 is the first cytochrome P450 able to catalyze the epoxidation of free fatty acids in plants", FEBS J., 276, pp.719~735 (2009)
  28. Laethem, R. M., "Epoxidation of C18 unsaturated fatty acids by cytochromes P4502C2 and P4502CAA", Drug Metab. Dispos., 24, pp.664~668 (1996)
  29. Ortiz de Montellano, P. R., "Mechanism-based probes of the topology and function of fatty acid hydroxylases", FASEB J., 6, pp.695~699 (1992)
  30. Kieser, T., "Practical Streptomyces Genetics" (2000), The John Innes Centre Foundation
  31. Sambrook, J., "Molecular cloning, a laboratory manual" (2001), Cold Spring Harbor Laboratory
  32. Sugimoto, H., "Crystal structure of CYP105A1 (P450SU-1) in complex with 1 alpha,25-dihydroxyvitamin D3", Biochemistry, 47, pp.4017~4027 (2008)
  33. Zhao, B., "Role of active site water molecules and substrate hydroxyl groups in oxygen activation by cytochrome P450 158A2: A new mechanism of proton transfer", J. Biol. Chem., 280, pp.42188~42197 (2005)
  34. Xu, L. H., "Crystal structure of cytochrome P450 105P1 from Streptomyces avermitilis: conformational flexibility and histidine ligation state", J. Bacteriol., 191, pp.1211~1219 (2009)
  35. Sippl, M. J., "Recognition of errors in three-dimensional structures of proteins", Proteins, 17, pp.355~362 (1993)
  36. Lovell, S. C., "Structure validation by Calpha geometry: phi, psi and Cbeta deviation", Proteins, 50, pp.437~450 (2003)
  37. Maiti, R., "SuperPose: a simple server for sophisticated structural superposition", Nucleic Acids Res., 32, pp.W590~W594 (2004)
  38. Hussain, H. A., "Enhanced heterologous expression of two Streptomyces griseolus cytochrome P450s and Streptomyces coelicolor ferredoxin reductase as potentially efficient hydroxylation catalysts", Appl. Environ. Microbiol., 69, pp.373~382 (2003)
  39. Omura, T., "The carbon monoxide- binding pigment of liver microsomes. I. Evidence for its hemoprotein nature", J. Biol. Chem., 239, pp.2370~2378 (1964)

관련 논문AI추천

Homology Modeling and In Vitro Analysis for Characterization of Streptomyces peucetius CYP157C4오태진, 유상철, 장종화, Hemraj Rimal한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2015 · 제25권 제9호Characterization of Two Self-Sufficient Monooxygenases, CYP102A15 and CYP102A170, as Long-Chain Fatty Acid Hydroxylases박현, 김기화, 오태진, 이우행, Hemraj Rimal한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2020 · 제30권 제5호Dual positional substrate specificity of rice allene oxide synthase-1: insight into mechanism of inhibition by type II ligand imidazole한옥수, 윤세레밭, Randeep Rakwal생화학분자생물학회 · BMB Reports2013 · 제46권 제3호Crystal Structure and Functional Characterization of a Cytochrome P450 (BaCYP106A2) from Bacillus sp. PAMC 23377박현, 김기화, 박선하, 오태진, 이준혁, 이창우, Bikash Dangi한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2017 · 제27권 제8호The N-Terminal α-Helix Domain of Pseudomonas aeruginosa Lipoxygenase Is Required for Its Soluble Expression in Escherichia coli but Not for CatalysisGuangsheng Wang, Guocheng Du, Jian Chen, Song Liu, Xiaoman Zhou, Xinyao Lu, Yue Feng한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2016 · 제26권 제10호Crystal Structure and Biochemical Analysis of a Cytochrome P450 Steroid Hydroxylase (BaCYP106A6) from Bacillus SpeciesChoi Mieyoung, Do Hackwon, Kim Ki-Hwa, Lee Chang Woo, Lee Jun Hyuck, Nam Yewon, Oh Tae-Jin, Subedi Pradeep한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2023 · 제33권 제3호

이 간행물의 다른 논문 (11건)