title: Hydrogen peroxide-mediated dealkylation of 7-ethoxycoumarin by cytochrome P450 (CYP107AJ1) from Streptomyces peucetius ATCC27952.
abstract: Cytochrome P450 CYP107AJ1, which was isolated from Streptomyces peucetius and showed high homology with peroxygenases, catalyzed a dealkylation reaction with hydrogen peroxide to provide electrons, protons and oxygen, evading the requirement for a supporting redox protein. Preliminary investigation of its transcriptional level in S. peucetius showed significant expression. Homology modeling and subsequent docking with 7-ethoxycoumarin yielded a reasonable docked structure. cyp107AJ1 cloned into pET28a(+) was expressed in Escherichia coli, and soluble protein was subjected to column-chromatographic purification in order to carry out enzyme assays with 7-ethoxycoumarin. HPLC analysis of the extracted product, corresponding to its LC/MS analysis, showed the dealkylated 7-ethoxycoumarin, which was further established by subsequent GC/MS spectral analysis. We suggest that CYP107AJ1 bypassed the requirement for NAD(P)H and redox partners for generating novel analogues.
authors: Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, Sun Moon University, #100 Kalsan-ri, Tangjeong-myeon, Asansi, Chungnam 336-708, Republic of Korea.
authors: Narayan Prasad
authors: NP
authors: Niraula
authors: Bashistha Kumar
authors: BK
authors: Kanth
authors: Jae Kyung
authors: JK
authors: Sohng
authors: Tae-Jin
authors: TJ
authors: Oh
mesh_terms: D000595:Amino Acid Sequence; D001709:Biotechnology / Q000379:methods*; D003374:Coumarins / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D003640:Dealkylation / Q000187:drug effects; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D040681:Structural Homology, Protein
chemical_list: D003374:Coumarins; D011994:Recombinant Proteins; C017299:7-ethoxycoumarin; D003577:Cytochrome P-450 Enzyme System; D006861:Hydrogen Peroxide