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Stereoselective formation of a K-region dihydrodiol from phenanthrene by Streptomyces flavovirens

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Abstract

The metabolism of phenanthrene, a polycyclic aromatic hydrocarbon (PAH), by Streptomyces flavovirens was investigated. When grown for 72 h in tryptone yeast extract broth saturated with phenanthrene, the actinomycete oxidized 21.3% of the hydrocarbon at the K-region to form trans-9,10-dihydroxy-9,10-dihydrophenanthrene (phenanthrene trans-9,10-dihydrodiol). A trace of 9-phenanthrol was also detected. Metabolites isolated by thin-layer and high performance liquid chromatography were identified by comparing chromatographic, mass spectral, and nuclear magnetic resonance properties with those of authentic compounds. Experiments using [9-14C]phenanthrene showed that the trans-9,10-dihydrodiol had 62.8% of the radioactivity found in the metabolites. Circular dichroism spectra of the phenanthrene trans-9,10-dihydrodiol indicated that the absolute configuration of the predominant enantiomer was (−)-9S,10S, the same as that of the principal enantiomer produced by mammalian enzymes. Incubation of S. flavovirens with phenanthrene is an atmosphere of 18O2, followed by gas chromatographic/mass spectral analysis of the metabolites, indicated that one atom from molecular oxygen was incorporated into each molecule of the phenanthrene trans-9,10-dihydrodiol. Cytochrome P-450 was detected in 105,000×g supernatants prepared from cell extracts of S. flavovirens. The results show that the oxidation of phenanthrene by S. flavovirens was both regio- and stereospecific.

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Abbreviations

CD:

circular dichroism

DMF:

N,N-dimethyl-formamide

GC/MS:

gas chromatography/mass spectrometry

HPLC:

high performance liquid chromatography

NMR:

nuclear magnetic resonance

ODS:

octadecylsilane

PAH:

polycyclic aromatic hydrocarbon

TLC:

thin-layer chromatography

TMS:

tetramethylsilane

UV:

ultraviolet

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Authors and Affiliations

  1. National Center for Toxicological Research, Food and Drug Administration, 72079, Jefferson, AR, USA

    John B. Sutherland, James P. Freeman, Allison L. Selby, Peter P. Fu, Dwight W. Miller & Carl E. Cerniglia

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  1. John B. Sutherland
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  2. James P. Freeman
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  3. Allison L. Selby
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  4. Peter P. Fu
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  5. Dwight W. Miller
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  6. Carl E. Cerniglia
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Sutherland, J.B., Freeman, J.P., Selby, A.L. et al. Stereoselective formation of a K-region dihydrodiol from phenanthrene by Streptomyces flavovirens . Arch. Microbiol. 154, 260–266 (1990). https://doi.org/10.1007/BF00248965

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