Fatty acid monooxygenation by P450BM-3: Product identification and proposed mechanisms for the sequential hydroxylation reactions

Sekhar S. Boddupalli, Bikash C. Pramanik, Clive A. Slaughter, Ronald W. Estabrook, Julian A. Peterson

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

The soluble P450 isolated from Bacillus megaterium (the product of the CYP102 gene) (P450BM-3) is a catalytically self-sufficient fatty acid hydroxylase which converts lauric, myristic, and palmitic acids to ω-1, ω-2, and ω-3 hydroxy analogs. The percentage distribution of the regioisomers depends on the substrate chain length. Lauric and myristic acids were preferentially metabolized to their ω-1 hydroxy counterparts while no hydroxylation occurred when capric acid was used as the substrate. Palmitic acid, when present at concentrations greater than the concentration of oxygen in the reaction medium (>250 μm), was hydroxylated to its ω-1, ω-2, and ω-3 hydroxy analogs, with the percentage distribution of the regioisomers being 21:44:35, respectively. No ω hydroxylation of any of the fatty acids was detected. When the concentration of palmitic acid was less than the concentration of oxygen in the reaction mixture, it was noted that a number of additional products were formed. Under these conditions, unlike lauric and myristic acids, it was observed that palmitic acid was first converted to its monohydroxy isomers which were subsequently metabolized to a mixture of 14-ketohexadecanoic, 15-ketohexadecanoic, 13-hydroxy-14-ketohexadecanoic, 14-hydroxy-15-ketohexadecanoic, and 13,14-dihydroxyhexadecanoic acids with a relative distribution of 8:2:40:30:20, respectively. Thus, P450BM-3 is able not only to monohydroxylate a variety of fatty acids but also to further metabolize some of these primary metabolites to secondary and tertiary products. The present paper characterizes the products formed during the sequential hydroxylation of palmitic acid and proposes reaction pathways to explain these results.

Original languageEnglish (US)
Pages (from-to)20-28
Number of pages9
JournalArchives of Biochemistry and Biophysics
Volume292
Issue number1
DOIs
StatePublished - Jan 1 1992
Externally publishedYes

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Hydroxylation
Lauric Acids
Palmitic Acid
Fatty Acids
Oxygen
Bacillus megaterium
Bacilli
Substrates
Metabolites
Mixed Function Oxygenases
Chain length
Isomers
Genes
Acids

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Fatty acid monooxygenation by P450BM-3 : Product identification and proposed mechanisms for the sequential hydroxylation reactions. / Boddupalli, Sekhar S.; Pramanik, Bikash C.; Slaughter, Clive A.; Estabrook, Ronald W.; Peterson, Julian A.

In: Archives of Biochemistry and Biophysics, Vol. 292, No. 1, 01.01.1992, p. 20-28.

Research output: Contribution to journalArticle

Boddupalli, Sekhar S. ; Pramanik, Bikash C. ; Slaughter, Clive A. ; Estabrook, Ronald W. ; Peterson, Julian A. / Fatty acid monooxygenation by P450BM-3 : Product identification and proposed mechanisms for the sequential hydroxylation reactions. In: Archives of Biochemistry and Biophysics. 1992 ; Vol. 292, No. 1. pp. 20-28.
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