P450BM-3: Absolute configuration of the primary metabolites of palmitic acid

Gilles Truan, Malla R. Komandla, J R Falck, Julian A. Peterson

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

P450BM-3, a catalytically self-sufficient, soluble bacterial P450, contains on the same polypeptide a heme domain and a reductase domain. P450BM-3 catalyzes the oxidation of short- and long-chain, saturated and unsaturated fatty acids. The three-dimensional structure of the heme domain both in the absence and in the presence of fatty acid substrates has been determined; however, the fatty acid in the substrate-bound form is not adequately close to the heme iron to permit a prediction regarding the stereoselectivity of oxidation. In the case of long-chain fatty acids, the products can also serve as substrate and be metabolized several times. In the current study, we have determined the absolute configuration of the three primary products of palmitic acid hydroxylation (15-, 14-, and 13-OH palmitic acid). While the 15- and 14-hydroxy compounds are produced in a highly stereoselective manner (98% R, 2% S), the 13-hydroxy is a mixture of 72% R and 28% S. We have also examined the binding of these three hydroxy acids to P450BM-3 and shown that only two of them (14-OH and 13-OH palmitic acid) can bind to and be further metabolized by P450BM-3. The results indicate that in contrast to the flexibility of palmitoleic acid bound to the oxidized enzyme, palmitic acid is rigidly bound in the active site during catalytic turnover.

Original languageEnglish (US)
Pages (from-to)192-198
Number of pages7
JournalArchives of Biochemistry and Biophysics
Volume366
Issue number2
DOIs
StatePublished - Jun 15 1999

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Palmitic Acid
Metabolites
Fatty Acids
Heme
Substrates
Stereoselectivity
Oxidation
Hydroxylation
Hydroxy Acids
Unsaturated Fatty Acids
Catalytic Domain
Oxidoreductases
Iron
Peptides
Enzymes

Keywords

  • Bacillus megaterium
  • Monooxygenases
  • P450
  • Palmitic acid

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

P450BM-3 : Absolute configuration of the primary metabolites of palmitic acid. / Truan, Gilles; Komandla, Malla R.; Falck, J R; Peterson, Julian A.

In: Archives of Biochemistry and Biophysics, Vol. 366, No. 2, 15.06.1999, p. 192-198.

Research output: Contribution to journalArticle

Truan, Gilles ; Komandla, Malla R. ; Falck, J R ; Peterson, Julian A. / P450BM-3 : Absolute configuration of the primary metabolites of palmitic acid. In: Archives of Biochemistry and Biophysics. 1999 ; Vol. 366, No. 2. pp. 192-198.
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