Evidence for a peroxisomal fatty acid beta-oxidation involving D-3-hydroxyacyl-CoAs. Characterization of two forms of hydro-lyase that convert D-(-)-3-hydroxyacyl-CoA into 2-trans-enoyl-CoA

Eur J Biochem. 1991 Aug 15;200(1):171-8. doi: 10.1111/j.1432-1033.1991.tb21064.x.


A novel D-(-)-3-hydroxyacyl-CoA hydro-lyase, forming 2-trans-enoyl-CoA and formerly designated as epimerase (EC, was extracted from fat-degrading cotyledons of cucumber seedlings. The enzyme, called D-3-hydroxyacyl-CoA hydro-lyase or D-specific 2-trans-enoyl-CoA hydratase, is shown to be required for the degradation of unsaturated fatty acids that contain double bonds extending from even-numbered C atoms. The D-3-hydroxyacyl-CoA hydro-lyase was exclusively localized within peroxisomes. A 10,000-fold purification by chromatography on a hydrophobic matrix, a cation exchanger, on hydroxyapatite and Mono S led to two proteins of apparent homogeneity, both exhibiting Mr of 65,000. The D-3-hydroxyacyl-CoA hydro-lyases are homodimers with slightly differing isoelectric points around pH = 9.0. They catalyze the conversion of 2-trans-enoyl-CoA into D-3-hydroxyacyl-CoA. The reverse reaction was observed but no reaction with 2-cis-enoyl-CoAs or L-3-hydroxyacyl-CoAs. 2-trans-Decenoyl-CoA was converted 10-times faster than 2-trans-butenoyl-CoA. The conversion of 4-cis-decenoyl-CoA into octenoyl-CoA was demonstrated in vitro with purified proteins with an assay mixture containing acyl-CoA oxidase, multifunctional protein, thiolase and the D-3-hydroxyacyl-CoA hydro-lyase. Comparisons of enzyme activities present in the cotyledons or isolated peroxisomes clearly show that the pathway via dienoyl-CoA reductase is much less effective than the sequence involving D-3-hydroxyacyl-CoA hydro-lyase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3-Hydroxyacyl CoA Dehydrogenases / chemistry*
  • 3-Hydroxyacyl CoA Dehydrogenases / metabolism
  • Acyl Coenzyme A / metabolism
  • Chromatography, Ion Exchange
  • Electrophoresis, Polyacrylamide Gel
  • Enoyl-CoA Hydratase / chemistry*
  • Enoyl-CoA Hydratase / metabolism
  • Fatty Acids, Unsaturated / metabolism*
  • Hydro-Lyases / chemistry*
  • Hydro-Lyases / isolation & purification
  • Isoelectric Point
  • Isomerases / chemistry*
  • Isomerases / metabolism
  • Microbodies / enzymology*
  • Molecular Weight
  • Multienzyme Complexes / chemistry*
  • Multienzyme Complexes / metabolism
  • Peroxisomal Bifunctional Enzyme
  • Plants / enzymology
  • Racemases and Epimerases / chemistry*
  • Racemases and Epimerases / metabolism
  • Seeds / enzymology
  • Stereoisomerism


  • Acyl Coenzyme A
  • Fatty Acids, Unsaturated
  • Multienzyme Complexes
  • octanoyl-coenzyme A
  • decanoyl-coenzyme A
  • 3-Hydroxyacyl CoA Dehydrogenases
  • D-3-hydroxyacyl CoA dehydratase
  • Hydro-Lyases
  • Enoyl-CoA Hydratase
  • Peroxisomal Bifunctional Enzyme
  • Isomerases
  • Racemases and Epimerases
  • 3-hydroxybutyryl-CoA epimerase