Purification and Properties of 3-cis-2-trans-Enoyl-CoA Isomerase (Dodecenoyl-CoA Δ-Isomerase) from Rat Liver Mitochondria

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Abstract
The mitochondrial enzyme 3-cis-2-trans-enoyl-CoA isomerase [EC 5.5.3.8], responsible for the positional and geometric isomerization of .beta.,.gamma.-unsaturated fatty acyl-CoA intermediates arising during .beta.-oxidation of unsaturated long chain fatty acids, was isolated from rat liver, purified to homogeneity by a heat step and a combination of gel filtration and ion-exchange chromatographic procedures. The enzyme has a MW of 30,000, as determined by dodecylsulfate polyacrylamide gel electrophoresis. The isomerase has a strong tendency to form a dimer. It elutes from a calibrated Sephadex G-200 column with an apparent MW = 60,000. The basic isoelectric point, pI 9.0-9.2, is due to its high content in basic amino acids. The amino acid composition determined by the ninhydrin and o-phthalaldehyde detection method is presented.