Delta 3,delta 2-enoyl-CoA isomerases. Characterization of the mitochondrial isoenzyme in the rat.

Palosaari, P M; Kilponen, J M; Sormunen, Raija; Hassinen, Ilmo E.; Hiltunen, J. Kalervo

doi:10.1016/s0021-9258(19)39773-x

Cited by 69 publications

(9 citation statements)

References 21 publications

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“…(2) A3,A2-Enoyl-CoA Isomerase. This enzyme activity was assayed at 37 °C as described in the literature (Palosaari et al, 1990) with some modifications. The assay mixture contained 17 ^mol of EDTA, 2 /miol NAD, 0.4 unit of /3-hydroxy-acyl CoA dehydrogenase, 0.4 unit of crotonase, and tranr-3-hexenoyl-CoA (60 nmol) in 1 mL of 0.17 M Tris buffer (pH 9).…”

Section: Methodsmentioning

confidence: 99%

Purification and Mechanism of .DELTA.3,.DELTA.5-t-2,t-4-Dienoyl-CoA Isomerase from Rat Liver

Chen¹,

Jin²,

Tserng³

1994

Biochemistry

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A new enzyme, i.e., delta 3,delta 5-t-2,t-4-dienoyl-CoA isomerase, required in the NADPH-dependent metabolic pathway of odd-numbered double bond, unsaturated fatty acids, was isolated and purified to apparent homogeneity from rat liver. In the oxidation of odd-numbered double bond, unsaturated fatty acids, stepwise beta-oxidation leads to cis-5-enoyl-CoA, which is then dehydrogenated and isomerized to delta 3,delta 5-dienoyl-CoA. delta 3,delta 5-t-2,t-4-Dienoyl-CoA isomerase converts delta 3,delta 5-dienoyl-CoA to trans-2,trans-4-dienoyl-CoA, which is a substrate for NADPH-dependent 2,4-dienoyl-CoA reductase. This enzyme was purified through Matrex gel red A, blue Sepharose, DEAE-cellulose, CM-cellulose, hydroxylapatite, and Sepharose CL6B column chromatography of an ammonium sulfate precipitated fraction (30-80%) of rat liver homogenate. A native molecular weight of 200,000 with four subunits of 55,000 each was determined. The isoelectric point was 6.5. This enzyme was located in mitochondria and was inducible by clofibrate treatment. Using delta 3,delta 5-decadienoyl-CoA, delta 3,delta 5-dodecadienoyl-CoA, and delta 3,delta 5-tetradecadienoyl-CoA as substrates, the Vmax ratio was 1:0.5:0.4 and the Km's were 10.9, 5.9, and 1.4 microM, respectively. The specific activity of purified enzyme was 7 units/mg using delta 3,delta 5-decadienoyl-CoA as substrate. The mechanism of isomerization was studied by deuterium labeling. Consistent with the deuterium labeling pattern of the products, the isomerization from trans-2,cis-5-dienoyl-CoA to trans-2,trans-4-dienoyl-CoA was a two-step process through an intermediate delta 3,delta 5-dienoyl-CoA.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Methodsmentioning

confidence: 99%

Purification and Mechanism of .DELTA.3,.DELTA.5-t-2,t-4-Dienoyl-CoA Isomerase from Rat Liver

Chen¹,

Jin²,

Tserng³

1994

Biochemistry

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“…(4) A3,A2-Enoyl-CoA Isomerase. This enzyme's activity was assayed at 37 °C as described in the literature (Palosaari et al, 1990), with some modification. The assay mixture contained 17 µ of EDTA, 2 µ of NAD, 0.4 unit of /3-hydroxyacyl-CoA dehydrogenase, 0.4 unit of crotonase, and trans-3 -hexenoyl-CoA (60 nmol) in 1 mL of 0.17 M Tris buffer (pH 9).…”

Section: Methodsmentioning

confidence: 99%

Isomerization of trans-2,.DELTA.5-Dienoyl-CoA's to .DELTA.3,.DELTA.5-Dienoyl-CoA's in the .beta.-Oxidation of .DELTA.5-Unsaturated Fatty Acids

Chen

Jin²,

Dejak³

et al. 1995

Biochemistry

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The NADPH-dependent reduction pathway for the metabolism of delta 5-unsaturated fatty acids involves the isomerization of trans-2,delta 5-dienoyl-CoA, initially formed from the dehydrogenation of delta 5-enoyl-CoA, to isomeric delta 3,delta 5-dienoyl-CoA. The latter intermediates were then isomerized to trans-2,trans-4-dienoyl-CoA, which then follows the NADPH-dependent pathway mediated by 2,4-dienoyl-CoA reductase. The isomerization from trans-2,delta 5-dienoyl-CoA to delta 3,delta 5-dienoyl-CoA is catalyzed by delta 3,delta 2-enoyl-CoA isomerase. In this investigation, we identified the stereoisomers of delta 3,delta 5-dienoates that were formed in the reaction. Starting from trans-2,cis-5-decadienoyl-CoA, the isomerization produced cis-3,cis-5- and trans-3,cis-5-decadienoates. On the other hand, trans-2,trans-5-decadienoyl-CoA yielded cis-3,trans-5- and trans-3,trans-5-decadienoates. In addition to purified rat liver delta 3,delta 2-enoyl-CoA isomerase, acyl-CoA oxidase from Arthrobacter also catalyzed the isomerization from trans-2,cis-5-dienoyl-CoA. However, this acyl-CoA oxidase could not catalyze the similar isomerization of trans-2,trans-5-dienoyl-CoA. delta 3,delta 5-t-2,t-4-Dienoyl-CoA isomerase used cis-3,cis-5-, trans-3,cis-5-, and cis-3,trans-5-dienoyl-CoA's as substrates and converted them to trans-2,trans-4-dienoyl-CoA. In contrast, trans-3,trans-5-dienoyl-CoA was not a substrate for this isomerization. Extensive purification of acyl-CoA oxidase through column chromatography could not remove or diminish the isomerization activity associated with acyl-CoA oxidase. Acyl-CoA oxidases derived from Candida and rat liver also possess isomerization activity.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…The use of bioinformatics and in silico cloning has drastically improved techniques in biological research. In silico cloning is a relatively new strategy of gene cloning that takes advantage of an existing database tosave time, expenditure, and accelerates the discovery of unknown genes (Palosaari et al 1990;Gao et al 2005). In the present study, we obtained ECI1 cDNA (GenBank: KC447361.1) from three types of pig and tissue types and found a complete coding region of 909 bp encoding 302 amino acids.…”

Section: Cloning and Snp Identificationmentioning

confidence: 99%

“…Mitochondrial enoyl-CoA isomerase is an auxiliary enzyme involved in unsaturated fatty acid oxidation. Enzyme purification from tissues of rat, bovine, and humans has been reported (Euler-Bertram and Stoffel 1990;Palosaari et al 1990;Kilponen and Hiltunen 1993). A yeast mutant devoid of ECI1 has been reported unable to grow on media containing unsaturated fatty acids, such as oleic acid (Gurvitz et al 1998).…”

Section: Introductionmentioning

confidence: 99%

Cloning, expression, and polymorphism of the ECI1 gene in various pig breeds

Chen

Zhang

et al. 2017

Journal of Integrative Agriculture

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The enzyme Δ 3 ,Δ 2 -dienoyl-CoA isomerase (ECI1) plays a crucial role in the mitochondrial β-oxidation of fatty acids with a double-bond in odd and even positions. The ECI1 gene might be a qualified candidate for studies pertaining to lipid deposition and meat quality in swine. In the present study, ECI1 cDNA of the Tibetan pig was obtained by in silico cloning and verified by PCR analysis. Single-nucleotide polymorphisms (SNPs) of ECI1 were screened by PCR-sequencing and genotypes of those SNPs were tested by PCR-restriction fragment length polymorphism (PCR-RFLP) in Diannan small-ear pigs (DSP, n=40), Tibetan pigs (TP, n=60) and Yorkshire pigs (YP, n=30). The expression levels of ECI1 were analyzed by real-time quantitative PCR and Western blotting in tissues of the liver, backfat, and longissimus dorsi (LD) muscle of DSP (n=8), TP (n=8) and YP (n=8). Single factor linear correlation analysis was applied separately for each breed to evaluate correlations between ECI1 gene expression in the LD muscle and intramuscular fat (IMF) content. We obtained an ECI1 gene length of 1 401 bp from the cDNA that contained a full coding region of 909 bp. Three novel SNPs (g.42425337G>A; g.42424666A>G; and g.42422755A>G) were detected, and only g.42424666A>G exhibited three genotypes among the three breeds. The ECI1 expression levels in the LD muscle of DSP and TP were significantly higher than that of YP (P<0.05). Moreover, TP had the highest ECI1 expression in backfat (P<0.01), and a positive correlation was observed between gene expression and IMF content. The results suggest that differences in ECI1 gene expression might be related to lipid deposition and meat quality in pig.

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Delta 3,delta 2-enoyl-CoA isomerases. Characterization of the mitochondrial isoenzyme in the rat.

Cited by 69 publications

References 21 publications

Purification and Mechanism of .DELTA.3,.DELTA.5-t-2,t-4-Dienoyl-CoA Isomerase from Rat Liver

Purification and Mechanism of .DELTA.3,.DELTA.5-t-2,t-4-Dienoyl-CoA Isomerase from Rat Liver

Isomerization of trans-2,.DELTA.5-Dienoyl-CoA's to .DELTA.3,.DELTA.5-Dienoyl-CoA's in the .beta.-Oxidation of .DELTA.5-Unsaturated Fatty Acids

Cloning, expression, and polymorphism of the ECI1 gene in various pig breeds

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