Purification and characterization of fatty acid beta-oxidation enzymes from Caulobacter crescentus.
Acetoacetyl coenzyme A (acetoacetyl-CoA) thiolase, an enzyme required for short-chain fatty acid degradation, has been purified to near homogeneity from Caulobacter crescentus. The relative heat stability of this enzyme allowed it to be separated from beta-ketoacyl-CoA thiolase. The purification scheme minus the heating step also permitted the copurification of crotonase and 3-hydroxyacyl-CoA dehydrogenase. These activities are in a multienzyme complex in Escherichia coli, but a similar complex was not observed in C. crescentus. Instead, separate proteins differing in enzymatic activity were detected, analogous to the beta-oxidation enzymes that have been isolated from Clostridium acetobutylicum and from mitochondria of higher eucaryotes. In these cells, as appears to be the case with C. crescentus, the individual enzymes form multimers of identical subunits.
O'Connell
M A
MA
Department of Molecular Biology, Albert Einstein College of Medicine, Bronx, New York 10461.
Orr
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Shapiro
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D015094:3-Hydroxyacyl CoA Dehydrogenases / Q000302:isolation & purification* / Q000378:metabolism; D000101:Acetyl-CoA C-Acetyltransferase / Q000302:isolation & purification* / Q000378:metabolism; D000123:Acetyltransferases / Q000302:isolation & purification*; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004746:Enoyl-CoA Hydratase / Q000302:isolation & purification* / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism*; D006090:Gram-Negative Bacteria / Q000201:enzymology*; D006836:Hydro-Lyases / Q000302:isolation & purification*; D007700:Kinetics; D008970:Molecular Weight; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism; D010084:Oxidation-Reduction
D005227:Fatty Acids; D009829:Oleic Acids; D019301:Oleic Acid; D015094:3-Hydroxyacyl CoA Dehydrogenases; D000123:Acetyltransferases; D000101:Acetyl-CoA C-Acetyltransferase; D006836:Hydro-Lyases; D004746:Enoyl-CoA Hydratase